research article antioxidant and wound healing activity of

Research ArticleAntioxidant and Wound Healing Activity of PolyherbalFractions of Clinacanthus nutans and Elephantopus scaber

Muhammad Shahzad Aslam1 Muhammad Syarhabil Ahmad1 Awang Soh Mamat1

Muhammad Zamharir Ahmad2 and Faridah Salam2

1School of Bioprocess Engineering Universiti Malaysia Perlis Kompleks Pusat Pengajian Jejawi 3 (KPPJ3)Kawasan Perindustrian Jejawi 02600 Arau Perlis Malaysia2Biotechnology and Nanotechnology Research Center Malaysian Agricultural Research and Development Institute43400 Serdang Selangor Malaysia

Correspondence should be addressed to Muhammad Shahzad Aslam aslammuhammadshahzadgmailcom

Received 21 May 2016 Revised 14 June 2016 Accepted 22 June 2016

Academic Editor Yoshiyuki Kimura

Copyright copy 2016 Muhammad Shahzad Aslam et al This is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

Elephantopus scaber and Clinacanthus nutans are traditionally used as wound healing herb The objective of the present study isto develop a new polyherbal formulation by comparison the herbal combination of Elephantopus scaber and Clinacanthus nutansas an in vitro antioxidant activity with their individual herbal activity followed by fractionation of polyherbal formulation forin vivo wound healing activities and identification of bioactive compounds from their active fractions Antioxidant activity wasperformed in vitro by DPPH scavenging antioxidant activity followed by in vivo wound healing activities using excision woundmodel incision wound model and burn wound model Toxicity of the fractions of the polyherbal formulation was performedby a dermal toxicity test The result showed that Elephantopus scaber crude extract on the basis of EC

50performs a much faster

action (1567 120583gmL) but with less inhibition (8766) as compared to the combination of the new polyherbal formulation ofcrude extract (30 120583gmL) The polyherbal formulation has the highest inhibition (8949) at the same dose as compared toElephantopus scaber (8766) In comparison among all crude and fractions of new polyherbal formulation it was found that theethyl acetate fraction of polyherbal formulation has the fastest activity (EC

501483 120583gmL)with inhibition (8928) Furthermore

during evaluation of wound contraction on excision and incision woundmodel ethyl acetate fraction possesses the highest activitywith (P lt 0001) and (P lt 00001) respectively During burn wound model aqueous fraction (P lt 0001) possesses the highestactivity followed by an ethyl acetate fraction (P lt 00001) LC-MS analysis discovered the presence of several flavonoid-basedcompounds that work synergistically with sesquiterpene lactone and other bioactive compounds In conclusion flavonoid increasesthe antioxidant activity that surges the rate of wound contraction and works synergistically with other bioactive compounds

1 Introduction

Everyone experiences a fair share of wounds during thecourse of their lives The impaired and aberrant woundhealing imposes a huge financial burden in the developedworld and is an insurmountable problem in the undevelopedone Many new approaches such as gene therapy and tissueengineered skin have met with limited success Conven-tional treatment of wound consists of nonsteroidal anti-inflammatory drugs (NSAIDs) antibiotics and topical corti-costeroidwhich havemany drawbacks All of themhave somenegative impact on healing [1 2] Herbal wound products

have been an area of remarkable growth to understand thedetails of the wound healing response The field of biologicwound products aims to accelerate healing by augmentingor modulating inflammatory mediators such as eicosanoidscytokines nitric oxide and various growth factors [3] Herb-herb combinations have been used in traditional AyurvedicUnani Chinese and Peruvian medicine practice for thou-sands of years yet scientific evidence of potential bioactivecompounds and their therapeutic benefits is lacking [4 5]

The selection of herbs is based upon their activity andimportance worldwide Clinacanthus nutans Lindau is used

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2016 Article ID 4685246 14 pageshttpdxdoiorg10115520164685246

2 Evidence-Based Complementary and Alternative Medicine

for the treatment of burns eczema and herpes simplex It haslong been used as a traditional medicine for the treatment ofinsect and snakebites and skin rashes [6] Payayor (Clinacan-thus nutans) is a herbal therapy popular in Southeast AsiaA small study suggests that herbal Payayor may be superiorto benzydamine for prevention of oral mucositis [7] Leavesand even the whole plant ofClinacanthus nutans possess anti-inflammatory properties [8] The plant contains lupeol 120573-sitosterol stigmasterol botulin andmyricyl alcohol [9 10] Italso contains six known C-glycosyl flavones isolated from n-BuOH and a water soluble portion of the methanolic extractof the stems and leaves such as vitexin isovitexin shafto-side isomollupentin 7-O-120573-glucopyranoside orientin andisoorientin Five sulfur-containing glucosides were isolatedfrom the n-butanol soluble portion of a methanolic extractof the stems and leaves of plant material [11] A mixtureof cerebrosides and monoacylmonogalactosylglycerol wereseparated from the leaves of Clinacanthus nutans [9] Cli-nacanthus nutans commonly known in Malaysia as BelalaiGajah identifies as areas of initial focus under Entry PointProject (EPP) by Government of Malaysia [12]

Elephantopus scaber is commonly known in Malaysiaas Tutup Bumi Hydroalcoholic extract possesses anti-inflammatory activity A higher dose of compound signif-icantly reduced carrageenan-induced pedal edema (57)and formalin-induced pedal edema in rats (58) Ethanolicextract of leaves has shown significant antiasthmatic activitywound healing and nephroprotective activities along with itsprominent antiplatelet activity Deoxyelephantopin isolatedfrom the ethanolic extract of leaves promotes significantwound healing activity by increasing cellular proliferationthe formation of granulation tissue and synthesis of collagenand by increasing the rate of wound contraction [13] Teng-Khia-U a polyherbal formulation containing Elephantopusscaber is a Taiwan traditional medicine formulated for treat-ing nephritis edema dampness chest pain and fevercoughof pneumonia and scabiesarthralgia that was caused by thewound Researchers have shown that Teng-Khia-U possessedhepatoprotective and anti-inflammatory activity [14 15]This research affords an opportunity to develop new herbalformulation using Clinacanthus nutans and Elephantopusscaber to determine the antioxidant profile in comparisonwith individual herbs The active fractions of the polyherbalformulationwill further evaluate for its in vivowoundhealingactivity and their bioactive compounds The objective ofthe study was an investigation of the in vitro antioxidantactivity of Clinacanthus nutans Elephantopus scaber andpolyherbal extract of both medicinal herbs followed by theirfractionation After the antioxidant activity fractions of poly-herbal extract were further evaluated for their wound healingactivities Furthermore the suggested idea of investigatingbioactive compounds from active polyherbal fractions willhelp to identify several classes of phytochemicals such asflavonoids that are strongly associated with antioxidant andwound healing activity

2 Material and Method

21 Plant Material The leaf sample of these plant species wascollected from the soil of Agrotech Research Centre Institute

Of Sustainable Agrotechnology University Malaysia PerlisSg ChuchuhCampus Padang Besar PerlisThe samples werebrought to the laboratory and washed under running waterto get rid of dirt They were then dried under shade for twoweeksThematerials were pulverized in an electric mixer andpreserved in labeled glass bottles that were sealed and kept inthe refrigerator for later use

22 Preparation of Plant Extract The conventional Soxhletextraction apparatus has been used consisting of a condensera Soxhlet chamber and an extraction flask The time periodfor Soxhlet extraction was 12 hours 10 gm of dried andground leaves of Clinacanthus nutans Elephantopus scaberand combination of both herbs in equal amount (1 1)was placed in a Soxhlet apparatus and extracted with 50aqueous ethanol for 16 hours The crude extract solutionsobtained were then concentrated using a vacuum rotaryevaporator (Eyela Rotavap N-1200 vacuum controller-NVC-2200 Eyela water bath OSB-2100 Eyela chiller CCA-1111Rikakikai Co Ltd Tokyo) at a temperature of 50ndash60∘C The50 ethanol extract was fractionated using different solventsnamely hexane chloroform ethyl acetate n-butanol andwater (Figure 1)The supernatant was filtered usingWhatmannumber 1 sheet pooled and concentrated using vacuumrotary evaporatorThematerials were dried in an oven at 37∘Cfor 2 days Dried powder was stored in screw-capped glassbottles and kept in a refrigerator at 4∘C until further use

23 Cream Formulation 20 gm of powder of Clinacanthusnutans and Elephantopus scaber was extracted by Soxhletextraction Filter it 150mL of a solution of the polyherbalformulation is mixed with equal amount of infused olive oilAfter then we add 120mg of honey 100mL of rest of solutionof polyherbal extract and 20 g of beeswax

24 Free Radical Scavenging Activity Determination Thestable 22-diphenyl-1-picrylhydrazyl (DPPH) was used fordetermination of free radical scavenging activity of theextracts with fewmodifications [16] Different concentrations(20ndash200120583gmL) of each crude herbal extract were addedat an equal volume to an ethanolic solution of DPPH(01mM) After 30min at room temperature the absorbancewas recorded at 517 nm using UVVisible Spectrophotometer(UV-1800 Shimadzu Japan) The experiment was repeatedfor three times Ascorbic acid was used as positive controlsThe radical scavenging activity was expressed as the inhibi-tion percentage and monitored as per the equation DPPHradical scavenging = (AC minus ASAC) times 100 AC = absorbanceof control and AS = absorbance of the sample solutionEC50

values denote the concentration of the sample whichis required to scavenge 50 of DPPH free radicals It wascalculated from inhibition curve

25 Experimental Animals Healthy Swiss albino mice ofeither sex with more than 6ndash8 weeks of age and weightaround 45ndash60 g were used The animals were kept at roomtemperature (27 plusmn 2∘C 70ndash80 humidity 12 h lightdarkcycle) acclimatize for at least 72 h prior to the procedureCommercial food pellets and water were supplied ad libitum

Evidence-Based Complementary and Alternative Medicine 3

Leaves powder

50 ethanol extract

n-Hexanefraction

Extracted with chloroform thrice

Remaining extract

Remaining extract

Remaining extract

Extracted with ethyl acetate thrice

Aqueous ethanolextract

Dissolved indistilled water

Extracted withhexane thrice

Remaining extract

Chloroform fraction

n-Butanol fraction

EtOAc fractionExtracted with

n-butanol thrice

Extracted with 50 EtOHin water using Soxhletextraction method

Figure 1 Extraction and fractionation of herb-herb combination of Clinacanthus nutans and Elephantopus scaber

Animal handling and care were carried out throughout theexperiment following international laboratory animal useand care guidelines [17]

251 Grouping and Dosing of Animals For excision modelsix groups of mice each containing four animals was usedThe first group was treated with ethyl acetate fractionThe second and third groups were treated with n-butanoland aqueous fraction respectively The fourth group wastreated with a polyherbal cream The fifth and sixth groupswere treated with control without drug and standard usingpovidone iodine respectively All administrations were per-formed topically with amaximumquantity of 250mgkgdayFor burn wound model five groups of mice containing four

mice per group were used The first group was treated withethyl acetate fraction The second and third groups weretreated with n-butanol and aqueous fraction respectivelyThe fourth and fifth groups were treated with standard usingpovidone iodine and control without drug respectively Alladministrations were performed topically with a maximumquantity of 250mgkgday For incision wound model allanimals were treated in a similar fashionwith excisionwoundmodel For acute dermal toxicity all animals were treated ina similar fashion with burn wound model

26 Excision Wound Model On a wounding day animalswere anesthetized using diethyl ether and the back hair ofthe animals was depilated by shaving About 225mm2 area


was then marked and the full thickness of the marked areawas carefully excised by using sharp sterilized scissors After24 h of wound creation the sample was applied gently oncedaily according to the respective grouping as described undergrouping and dosing section to cover the wounded areauntil complete healing was achievedWound contraction andepithelization period were monitored Wound contractionwas measured as millimeter (mm2) and percent contractionevery day until complete epithelization was achieved [18]

261 Measurement of Wound Contraction The wound heal-ing progress was evaluated by measuring wound areas usinga transparency sheet and a permanent marker The evaluatedsurface area was used to calculate the percentage of woundcontraction taking initial size of the wound (225mm2) as100 [19] as shown below

Wound contraction

= Initial wound size minusspecific day wound size

Initial wound size

times 100

(1)

262 Epithelization Time Measurement The period ofepithelization was calculated as the number of days requiredfor falling off of the dead tissue remnants without any residualraw wound [20]

27 Incision Wound Model On a wounding day animalswere anesthetized in the same manner described for excisionwound model The dorsal fur of each mouse was then shavedand 3 cm long longitudinal paravertebral incision was madethrough the skin and subcutaneous tissue The parted skinwas then sutured 1 cm apart using a surgical thread andcurved needle The continuous thread on both wound edgeswas tightened for the good closure of the wounds (Figure 2)After 24 h of wound creation (on 1st day) animals weretreated as described under grouping and dosing section witha topical formulation of the vehicle extract or standard dailyfor nine days leaving out the last group which did not receiveany of the interventions The sutures were removed on day 8after incision and tensile strength was measured on the 11thpostwounding day [21]

28 BurnWoundModel The animals were anesthetized withdiethyl ether and their dorsal surface was shaved with asterile blade The shaved area was disinfected with 70(vv) ethanol Burn wounds were created on dorsal part ofshaved rats using a metal rod (25 cm diameter) heated to80ndash85∘C and exposed for 20 s After 24 h dead tissues wereexcised using sterile surgical blade [22] Measurement ofwound contraction and epithelization time was calculatedusing excision wound model

29 Acute Dermal Toxicity For dermal toxicity a total of 10(5 females and 5 males) rats were used Animals showingnormal skin texture were housed individually in a cage andacclimatized to the laboratory condition for five days priorto the test Following acclimation around 10 of the bodysurface area fur was shaved 24 h before the study from the

0

20

40

60

80

100

(Days)

w

ound

cont

ract

ion

0 5 10 15 20

Ethyl acetaten-ButanolAqueous

CreamStandardControl (untreated)

Figure 2 wound contraction among all fractions as compared tocontrol in excision wound model

dorsal area of the trunk of the test animals A limit test doseof 2000mgkg of each fraction was applied uniformly overthe shaved area for 24 h At the end of the exposure periodthe residual test substance was removed and the animals wereobserved for development of any adverse skin reactions dailyfor 14 days [23]

210 LC-MS Analysis of Fractions The chromatography wasperformed on an Agilent 1290 Infinity LC system coupledto Agilent 6520 Accurate-Mass Q-TOF mass spectrometerwith dual ESI source using an Agilent Zorbax SB-C18 column(21 times 150mm 35 120583m) Flow rate was 05mLmin and theinjection volume was 1 120583L column temperature 25∘C andautosampler temperature 4∘C Mobile phases consisted of01 formic acid in water (A) and 01 formic acid inacetonitrile (B) The column was equilibrated for 5min priorto each analysis Separation was carried in 30min under thefollowing conditions 0min 5 B 5min 5 B 20min 100B and 25min 100 B The MS acquisition was performedin negative and positive ionization information acquisition(IDA) between mz 100 and 1000 fragmentor voltage 125Vskimmer 65V drying gas 10 Lmin gas temperature 300∘Cnebulizer 45 psig acquisition rate (spectras) 103 acquisitiontime (msspectrum) 973 and transientsspectrum 9632 Theretention time and the mass obtained for the componentswere compared to Metabolomics database

211 Chemicals 22-Diphenyl-1-picrylhydrazyl (DPPH radi-cal) (Sigma-Aldrich Co St Louis MO USA) ethyl acetateethanol toluene chloroform methanol (Fisher ScientificUK) sulfuric acid hydrochloric acid ammonia glacial aceticacid ascorbic acid (HmbG) thin layer chromatographysilica gel 60 F254 precoated plates and formic acid (MerckDarmstadt Germany)

212 Statistical Analysis Statistical analysis was performedusing GraphPad Prism version 7 for Windows (Graph-Pad Software San Diego CA USA) and Microsoft Excel


Day 0 Day 3 Day 6 Day 9 Day 12 Day 15 Day 17

Untreated

Standard

Cream

Aqueous fraction

n-Butanol fraction

Ethyl acetate fraction

Figure 3 Comparison between all fractions with respect to control in excision wound model

2013 Raw data obtained from different wound models areexpressed as mean plusmn SEM 119875 values less than 005 were con-sidered to be statistically significant The data were analyzedusingGraphPad Prism version 7 forWindows and differencesamong groups were compared by one-way ANOVA followedby Dunnettrsquos test

3 Results and Discussion31 Free Radical Scavenging Activity Determination All theresults were reported based on the dry sample after Soxhletextraction Based on the results the test conducted forevaluating antioxidant activity in Clinacanthus nutans Ele-phantopus scaber and a combination of an equal proportionof both leaves and their fraction (n-hexane chloroformethyl acetate n-butanol aqueous fractions) showed that theDPPH radical scavenging activity increased with the increasein concentration Clinacanthus nutans Elephantopus scaberand new polyherbal combination crude extract exhibitedantioxidant activity at all the concentration of test solutionswith maximum inhibition by new polyherbal formulation(8949) With the increasing concentration of leaves extract(20ndash200120583gmL) the percentage of antioxidant activity alsoincreased On the basis of the highest inhibition theformulation was further fractionated into n-hexane chloro-form ethyl acetate n-butanol and aqueous fractions Duringevaluation of fractions of new polyherbal formulation itwas found that maximum inhibition was achieved byan n-butanol fraction (8997) followed by ethyl acetate(8928) and aqueous fraction (8620) respectively EC

50

of all samples is mentioned in Table 1Data are expressed as the concentration necessary to

scavenge 50 of DPPH radical All measurements wererepeated three times

Table 1 EC50

of crude Clinacanthus nutans Elephantopus scaberClinacanthus nutans + Elephantopus scaber (polyherbal formula-tion) and their fractions

Sample EC50(120583gmL)

Clinacanthus nutans 10328Elephantopus scaber 1567Clinacanthus nutans + Elephantopus scaber 3000n-Hexane fraction 28511Chloroform fraction 24822Ethyl acetate fraction 1483n-Butanol fraction 8974Aqueous fraction 8628Ascorbic acid 820

32 Excision Wound Model Significant wound contractionwas initiated from day 6 in Group I (P lt 005) and epithe-lization was completed on day 12 (P lt 0001) followed byGroups II (P lt 0001) and III (P lt 00001) on day 12 and day 15respectively (Table 2 Figure 3) Figure 2 represents woundcontraction among all fractions as compared to control inexcision wound model

33 Incision Wound Model The tensile strength of animalstreated with the ethyl acetate fraction was significantly higher(119875 lt 00001) than the standard followed by n-butanol (119875 lt00001) and aqueous fraction (119875 lt 00001) although noapparent difference was detected with cream of polyherbalextract (119875 lt 0001) as compared to n-butanol fraction(Table 3)

34 Burn Wound Model Wound contraction is anotherparameter used to assess wound healing Significant wound


Table2Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringexcisio

nwou

ndmod

el

Days

Day

0(beforetreatment)

36

912

1517

Group

1(ethylacetate)

229plusmn14

7(000)

1695plusmn210lowast(2598)

9025plusmn17

0lowast(6058)

6150plusmn18

4lowast(7314)

1950plusmn095lowastlowast(914

8)000plusmn000

(100)

000plusmn000

(100)

Group

2(n-butanol)

2312

5plusmn330

(000)

147plusmn12

2lowastlowast(3643)

1175plusmn10

4(491

8)48plusmn12

2lowastlowast(792

4)21plusmn241lowastlowastlowast(9091)

000plusmn000

(100)

000plusmn000

(100)

Group

3(aqu

eous)

2335plusmn589

(000)


118plusmn336

(494

6)1095plusmn221lowastlowast(5310)

7425plusmn19

3(6820)

2850plusmn327lowastlowastlowast(877

9)000plusmn000

(100)

Group

4(cream

)22825plusmn423

(000)

196plusmn294

(1412

)14075plusmn306lowastlowast(3833)

88plusmn12

2(614

4)595plusmn16

5lowastlowast(7393)


2)000plusmn000

(100)

Group

5(con

trol)

2292

5plusmn245

(000)

20325plusmn249

(114

3)118plusmn313

(4852)

9375plusmn375

(591

0)845plusmn35(6314)

54plusmn408

(7644

)305plusmn386

(8669)

Group

6(stand

ard)

2312

9plusmn473

(000)

14825plusmn19

3lowastlowast(3589)

8925plusmn228lowast(614

0)44


185plusmn272lowastlowast(92)

000plusmn000

(100)

000plusmn000

(100)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou

plowast119875lt005lowastlowast119875lt0001andlowastlowastlowast119875lt00001comparedto

respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)


Counts versus acquisition time (min)

002040608

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times107 +ESI TIC Scan Frag = 1250V EtOAc-MS-Pos-20_30-5uLd

(a)

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

01234

times106


minusESI TIC Scan Frag = 1250V EtOAc-MS-Neg-20_30-5uLd

(b)

Figure 4 LC-MS report of (a) ethyl acetate MS+ and (b) ethyl acetate MSminus

0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106

+ESI TIC Scan Frag = 1250V n-BuOH-MS-Pos-20_30-5uLd

(a)

01234

minusESI TIC Scan Frag = 1250V n-BuOH-MS-Neg-20_30-5uLd


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106

(b)

Figure 5 LC-MS report of (a) n-butanol fraction MS+ and (b) n-butanol fraction MSminus

02468


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106 +ESI TIC Scan Frag = 1250V Aq-MS-Pos-20_30-5uLd

(a)

0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106 minusESI TIC Scan Frag = 1250V Aq-MS-Neg-20_30-5uLd

(b)

Figure 6 LC-MS report of (a) aqueous fraction MS+ and (b) aqueous fraction MSminus

Table 3 Breaking strength of ethyl acetate n-butanol and aqueousfraction of Clinacanthus nutans + Elephantopus scaber (polyherbalformulation)

Groups Breaking strength (g)Ethyl acetate 73075 plusmn 756lowastlowastlowast

n-Butanol 6945 plusmn 634lowastlowastlowast

Aqueous 624 plusmn 594lowastlowastlowast

Cream 66325 plusmn 537lowastlowast

Standard 40875 plusmn 426lowastlowast

Control 3375 plusmn 661Values are mean plusmn SEM of 4 rats in each group lowastlowast119875 lt 0001 and lowastlowastlowast119875 lt00001 compared to respective day control group (statistical analysis wasdone by one-way analysis of variance followed by Dunnettrsquos test for multiplecomparisons)

contraction was initiated from day 12 in Group III (119875 lt0001) followed by Groups I (P lt 00001) and II (P lt 005)on day 15 (Table 4)

35 Acute Dermal Toxicity In acute dermal toxicity studiesthe rats of either sex were given ethyl acetate n-butanoland aqueous fractions with doses (2000mgKgday bodyweight) for 10 days The dose did not produce any signs ofinflammation The animals were physically active and wereconsuming food andwater in a regular wayWe did not noticeany abnormal behavior

36 LC-MS Profile of Active Fractions Themolecular weightof the active compound was determined with electrosprayionization mass spectrometry (ESI-MS) The ESI-MS spec-trum of the active compound is shown in spectrophotome-ter detection (LC-MS) provided by fragmentation patternAnalysis can use ESI positive [M+H]+ ion and negativecharges [MminusH]minus ion Figures 4 5 and 6 represent thefragmentation pattern of the bioactive compounds detectedfrom isolated ethyl acetate n-butanol and aqueous fractionrespectively The LC-MSMS results showed spectral data ofpossible flavonoids identified in all bioactive fractions (ethylacetate n-butanol fraction) of new polyherbal formulation


Table4Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringbu

rnwou

ndmod

elDays

Day

0(beforetreatment)

36

912

1517

19Group

1(ethylacetate)

499plusmn353

(000)

2917






000plusmn000

(000)

000plusmn000

(000)

Group

2(n-butanol)

47675plusmn92

5(000)

3717




9)13875plusmn74lowast(7089)


6)000plusmn000

(000)

000plusmn000

(000)

Group

3(aqu

eous)

4897

5plusmn18

4(000)

3112


)1772


815plusmn15

5lowastlowastlowast(8335)

485plusmn15

5lowastlowast(9009)

000plusmn000

(000)

000plusmn000

(000)

000plusmn000

(000)

Group

4(stand

ard)

492plusmn402

(000)


3175plusmn337

(3546




000plusmn000

(000)

000plusmn000

(000)

Group

5(con

trol)

4965plusmn560

(000)

42425plusmn840

(1455)

324plusmn74

2(3474)

3235plusmn75

(3484)

2072

5plusmn592

(5825)

1195plusmn613

(7593)

565plusmn575

(8862)

2675plusmn550

(9461)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)


Table5Listof

flavono

idstentativ

elyidentifi

edin

ethylacetatefractio

ns

Characteris

tics

Ethylacetatefractio

nMS+M

SminusSequ

ence

12

34

56

78

910

1112

1314

15119905119877

906

9333

9598

9694

10354

10602

11984

12816

12953

15021

11308

11316

1146

11857

14089

[MminusH](119898119911)

5801439

5641492

4481018

5341379

5161279

4020963

28604

853300746

3621373

3581422

5461368

4321426

4461597

6361493

3901325

Error(pp

m)

minus19

1minus23

minus27

minus10

5minus216

minus305

minus249

minus19

minus19

5minus15

910

3minus13

1minus459

minus226

minus273

Molecular

form

ula

C 26H28O15

C 26H28O14

C 21H20O11

C 25H26O13

C 25H24O12

C 20H18O9

C 15H10O6

C 17H14O7

C 19H22O7

C 20H22O6

C 26H26O13

C 22H24O9

C 23H26O9

C 32H28O14

C 20H22O8

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KL

MN

O

Aisoorientin

210158401015840-O

-apiofuranoside1B

vitexin210158401015840-O

-xyloside2

Cscutellarein6-glucoside3

D6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4E

apigenin

7-(210158401015840310158401015840-diacetylglucosid

e)5

Fcerarvensin

6G

572101584031015840-te

trahydroxyflavone

7H5

2101584041015840-tr

ihydroxy-68-dim

etho

xyflavone

(rehderia

ninI)

8Imachaerol

B9

Jbrosim

acutin

B10K

isorham

netin

3-[610158401015840-(2-(E)-bu

teno

yl)-glucoside]

11L

3567310158404101584051015840-

heptam

etho

xyflavone

12M

5-hydroxy-78-dim

etho

xyflavano

ne5-rham

nosid

e13Nkaempferol3-(410158401015840-acetyl-610158401015840-p-cou

marylglucoside)

14O

apigeniflavan5-O-xyloside1

5


O

O

HO

OHO

HO

HO

OH

OH

OH

OH

OH

OO

1

O

O

OOO

2

O

O

O

3

O

O

O

O

4

OH

O

OOH

OHO

OO

OO

O 5

OH

O

O

OHO

HO

OHHOHO

6

O

OOH

HO

7

OH

OH O

OOH

HO

8

OHO

O

HO

HO

9

OHO

HO

OH

OH

10

HO

OH

OHOH

OH

OH

OH

O

O

O

O11

HO

HO

HOHO

HO

HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

CH2OH

OH

OH

OH OHOH

OH

OH

OHCH2OH

CH2

OCH3

OCH3

OCH3

OCH3

OCH3

CH3

CH3

CH3

OH

H3CO

H3C

H3CO

CH2

CH3

O

Figure 7 Continued


O

O

12

O

O

HO

O

OHHO

HO

13

O

OH

HO

O

HO

HOOH

14

O

OH

HO

15

OH O

OO

HOOH

OH

OO

O

O

O

OO

O

O

O

HO

16

O OHO

OH

OH

OH

O O

CO

O

HO

OH

OH

O

OO

HO

HO

HOHO

HO

HO

O

OH

HO

17

HO

OH

OH

O

O

OOH

HOHO

HO

OHO

HO

HO

HO

O

O

18

O

OH

OH

OH

OHO

HO

HO

OH

19

O

HO

HO

OH

H3CO

H3CO

OCH3

OCH3

OCH3

OCH3

OCH3

H3C

OCH3

OCH3

H3C

CH2

CH2

CH3

CH3

CH3

CH3

CH2

2H

H3C

O

OO

O

Figure 7 Continued


OO

OH

OH OH

O

20

OHO

HO

HO

HO

O

OH

O

21

OHOOC

HO

HO

HO

OH

HO

O

O22

OH

O O

O

OO

O

23

HO

HO

HO

HO

HO

HO

OH

OH

OHOH

OH

HOOC

OH

H2C O

O

Figure 7 List of flavonoids identified from all fractions

Table 6 List of flavonoids tentatively identified in n-butanol fractions

Characteristics n-Butanol fraction MS+MSminus

Sequence 1 2 3 4 5 6 7119905119877

9058 9468 9699 10363 8735 8928 9328[MminusH] (119898119911) 5801439 448101 5341386 5161279 7061847 6101533 5641487Error (ppm) minus186 minus103 minus237 minus222 nf 008 minus135Molecular formula C

26H28O15

C21H20O11

C25H26O13

C25H24O12

C32H34O18

C27H30O16

C26H28O14

Proposed compound A B C D E F GA isoorientin 7-O-rhamnoside 19 B scutellarein 6-glucoside 3 C = 68-Di-C-120573-D-arabinopyranosylapigenin 4 D apigenin 7-(210158401015840310158401015840-diacetylglucoside) 5E kaempferol 3-[210158401015840101584031015840101584010158405101584010158401015840-triacetyl-120572-L-arabinofuranosyl-(1 rarr 6)-glucoside 16 F 6-hydroxyluteolin 7-rutinoside 17 G vitexin 210158401015840-O-xyloside 2 119905119877 retention time nf not found

from leaves of Clinacanthus nutans and Elephantopus scaberdetected with MS in negative and positive modes The spec-tral data from the peaks were identical and the identificationwas based on the LC-MSMS data and comparison with theliteratureTheoverview is shown in the chromatographic dataof ethyl acetate the n-butanol and aqueous fraction which issummarized in Tables 5 6 and 7 respectively A maximumnumber of flavonoids were found in ethyl acetate fractionfollowed by an aqueous fraction and an n-butanol fractionFlavonoids play an important role in wound healing [24ndash26]There is another important bioactive compound also presentin all fractions Among them sesquiterpene lactone and phe-nolic compounds are also present that showed the synergisticeffect Although no report was found on the synergistic effectof flavonoids and sesquiterpene lactone further investigationis going on to understand the relationship between flavonoidsand sesquiterpene lactone List of possible flavonoids is alsomentioned along with their structure in Figure 7

4 ConclusionPolyherbal formulation is a common practice in herbalismAn effort has been made to develop a new polyherbalformulation in the treatment of the wound This is the firstdocument that fractionates polyherbal extract and identifiesthe possible bioactive compound This paper has establisheda correlation between wound healing activity and possiblebioactive compounds This paper indicates that flavonoidsmay perform an important role in the healing of the woundA further experiment is required to isolate the possiblebioactive compound from ethyl acetate and an aqueousfraction of polyherbal extract and to evaluate the isolatedcompounds on different wound model

Competing InterestsThe authors declare no potential competing interests withrespect to the research authorship andor publication of thispaper


Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O

-p-hydroxybenzoagte1

8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol

341015840-dixyloside22J5

6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments

This research was supported by the Prototype Research GrantScheme (PRGS Reference no PRGS9013-00016) awardedby the Ministry of Higher Education (MOHE) MalaysiaUniversiti Malaysia Perlis (UniMAP) Ministry of HigherEducation Malaysia [Malaysian International ScholarshipReference no KPTB600-183 JLD 6 (31)] Research Col-laborative Effort from Malaysian Agricultural Research andDevelopment Institute Serdang Malaysia and Jeffrey CheahSchool of Medicine amp Health Sciences Monash UniversityMalaysia Jalan Lagoon Selatan Bandar Sunway SelangorDarul Ehsan Malaysia (Reference no 25912-1)

References

[1] D C LeBert and A Huttenlocher ldquoInflammation and woundrepairrdquo Seminars in Immunology vol 26 no 4 pp 315ndash3202014

[2] T K Biswas S Pandit and S Chakrabarti ldquoTest on Indian andPeruvian medicinal plants for wound healingrdquo in Evaluation ofHerbal Medicinal Products P K Mukherjee Ed pp 228ndash241Pharmaceutical Press London UK 2009

[3] P S Murphy and G R D Evans ldquoAdvances in wound healinga review of current wound healing productsrdquo Plastic SurgeryInternational vol 2012 Article ID 190436 8 pages 2012

[4] S G Bhope D H Nagore V V Kuber P K Gupta andM J Patil ldquoDesign and development of a stable polyherbalformulation based on the results of compatibility studiesrdquoPharmacognosy Research vol 3 no 2 pp 122ndash129 2011

[5] C-T Che Z J Wang M S S Chow and C W K Lam ldquoHerb-herb combination for therapeutic enhancement and advance-ment theory practice and future perspectivesrdquo Molecules vol18 no 5 pp 5125ndash5141 2013

[6] V Baimai and Y W Brockelman ldquoBiodiversity research andtraining program inThailandrdquo Pure and Applied Chemistry vol70 no 11 pp 2073ndash2078 1998

[7] P Putwatana P Sanmanowong O Ladawal J Tiraporn PSupaporn and N Ladawan ldquoRelief of radiation-induced oralmucositis in head and neck cancerrdquo Cancer Nursing vol 32 no1 pp 82ndash87 2009

[8] P Wanikiat A Panthong P Sujayanon C Yoosook A GRossi and V Reutrakul ldquoThe anti-inflammatory effects and theinhibition of neutrophil responsiveness byBarleria lupulina andClinacanthus nutans extractsrdquo Journal of Ethnopharmacologyvol 116 no 2 pp 234ndash244 2008

[9] P Tuntiwachwuttikul Y Pootaeng-On P Phansa and W CTaylor ldquoCerebrosides and a monoacylmonogalactosylglycerolfrom Clinacanthus nutansrdquo Chemical amp Pharmaceutical Bul-letin vol 52 no 1 pp 27ndash32 2004

[10] M S Aslam M S Ahmad and A S Mamat ldquoReview onphytochemical constituents and pharmacological activities ofClinacanthus nutansrdquo International Journal of Pharmacy andPharmaceutical Science vol 7 no 2 pp 30ndash33 2015

[11] K-I Teshima T Kaneko K Ohtani et al ldquoSulfur-containingglucosides from Clinacanthus nutansrdquo Phytochemistry vol 48no 5 pp 831ndash835 1998

[12] J P Menter ldquoETP economic transformation programmerdquoAnnual Report 2013

[13] V Krishna K L Mankani B K Manjunatha S M Vidya YN Manohara and S D J Singh ldquoWound healing activity of the

leaf extracts and deoxyelephantopin isolated fromElephantopusscaber Linnrdquo Indian Journal of Pharmacology vol 37 no 4 pp238ndash242 2005

[14] C-C Lin C-C Tsai and M-H Yen ldquoThe evaluation ofhepatoprotective effects of Taiwan folk medicine Teng-Khia-UrdquoJournal of Ethnopharmacology vol 45 no 2 pp 113ndash123 1995

[15] C-C Tsai and C-C Lin ldquoAnti-inflammatory effects of Taiwanfolk medicine lsquoTeng-Khia-Ursquo on carrageenan- and adjuvant-induced paw edema in ratsrdquo Journal of Ethnopharmacology vol64 no 1 pp 85ndash89 1998

[16] A Guerrini G Sacchetti A Grandini M Asanza and LScalvenzi ldquoCytotoxic effect and TLC bioautography-guidedapproach to detect health properties of amazonian hedyosmumSprucei essential oilrdquoEvidence-BasedComplementary andAlter-native Medicine vol 2016 Article ID 1638342 8 pages 2016

[17] Institute for Laboratory Animal Research (ILAR) Guide for theCare and Use of Laboratory Animals National Academy PressWashington DC USA 1996

[18] DDKokane R YMoreM BKaleMNNehete P CMehen-dale and C H Gadgoli ldquoEvaluation of wound healing activityof root of Mimosa pudicardquo Journal of Ethnopharmacology vol124 no 2 pp 311ndash315 2009

[19] Y Shivhare P K Singour U K Patil and R S Pawar ldquoWoundhealing potential of methanolic extract of Trichosanthes dioicaRoxb (fruits) in ratsrdquo Journal of Ethnopharmacology vol 127 no3 pp 614ndash619 2010

[20] J J Wesley A J M Christina N Chidambaranathan and KRavikumar ldquoWound healing activity of the leaves of Tribulusterrestris (linn) aqueous extract in ratsrdquo Journal of PharmacyResearch vol 2 no 5 pp 841ndash843 2009

[21] SMurthyM K Gautam S Goel V Purohit H Sharma and RK Goel ldquoEvaluation of in vivowound healing activity of Bacopamonniera on different wound model in ratsrdquo BioMed ResearchInternational vol 2013 Article ID 972028 9 pages 2013

[22] K S Priya A Gnanamani N Radhakrishnan and M BabuldquoHealing potential of Datura alba on burn wounds in albinoratsrdquo Journal of Ethnopharmacology vol 83 no 3 pp 193ndash1992002

[23] E Mulisa K Asres and E Engidawork ldquoEvaluation of woundhealing and anti-inflammatory activity of the rhizomes ofRumex abyssinicus J (Polygonaceae) in micerdquo BMC Comple-mentary and AlternativeMedicine vol 15 no 1 article 341 2015

[24] S Lodhi and A K Singhai ldquoWound healing effect of flavonoidrich fraction and luteolin isolated from Martynia annua Linnon streptozotocin induced diabetic ratsrdquo Asian Pacific Journalof Tropical Medicine vol 6 no 4 pp 253ndash259 2013

[25] A Muralidhar K S Babu T R Sankar P Reddanna and JLatha ldquoWound healing activity of flavonoid fraction isolatedfrom the stem bark of Buteamonosperma (lam) in albinowistarratsrdquo European Journal of Experimental Biology vol 3 no 6 pp1ndash6 2013

[26] S Ambiga R Narayanan D Gowri D Sukumar and SMadhavan ldquoEvaluation of wound healing activity of flavonoidsfrom Ipomoea carnea jacqrdquo Ancient Science of Life vol 26 no3 p 45 2007

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


for the treatment of burns eczema and herpes simplex It haslong been used as a traditional medicine for the treatment ofinsect and snakebites and skin rashes [6] Payayor (Clinacan-thus nutans) is a herbal therapy popular in Southeast AsiaA small study suggests that herbal Payayor may be superiorto benzydamine for prevention of oral mucositis [7] Leavesand even the whole plant ofClinacanthus nutans possess anti-inflammatory properties [8] The plant contains lupeol 120573-sitosterol stigmasterol botulin andmyricyl alcohol [9 10] Italso contains six known C-glycosyl flavones isolated from n-BuOH and a water soluble portion of the methanolic extractof the stems and leaves such as vitexin isovitexin shafto-side isomollupentin 7-O-120573-glucopyranoside orientin andisoorientin Five sulfur-containing glucosides were isolatedfrom the n-butanol soluble portion of a methanolic extractof the stems and leaves of plant material [11] A mixtureof cerebrosides and monoacylmonogalactosylglycerol wereseparated from the leaves of Clinacanthus nutans [9] Cli-nacanthus nutans commonly known in Malaysia as BelalaiGajah identifies as areas of initial focus under Entry PointProject (EPP) by Government of Malaysia [12]

Elephantopus scaber is commonly known in Malaysiaas Tutup Bumi Hydroalcoholic extract possesses anti-inflammatory activity A higher dose of compound signif-icantly reduced carrageenan-induced pedal edema (57)and formalin-induced pedal edema in rats (58) Ethanolicextract of leaves has shown significant antiasthmatic activitywound healing and nephroprotective activities along with itsprominent antiplatelet activity Deoxyelephantopin isolatedfrom the ethanolic extract of leaves promotes significantwound healing activity by increasing cellular proliferationthe formation of granulation tissue and synthesis of collagenand by increasing the rate of wound contraction [13] Teng-Khia-U a polyherbal formulation containing Elephantopusscaber is a Taiwan traditional medicine formulated for treat-ing nephritis edema dampness chest pain and fevercoughof pneumonia and scabiesarthralgia that was caused by thewound Researchers have shown that Teng-Khia-U possessedhepatoprotective and anti-inflammatory activity [14 15]This research affords an opportunity to develop new herbalformulation using Clinacanthus nutans and Elephantopusscaber to determine the antioxidant profile in comparisonwith individual herbs The active fractions of the polyherbalformulationwill further evaluate for its in vivowoundhealingactivity and their bioactive compounds The objective ofthe study was an investigation of the in vitro antioxidantactivity of Clinacanthus nutans Elephantopus scaber andpolyherbal extract of both medicinal herbs followed by theirfractionation After the antioxidant activity fractions of poly-herbal extract were further evaluated for their wound healingactivities Furthermore the suggested idea of investigatingbioactive compounds from active polyherbal fractions willhelp to identify several classes of phytochemicals such asflavonoids that are strongly associated with antioxidant andwound healing activity

2 Material and Method

21 Plant Material The leaf sample of these plant species wascollected from the soil of Agrotech Research Centre Institute

Of Sustainable Agrotechnology University Malaysia PerlisSg ChuchuhCampus Padang Besar PerlisThe samples werebrought to the laboratory and washed under running waterto get rid of dirt They were then dried under shade for twoweeksThematerials were pulverized in an electric mixer andpreserved in labeled glass bottles that were sealed and kept inthe refrigerator for later use

22 Preparation of Plant Extract The conventional Soxhletextraction apparatus has been used consisting of a condensera Soxhlet chamber and an extraction flask The time periodfor Soxhlet extraction was 12 hours 10 gm of dried andground leaves of Clinacanthus nutans Elephantopus scaberand combination of both herbs in equal amount (1 1)was placed in a Soxhlet apparatus and extracted with 50aqueous ethanol for 16 hours The crude extract solutionsobtained were then concentrated using a vacuum rotaryevaporator (Eyela Rotavap N-1200 vacuum controller-NVC-2200 Eyela water bath OSB-2100 Eyela chiller CCA-1111Rikakikai Co Ltd Tokyo) at a temperature of 50ndash60∘C The50 ethanol extract was fractionated using different solventsnamely hexane chloroform ethyl acetate n-butanol andwater (Figure 1)The supernatant was filtered usingWhatmannumber 1 sheet pooled and concentrated using vacuumrotary evaporatorThematerials were dried in an oven at 37∘Cfor 2 days Dried powder was stored in screw-capped glassbottles and kept in a refrigerator at 4∘C until further use

23 Cream Formulation 20 gm of powder of Clinacanthusnutans and Elephantopus scaber was extracted by Soxhletextraction Filter it 150mL of a solution of the polyherbalformulation is mixed with equal amount of infused olive oilAfter then we add 120mg of honey 100mL of rest of solutionof polyherbal extract and 20 g of beeswax

24 Free Radical Scavenging Activity Determination Thestable 22-diphenyl-1-picrylhydrazyl (DPPH) was used fordetermination of free radical scavenging activity of theextracts with fewmodifications [16] Different concentrations(20ndash200120583gmL) of each crude herbal extract were addedat an equal volume to an ethanolic solution of DPPH(01mM) After 30min at room temperature the absorbancewas recorded at 517 nm using UVVisible Spectrophotometer(UV-1800 Shimadzu Japan) The experiment was repeatedfor three times Ascorbic acid was used as positive controlsThe radical scavenging activity was expressed as the inhibi-tion percentage and monitored as per the equation DPPHradical scavenging = (AC minus ASAC) times 100 AC = absorbanceof control and AS = absorbance of the sample solutionEC50

values denote the concentration of the sample whichis required to scavenge 50 of DPPH free radicals It wascalculated from inhibition curve

25 Experimental Animals Healthy Swiss albino mice ofeither sex with more than 6ndash8 weeks of age and weightaround 45ndash60 g were used The animals were kept at roomtemperature (27 plusmn 2∘C 70ndash80 humidity 12 h lightdarkcycle) acclimatize for at least 72 h prior to the procedureCommercial food pellets and water were supplied ad libitum


Leaves powder

50 ethanol extract

n-Hexanefraction


Remaining extract

Remaining extract

Remaining extract





Remaining extract

Chloroform fraction

n-Butanol fraction


n-butanol thrice










Wound contraction


Initial wound size

times 100

(1)





0

20

40

60

80

100

(Days)

w

ound

cont

ract

ion

0 5 10 15 20










Untreated

Standard

Cream

Aqueous fraction

n-Butanol fraction





50



Table 1 EC50








Table2Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringexcisio

nwou

ndmod

el

Days

Day

0(beforetreatment)

36

912

1517

Group

1(ethylacetate)

229plusmn14

7(000)


9025plusmn17

0lowast(6058)

6150plusmn18

4lowast(7314)


8)000plusmn000

(100)

000plusmn000

(100)

Group

2(n-butanol)

2312

5plusmn330

(000)

147plusmn12

2lowastlowast(3643)

1175plusmn10

4(491

8)48plusmn12

2lowastlowast(792


000plusmn000

(100)

000plusmn000

(100)

Group

3(aqu

eous)

2335plusmn589

(000)


118plusmn336

(494


7425plusmn19

3(6820)


9)000plusmn000

(100)

Group

4(cream

)22825plusmn423

(000)

196plusmn294

(1412


88plusmn12

2(614

4)595plusmn16

5lowastlowast(7393)


2)000plusmn000

(100)

Group

5(con

trol)

2292

5plusmn245

(000)

20325plusmn249

(114

3)118plusmn313

(4852)

9375plusmn375

(591

0)845plusmn35(6314)

54plusmn408

(7644

)305plusmn386

(8669)

Group

6(stand

ard)

2312

9plusmn473

(000)

14825plusmn19

3lowastlowast(3589)


0)44



000plusmn000

(100)

000plusmn000

(100)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)



002040608

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(a)

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

01234

times106



(b)


0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106


(a)

01234



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106

(b)


02468


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(a)

0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(b)













Table4Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringbu

rnwou

ndmod

elDays

Day

0(beforetreatment)

36

912

1517

19Group

1(ethylacetate)

499plusmn353

(000)

2917






000plusmn000

(000)

000plusmn000

(000)

Group

2(n-butanol)

47675plusmn92

5(000)

3717






6)000plusmn000

(000)

000plusmn000

(000)

Group

3(aqu

eous)

4897

5plusmn18

4(000)

3112


)1772


815plusmn15


485plusmn15

5lowastlowast(9009)

000plusmn000

(000)

000plusmn000

(000)

000plusmn000

(000)

Group

4(stand

ard)

492plusmn402

(000)


3175plusmn337

(3546




000plusmn000

(000)

000plusmn000

(000)

Group

5(con

trol)

4965plusmn560

(000)

42425plusmn840

(1455)

324plusmn74

2(3474)

3235plusmn75

(3484)

2072

5plusmn592

(5825)

1195plusmn613

(7593)

565plusmn575

(8862)

2675plusmn550

(9461)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)


Table5Listof

flavono

idstentativ

elyidentifi

edin

ethylacetatefractio

ns

Characteris

tics

Ethylacetatefractio

nMS+M

SminusSequ

ence

12

34

56

78

910

1112

1314

15119905119877

906

9333

9598

9694

10354

10602

11984

12816

12953

15021

11308

11316

1146

11857

14089

[MminusH](119898119911)

5801439

5641492

4481018

5341379

5161279

4020963

28604

853300746

3621373

3581422

5461368

4321426

4461597

6361493

3901325

Error(pp

m)

minus19

1minus23

minus27

minus10

5minus216

minus305

minus249

minus19

minus19

5minus15

910

3minus13

1minus459

minus226

minus273

Molecular

form

ula

C 26H28O15

C 26H28O14

C 21H20O11

C 25H26O13

C 25H24O12

C 20H18O9

C 15H10O6

C 17H14O7

C 19H22O7

C 20H22O6

C 26H26O13

C 22H24O9

C 23H26O9

C 32H28O14

C 20H22O8

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KL

MN

O

Aisoorientin

210158401015840-O

-apiofuranoside1B

vitexin210158401015840-O

-xyloside2


D6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4E

apigenin


e)5

Fcerarvensin

6G

572101584031015840-te

trahydroxyflavone

7H5

2101584041015840-tr

ihydroxy-68-dim

etho

xyflavone

(rehderia

ninI)

8Imachaerol

B9

Jbrosim

acutin

B10K

isorham

netin

3-[610158401015840-(2-(E)-bu

teno

yl)-glucoside]

11L

3567310158404101584051015840-

heptam

etho

xyflavone

12M

5-hydroxy-78-dim

etho

xyflavano

ne5-rham

nosid


marylglucoside)

14O


5


O

O

HO

OHO

HO

HO

OH

OH

OH

OH

OH

OO

1

O

O

OOO

2

O

O

O

3

O

O

O

O

4

OH

O

OOH

OHO

OO

OO

O 5

OH

O

O

OHO

HO

OHHOHO

6

O

OOH

HO

7

OH

OH O

OOH

HO

8

OHO

O

HO

HO

9

OHO

HO

OH

OH

10

HO

OH

OHOH

OH

OH

OH

O

O

O

O11

HO

HO

HOHO

HO

HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

CH2OH

OH

OH

OH OHOH

OH

OH

OHCH2OH

CH2

OCH3

OCH3

OCH3

OCH3

OCH3

CH3

CH3

CH3

OH

H3CO

H3C

H3CO

CH2

CH3

O

Figure 7 Continued


O

O

12

O

O

HO

O

OHHO

HO

13

O

OH

HO

O

HO

HOOH

14

O

OH

HO

15

OH O

OO

HOOH

OH

OO

O

O

O

OO

O

O

O

HO

16

O OHO

OH

OH

OH

O O

CO

O

HO

OH

OH

O

OO

HO

HO

HOHO

HO

HO

O

OH

HO

17

HO

OH

OH

O

O

OOH

HOHO

HO

OHO

HO

HO

HO

O

O

18

O

OH

OH

OH

OHO

HO

HO

OH

19

O

HO

HO

OH

H3CO

H3CO

OCH3

OCH3

OCH3

OCH3

OCH3

H3C

OCH3

OCH3

H3C

CH2

CH2

CH3

CH3

CH3

CH3

CH2

2H

H3C

O

OO

O

Figure 7 Continued


OO

OH

OH OH

O

20

OHO

HO

HO

HO

O

OH

O

21

OHOOC

HO

HO

HO

OH

HO

O

O22

OH

O O

O

OO

O

23

HO

HO

HO

HO

HO

HO

OH

OH

OHOH

OH

HOOC

OH

H2C O

O




Sequence 1 2 3 4 5 6 7119905119877


26H28O15

C21H20O11

C25H26O13

C25H24O12

C32H34O18

C27H30O16

C26H28O14






Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O


8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol


6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Leaves powder

50 ethanol extract

n-Hexanefraction


Remaining extract

Remaining extract

Remaining extract





Remaining extract

Chloroform fraction

n-Butanol fraction


n-butanol thrice










Wound contraction


Initial wound size

times 100

(1)





0

20

40

60

80

100

(Days)

w

ound

cont

ract

ion

0 5 10 15 20










Untreated

Standard

Cream

Aqueous fraction

n-Butanol fraction





50



Table 1 EC50








Table2Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringexcisio

nwou

ndmod

el

Days

Day

0(beforetreatment)

36

912

1517

Group

1(ethylacetate)

229plusmn14

7(000)


9025plusmn17

0lowast(6058)

6150plusmn18

4lowast(7314)


8)000plusmn000

(100)

000plusmn000

(100)

Group

2(n-butanol)

2312

5plusmn330

(000)

147plusmn12

2lowastlowast(3643)

1175plusmn10

4(491

8)48plusmn12

2lowastlowast(792


000plusmn000

(100)

000plusmn000

(100)

Group

3(aqu

eous)

2335plusmn589

(000)


118plusmn336

(494


7425plusmn19

3(6820)


9)000plusmn000

(100)

Group

4(cream

)22825plusmn423

(000)

196plusmn294

(1412


88plusmn12

2(614

4)595plusmn16

5lowastlowast(7393)


2)000plusmn000

(100)

Group

5(con

trol)

2292

5plusmn245

(000)

20325plusmn249

(114

3)118plusmn313

(4852)

9375plusmn375

(591

0)845plusmn35(6314)

54plusmn408

(7644

)305plusmn386

(8669)

Group

6(stand

ard)

2312

9plusmn473

(000)

14825plusmn19

3lowastlowast(3589)


0)44



000plusmn000

(100)

000plusmn000

(100)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)



002040608

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(a)

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

01234

times106



(b)


0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106


(a)

01234



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106

(b)


02468


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(a)

0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(b)













Table4Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringbu

rnwou

ndmod

elDays

Day

0(beforetreatment)

36

912

1517

19Group

1(ethylacetate)

499plusmn353

(000)

2917






000plusmn000

(000)

000plusmn000

(000)

Group

2(n-butanol)

47675plusmn92

5(000)

3717






6)000plusmn000

(000)

000plusmn000

(000)

Group

3(aqu

eous)

4897

5plusmn18

4(000)

3112


)1772


815plusmn15


485plusmn15

5lowastlowast(9009)

000plusmn000

(000)

000plusmn000

(000)

000plusmn000

(000)

Group

4(stand

ard)

492plusmn402

(000)


3175plusmn337

(3546




000plusmn000

(000)

000plusmn000

(000)

Group

5(con

trol)

4965plusmn560

(000)

42425plusmn840

(1455)

324plusmn74

2(3474)

3235plusmn75

(3484)

2072

5plusmn592

(5825)

1195plusmn613

(7593)

565plusmn575

(8862)

2675plusmn550

(9461)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)


Table5Listof

flavono

idstentativ

elyidentifi

edin

ethylacetatefractio

ns

Characteris

tics

Ethylacetatefractio

nMS+M

SminusSequ

ence

12

34

56

78

910

1112

1314

15119905119877

906

9333

9598

9694

10354

10602

11984

12816

12953

15021

11308

11316

1146

11857

14089

[MminusH](119898119911)

5801439

5641492

4481018

5341379

5161279

4020963

28604

853300746

3621373

3581422

5461368

4321426

4461597

6361493

3901325

Error(pp

m)

minus19

1minus23

minus27

minus10

5minus216

minus305

minus249

minus19

minus19

5minus15

910

3minus13

1minus459

minus226

minus273

Molecular

form

ula

C 26H28O15

C 26H28O14

C 21H20O11

C 25H26O13

C 25H24O12

C 20H18O9

C 15H10O6

C 17H14O7

C 19H22O7

C 20H22O6

C 26H26O13

C 22H24O9

C 23H26O9

C 32H28O14

C 20H22O8

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KL

MN

O

Aisoorientin

210158401015840-O

-apiofuranoside1B

vitexin210158401015840-O

-xyloside2


D6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4E

apigenin


e)5

Fcerarvensin

6G

572101584031015840-te

trahydroxyflavone

7H5

2101584041015840-tr

ihydroxy-68-dim

etho

xyflavone

(rehderia

ninI)

8Imachaerol

B9

Jbrosim

acutin

B10K

isorham

netin

3-[610158401015840-(2-(E)-bu

teno

yl)-glucoside]

11L

3567310158404101584051015840-

heptam

etho

xyflavone

12M

5-hydroxy-78-dim

etho

xyflavano

ne5-rham

nosid


marylglucoside)

14O


5


O

O

HO

OHO

HO

HO

OH

OH

OH

OH

OH

OO

1

O

O

OOO

2

O

O

O

3

O

O

O

O

4

OH

O

OOH

OHO

OO

OO

O 5

OH

O

O

OHO

HO

OHHOHO

6

O

OOH

HO

7

OH

OH O

OOH

HO

8

OHO

O

HO

HO

9

OHO

HO

OH

OH

10

HO

OH

OHOH

OH

OH

OH

O

O

O

O11

HO

HO

HOHO

HO

HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

CH2OH

OH

OH

OH OHOH

OH

OH

OHCH2OH

CH2

OCH3

OCH3

OCH3

OCH3

OCH3

CH3

CH3

CH3

OH

H3CO

H3C

H3CO

CH2

CH3

O

Figure 7 Continued


O

O

12

O

O

HO

O

OHHO

HO

13

O

OH

HO

O

HO

HOOH

14

O

OH

HO

15

OH O

OO

HOOH

OH

OO

O

O

O

OO

O

O

O

HO

16

O OHO

OH

OH

OH

O O

CO

O

HO

OH

OH

O

OO

HO

HO

HOHO

HO

HO

O

OH

HO

17

HO

OH

OH

O

O

OOH

HOHO

HO

OHO

HO

HO

HO

O

O

18

O

OH

OH

OH

OHO

HO

HO

OH

19

O

HO

HO

OH

H3CO

H3CO

OCH3

OCH3

OCH3

OCH3

OCH3

H3C

OCH3

OCH3

H3C

CH2

CH2

CH3

CH3

CH3

CH3

CH2

2H

H3C

O

OO

O

Figure 7 Continued


OO

OH

OH OH

O

20

OHO

HO

HO

HO

O

OH

O

21

OHOOC

HO

HO

HO

OH

HO

O

O22

OH

O O

O

OO

O

23

HO

HO

HO

HO

HO

HO

OH

OH

OHOH

OH

HOOC

OH

H2C O

O




Sequence 1 2 3 4 5 6 7119905119877


26H28O15

C21H20O11

C25H26O13

C25H24O12

C32H34O18

C27H30O16

C26H28O14






Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O


8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol


6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















Wound contraction


Initial wound size

times 100

(1)





0

20

40

60

80

100

(Days)

w

ound

cont

ract

ion

0 5 10 15 20










Untreated

Standard

Cream

Aqueous fraction

n-Butanol fraction





50



Table 1 EC50








Table2Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringexcisio

nwou

ndmod

el

Days

Day

0(beforetreatment)

36

912

1517

Group

1(ethylacetate)

229plusmn14

7(000)


9025plusmn17

0lowast(6058)

6150plusmn18

4lowast(7314)


8)000plusmn000

(100)

000plusmn000

(100)

Group

2(n-butanol)

2312

5plusmn330

(000)

147plusmn12

2lowastlowast(3643)

1175plusmn10

4(491

8)48plusmn12

2lowastlowast(792


000plusmn000

(100)

000plusmn000

(100)

Group

3(aqu

eous)

2335plusmn589

(000)


118plusmn336

(494


7425plusmn19

3(6820)


9)000plusmn000

(100)

Group

4(cream

)22825plusmn423

(000)

196plusmn294

(1412


88plusmn12

2(614

4)595plusmn16

5lowastlowast(7393)


2)000plusmn000

(100)

Group

5(con

trol)

2292

5plusmn245

(000)

20325plusmn249

(114

3)118plusmn313

(4852)

9375plusmn375

(591

0)845plusmn35(6314)

54plusmn408

(7644

)305plusmn386

(8669)

Group

6(stand

ard)

2312

9plusmn473

(000)

14825plusmn19

3lowastlowast(3589)


0)44



000plusmn000

(100)

000plusmn000

(100)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)



002040608

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(a)

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

01234

times106



(b)


0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106


(a)

01234



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106

(b)


02468


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(a)

0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(b)













Table4Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringbu

rnwou

ndmod

elDays

Day

0(beforetreatment)

36

912

1517

19Group

1(ethylacetate)

499plusmn353

(000)

2917






000plusmn000

(000)

000plusmn000

(000)

Group

2(n-butanol)

47675plusmn92

5(000)

3717






6)000plusmn000

(000)

000plusmn000

(000)

Group

3(aqu

eous)

4897

5plusmn18

4(000)

3112


)1772


815plusmn15


485plusmn15

5lowastlowast(9009)

000plusmn000

(000)

000plusmn000

(000)

000plusmn000

(000)

Group

4(stand

ard)

492plusmn402

(000)


3175plusmn337

(3546




000plusmn000

(000)

000plusmn000

(000)

Group

5(con

trol)

4965plusmn560

(000)

42425plusmn840

(1455)

324plusmn74

2(3474)

3235plusmn75

(3484)

2072

5plusmn592

(5825)

1195plusmn613

(7593)

565plusmn575

(8862)

2675plusmn550

(9461)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)


Table5Listof

flavono

idstentativ

elyidentifi

edin

ethylacetatefractio

ns

Characteris

tics

Ethylacetatefractio

nMS+M

SminusSequ

ence

12

34

56

78

910

1112

1314

15119905119877

906

9333

9598

9694

10354

10602

11984

12816

12953

15021

11308

11316

1146

11857

14089

[MminusH](119898119911)

5801439

5641492

4481018

5341379

5161279

4020963

28604

853300746

3621373

3581422

5461368

4321426

4461597

6361493

3901325

Error(pp

m)

minus19

1minus23

minus27

minus10

5minus216

minus305

minus249

minus19

minus19

5minus15

910

3minus13

1minus459

minus226

minus273

Molecular

form

ula

C 26H28O15

C 26H28O14

C 21H20O11

C 25H26O13

C 25H24O12

C 20H18O9

C 15H10O6

C 17H14O7

C 19H22O7

C 20H22O6

C 26H26O13

C 22H24O9

C 23H26O9

C 32H28O14

C 20H22O8

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KL

MN

O

Aisoorientin

210158401015840-O

-apiofuranoside1B

vitexin210158401015840-O

-xyloside2


D6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4E

apigenin


e)5

Fcerarvensin

6G

572101584031015840-te

trahydroxyflavone

7H5

2101584041015840-tr

ihydroxy-68-dim

etho

xyflavone

(rehderia

ninI)

8Imachaerol

B9

Jbrosim

acutin

B10K

isorham

netin

3-[610158401015840-(2-(E)-bu

teno

yl)-glucoside]

11L

3567310158404101584051015840-

heptam

etho

xyflavone

12M

5-hydroxy-78-dim

etho

xyflavano

ne5-rham

nosid


marylglucoside)

14O


5


O

O

HO

OHO

HO

HO

OH

OH

OH

OH

OH

OO

1

O

O

OOO

2

O

O

O

3

O

O

O

O

4

OH

O

OOH

OHO

OO

OO

O 5

OH

O

O

OHO

HO

OHHOHO

6

O

OOH

HO

7

OH

OH O

OOH

HO

8

OHO

O

HO

HO

9

OHO

HO

OH

OH

10

HO

OH

OHOH

OH

OH

OH

O

O

O

O11

HO

HO

HOHO

HO

HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

CH2OH

OH

OH

OH OHOH

OH

OH

OHCH2OH

CH2

OCH3

OCH3

OCH3

OCH3

OCH3

CH3

CH3

CH3

OH

H3CO

H3C

H3CO

CH2

CH3

O

Figure 7 Continued


O

O

12

O

O

HO

O

OHHO

HO

13

O

OH

HO

O

HO

HOOH

14

O

OH

HO

15

OH O

OO

HOOH

OH

OO

O

O

O

OO

O

O

O

HO

16

O OHO

OH

OH

OH

O O

CO

O

HO

OH

OH

O

OO

HO

HO

HOHO

HO

HO

O

OH

HO

17

HO

OH

OH

O

O

OOH

HOHO

HO

OHO

HO

HO

HO

O

O

18

O

OH

OH

OH

OHO

HO

HO

OH

19

O

HO

HO

OH

H3CO

H3CO

OCH3

OCH3

OCH3

OCH3

OCH3

H3C

OCH3

OCH3

H3C

CH2

CH2

CH3

CH3

CH3

CH3

CH2

2H

H3C

O

OO

O

Figure 7 Continued


OO

OH

OH OH

O

20

OHO

HO

HO

HO

O

OH

O

21

OHOOC

HO

HO

HO

OH

HO

O

O22

OH

O O

O

OO

O

23

HO

HO

HO

HO

HO

HO

OH

OH

OHOH

OH

HOOC

OH

H2C O

O




Sequence 1 2 3 4 5 6 7119905119877


26H28O15

C21H20O11

C25H26O13

C25H24O12

C32H34O18

C27H30O16

C26H28O14






Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O


8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol


6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


















Untreated

Standard

Cream

Aqueous fraction

n-Butanol fraction





50



Table 1 EC50








Table2Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringexcisio

nwou

ndmod

el

Days

Day

0(beforetreatment)

36

912

1517

Group

1(ethylacetate)

229plusmn14

7(000)


9025plusmn17

0lowast(6058)

6150plusmn18

4lowast(7314)


8)000plusmn000

(100)

000plusmn000

(100)

Group

2(n-butanol)

2312

5plusmn330

(000)

147plusmn12

2lowastlowast(3643)

1175plusmn10

4(491

8)48plusmn12

2lowastlowast(792


000plusmn000

(100)

000plusmn000

(100)

Group

3(aqu

eous)

2335plusmn589

(000)


118plusmn336

(494


7425plusmn19

3(6820)


9)000plusmn000

(100)

Group

4(cream

)22825plusmn423

(000)

196plusmn294

(1412


88plusmn12

2(614

4)595plusmn16

5lowastlowast(7393)


2)000plusmn000

(100)

Group

5(con

trol)

2292

5plusmn245

(000)

20325plusmn249

(114

3)118plusmn313

(4852)

9375plusmn375

(591

0)845plusmn35(6314)

54plusmn408

(7644

)305plusmn386

(8669)

Group

6(stand

ard)

2312

9plusmn473

(000)

14825plusmn19

3lowastlowast(3589)


0)44



000plusmn000

(100)

000plusmn000

(100)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)



002040608

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(a)

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

01234

times106



(b)


0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106


(a)

01234



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106

(b)


02468


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(a)

0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(b)













Table4Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringbu

rnwou

ndmod

elDays

Day

0(beforetreatment)

36

912

1517

19Group

1(ethylacetate)

499plusmn353

(000)

2917






000plusmn000

(000)

000plusmn000

(000)

Group

2(n-butanol)

47675plusmn92

5(000)

3717






6)000plusmn000

(000)

000plusmn000

(000)

Group

3(aqu

eous)

4897

5plusmn18

4(000)

3112


)1772


815plusmn15


485plusmn15

5lowastlowast(9009)

000plusmn000

(000)

000plusmn000

(000)

000plusmn000

(000)

Group

4(stand

ard)

492plusmn402

(000)


3175plusmn337

(3546




000plusmn000

(000)

000plusmn000

(000)

Group

5(con

trol)

4965plusmn560

(000)

42425plusmn840

(1455)

324plusmn74

2(3474)

3235plusmn75

(3484)

2072

5plusmn592

(5825)

1195plusmn613

(7593)

565plusmn575

(8862)

2675plusmn550

(9461)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)


Table5Listof

flavono

idstentativ

elyidentifi

edin

ethylacetatefractio

ns

Characteris

tics

Ethylacetatefractio

nMS+M

SminusSequ

ence

12

34

56

78

910

1112

1314

15119905119877

906

9333

9598

9694

10354

10602

11984

12816

12953

15021

11308

11316

1146

11857

14089

[MminusH](119898119911)

5801439

5641492

4481018

5341379

5161279

4020963

28604

853300746

3621373

3581422

5461368

4321426

4461597

6361493

3901325

Error(pp

m)

minus19

1minus23

minus27

minus10

5minus216

minus305

minus249

minus19

minus19

5minus15

910

3minus13

1minus459

minus226

minus273

Molecular

form

ula

C 26H28O15

C 26H28O14

C 21H20O11

C 25H26O13

C 25H24O12

C 20H18O9

C 15H10O6

C 17H14O7

C 19H22O7

C 20H22O6

C 26H26O13

C 22H24O9

C 23H26O9

C 32H28O14

C 20H22O8

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KL

MN

O

Aisoorientin

210158401015840-O

-apiofuranoside1B

vitexin210158401015840-O

-xyloside2


D6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4E

apigenin


e)5

Fcerarvensin

6G

572101584031015840-te

trahydroxyflavone

7H5

2101584041015840-tr

ihydroxy-68-dim

etho

xyflavone

(rehderia

ninI)

8Imachaerol

B9

Jbrosim

acutin

B10K

isorham

netin

3-[610158401015840-(2-(E)-bu

teno

yl)-glucoside]

11L

3567310158404101584051015840-

heptam

etho

xyflavone

12M

5-hydroxy-78-dim

etho

xyflavano

ne5-rham

nosid


marylglucoside)

14O


5


O

O

HO

OHO

HO

HO

OH

OH

OH

OH

OH

OO

1

O

O

OOO

2

O

O

O

3

O

O

O

O

4

OH

O

OOH

OHO

OO

OO

O 5

OH

O

O

OHO

HO

OHHOHO

6

O

OOH

HO

7

OH

OH O

OOH

HO

8

OHO

O

HO

HO

9

OHO

HO

OH

OH

10

HO

OH

OHOH

OH

OH

OH

O

O

O

O11

HO

HO

HOHO

HO

HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

CH2OH

OH

OH

OH OHOH

OH

OH

OHCH2OH

CH2

OCH3

OCH3

OCH3

OCH3

OCH3

CH3

CH3

CH3

OH

H3CO

H3C

H3CO

CH2

CH3

O

Figure 7 Continued


O

O

12

O

O

HO

O

OHHO

HO

13

O

OH

HO

O

HO

HOOH

14

O

OH

HO

15

OH O

OO

HOOH

OH

OO

O

O

O

OO

O

O

O

HO

16

O OHO

OH

OH

OH

O O

CO

O

HO

OH

OH

O

OO

HO

HO

HOHO

HO

HO

O

OH

HO

17

HO

OH

OH

O

O

OOH

HOHO

HO

OHO

HO

HO

HO

O

O

18

O

OH

OH

OH

OHO

HO

HO

OH

19

O

HO

HO

OH

H3CO

H3CO

OCH3

OCH3

OCH3

OCH3

OCH3

H3C

OCH3

OCH3

H3C

CH2

CH2

CH3

CH3

CH3

CH3

CH2

2H

H3C

O

OO

O

Figure 7 Continued


OO

OH

OH OH

O

20

OHO

HO

HO

HO

O

OH

O

21

OHOOC

HO

HO

HO

OH

HO

O

O22

OH

O O

O

OO

O

23

HO

HO

HO

HO

HO

HO

OH

OH

OHOH

OH

HOOC

OH

H2C O

O




Sequence 1 2 3 4 5 6 7119905119877


26H28O15

C21H20O11

C25H26O13

C25H24O12

C32H34O18

C27H30O16

C26H28O14






Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O


8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol


6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table2Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringexcisio

nwou

ndmod

el

Days

Day

0(beforetreatment)

36

912

1517

Group

1(ethylacetate)

229plusmn14

7(000)


9025plusmn17

0lowast(6058)

6150plusmn18

4lowast(7314)


8)000plusmn000

(100)

000plusmn000

(100)

Group

2(n-butanol)

2312

5plusmn330

(000)

147plusmn12

2lowastlowast(3643)

1175plusmn10

4(491

8)48plusmn12

2lowastlowast(792


000plusmn000

(100)

000plusmn000

(100)

Group

3(aqu

eous)

2335plusmn589

(000)


118plusmn336

(494


7425plusmn19

3(6820)


9)000plusmn000

(100)

Group

4(cream

)22825plusmn423

(000)

196plusmn294

(1412


88plusmn12

2(614

4)595plusmn16

5lowastlowast(7393)


2)000plusmn000

(100)

Group

5(con

trol)

2292

5plusmn245

(000)

20325plusmn249

(114

3)118plusmn313

(4852)

9375plusmn375

(591

0)845plusmn35(6314)

54plusmn408

(7644

)305plusmn386

(8669)

Group

6(stand

ard)

2312

9plusmn473

(000)

14825plusmn19

3lowastlowast(3589)


0)44



000plusmn000

(100)

000plusmn000

(100)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)



002040608

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(a)

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

01234

times106



(b)


0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106


(a)

01234



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106

(b)


02468


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(a)

0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(b)













Table4Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringbu

rnwou

ndmod

elDays

Day

0(beforetreatment)

36

912

1517

19Group

1(ethylacetate)

499plusmn353

(000)

2917






000plusmn000

(000)

000plusmn000

(000)

Group

2(n-butanol)

47675plusmn92

5(000)

3717






6)000plusmn000

(000)

000plusmn000

(000)

Group

3(aqu

eous)

4897

5plusmn18

4(000)

3112


)1772


815plusmn15


485plusmn15

5lowastlowast(9009)

000plusmn000

(000)

000plusmn000

(000)

000plusmn000

(000)

Group

4(stand

ard)

492plusmn402

(000)


3175plusmn337

(3546




000plusmn000

(000)

000plusmn000

(000)

Group

5(con

trol)

4965plusmn560

(000)

42425plusmn840

(1455)

324plusmn74

2(3474)

3235plusmn75

(3484)

2072

5plusmn592

(5825)

1195plusmn613

(7593)

565plusmn575

(8862)

2675plusmn550

(9461)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)


Table5Listof

flavono

idstentativ

elyidentifi

edin

ethylacetatefractio

ns

Characteris

tics

Ethylacetatefractio

nMS+M

SminusSequ

ence

12

34

56

78

910

1112

1314

15119905119877

906

9333

9598

9694

10354

10602

11984

12816

12953

15021

11308

11316

1146

11857

14089

[MminusH](119898119911)

5801439

5641492

4481018

5341379

5161279

4020963

28604

853300746

3621373

3581422

5461368

4321426

4461597

6361493

3901325

Error(pp

m)

minus19

1minus23

minus27

minus10

5minus216

minus305

minus249

minus19

minus19

5minus15

910

3minus13

1minus459

minus226

minus273

Molecular

form

ula

C 26H28O15

C 26H28O14

C 21H20O11

C 25H26O13

C 25H24O12

C 20H18O9

C 15H10O6

C 17H14O7

C 19H22O7

C 20H22O6

C 26H26O13

C 22H24O9

C 23H26O9

C 32H28O14

C 20H22O8

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KL

MN

O

Aisoorientin

210158401015840-O

-apiofuranoside1B

vitexin210158401015840-O

-xyloside2


D6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4E

apigenin


e)5

Fcerarvensin

6G

572101584031015840-te

trahydroxyflavone

7H5

2101584041015840-tr

ihydroxy-68-dim

etho

xyflavone

(rehderia

ninI)

8Imachaerol

B9

Jbrosim

acutin

B10K

isorham

netin

3-[610158401015840-(2-(E)-bu

teno

yl)-glucoside]

11L

3567310158404101584051015840-

heptam

etho

xyflavone

12M

5-hydroxy-78-dim

etho

xyflavano

ne5-rham

nosid


marylglucoside)

14O


5


O

O

HO

OHO

HO

HO

OH

OH

OH

OH

OH

OO

1

O

O

OOO

2

O

O

O

3

O

O

O

O

4

OH

O

OOH

OHO

OO

OO

O 5

OH

O

O

OHO

HO

OHHOHO

6

O

OOH

HO

7

OH

OH O

OOH

HO

8

OHO

O

HO

HO

9

OHO

HO

OH

OH

10

HO

OH

OHOH

OH

OH

OH

O

O

O

O11

HO

HO

HOHO

HO

HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

CH2OH

OH

OH

OH OHOH

OH

OH

OHCH2OH

CH2

OCH3

OCH3

OCH3

OCH3

OCH3

CH3

CH3

CH3

OH

H3CO

H3C

H3CO

CH2

CH3

O

Figure 7 Continued


O

O

12

O

O

HO

O

OHHO

HO

13

O

OH

HO

O

HO

HOOH

14

O

OH

HO

15

OH O

OO

HOOH

OH

OO

O

O

O

OO

O

O

O

HO

16

O OHO

OH

OH

OH

O O

CO

O

HO

OH

OH

O

OO

HO

HO

HOHO

HO

HO

O

OH

HO

17

HO

OH

OH

O

O

OOH

HOHO

HO

OHO

HO

HO

HO

O

O

18

O

OH

OH

OH

OHO

HO

HO

OH

19

O

HO

HO

OH

H3CO

H3CO

OCH3

OCH3

OCH3

OCH3

OCH3

H3C

OCH3

OCH3

H3C

CH2

CH2

CH3

CH3

CH3

CH3

CH2

2H

H3C

O

OO

O

Figure 7 Continued


OO

OH

OH OH

O

20

OHO

HO

HO

HO

O

OH

O

21

OHOOC

HO

HO

HO

OH

HO

O

O22

OH

O O

O

OO

O

23

HO

HO

HO

HO

HO

HO

OH

OH

OHOH

OH

HOOC

OH

H2C O

O




Sequence 1 2 3 4 5 6 7119905119877


26H28O15

C21H20O11

C25H26O13

C25H24O12

C32H34O18

C27H30O16

C26H28O14






Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O


8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol


6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


















002040608

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(a)

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

01234

times106



(b)


0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106


(a)

01234



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

times106

(b)


02468


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(a)

0123456


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24


(b)













Table4Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringbu

rnwou

ndmod

elDays

Day

0(beforetreatment)

36

912

1517

19Group

1(ethylacetate)

499plusmn353

(000)

2917






000plusmn000

(000)

000plusmn000

(000)

Group

2(n-butanol)

47675plusmn92

5(000)

3717






6)000plusmn000

(000)

000plusmn000

(000)

Group

3(aqu

eous)

4897

5plusmn18

4(000)

3112


)1772


815plusmn15


485plusmn15

5lowastlowast(9009)

000plusmn000

(000)

000plusmn000

(000)

000plusmn000

(000)

Group

4(stand

ard)

492plusmn402

(000)


3175plusmn337

(3546




000plusmn000

(000)

000plusmn000

(000)

Group

5(con

trol)

4965plusmn560

(000)

42425plusmn840

(1455)

324plusmn74

2(3474)

3235plusmn75

(3484)

2072

5plusmn592

(5825)

1195plusmn613

(7593)

565plusmn575

(8862)

2675plusmn550

(9461)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)


Table5Listof

flavono

idstentativ

elyidentifi

edin

ethylacetatefractio

ns

Characteris

tics

Ethylacetatefractio

nMS+M

SminusSequ

ence

12

34

56

78

910

1112

1314

15119905119877

906

9333

9598

9694

10354

10602

11984

12816

12953

15021

11308

11316

1146

11857

14089

[MminusH](119898119911)

5801439

5641492

4481018

5341379

5161279

4020963

28604

853300746

3621373

3581422

5461368

4321426

4461597

6361493

3901325

Error(pp

m)

minus19

1minus23

minus27

minus10

5minus216

minus305

minus249

minus19

minus19

5minus15

910

3minus13

1minus459

minus226

minus273

Molecular

form

ula

C 26H28O15

C 26H28O14

C 21H20O11

C 25H26O13

C 25H24O12

C 20H18O9

C 15H10O6

C 17H14O7

C 19H22O7

C 20H22O6

C 26H26O13

C 22H24O9

C 23H26O9

C 32H28O14

C 20H22O8

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KL

MN

O

Aisoorientin

210158401015840-O

-apiofuranoside1B

vitexin210158401015840-O

-xyloside2


D6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4E

apigenin


e)5

Fcerarvensin

6G

572101584031015840-te

trahydroxyflavone

7H5

2101584041015840-tr

ihydroxy-68-dim

etho

xyflavone

(rehderia

ninI)

8Imachaerol

B9

Jbrosim

acutin

B10K

isorham

netin

3-[610158401015840-(2-(E)-bu

teno

yl)-glucoside]

11L

3567310158404101584051015840-

heptam

etho

xyflavone

12M

5-hydroxy-78-dim

etho

xyflavano

ne5-rham

nosid


marylglucoside)

14O


5


O

O

HO

OHO

HO

HO

OH

OH

OH

OH

OH

OO

1

O

O

OOO

2

O

O

O

3

O

O

O

O

4

OH

O

OOH

OHO

OO

OO

O 5

OH

O

O

OHO

HO

OHHOHO

6

O

OOH

HO

7

OH

OH O

OOH

HO

8

OHO

O

HO

HO

9

OHO

HO

OH

OH

10

HO

OH

OHOH

OH

OH

OH

O

O

O

O11

HO

HO

HOHO

HO

HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

CH2OH

OH

OH

OH OHOH

OH

OH

OHCH2OH

CH2

OCH3

OCH3

OCH3

OCH3

OCH3

CH3

CH3

CH3

OH

H3CO

H3C

H3CO

CH2

CH3

O

Figure 7 Continued


O

O

12

O

O

HO

O

OHHO

HO

13

O

OH

HO

O

HO

HOOH

14

O

OH

HO

15

OH O

OO

HOOH

OH

OO

O

O

O

OO

O

O

O

HO

16

O OHO

OH

OH

OH

O O

CO

O

HO

OH

OH

O

OO

HO

HO

HOHO

HO

HO

O

OH

HO

17

HO

OH

OH

O

O

OOH

HOHO

HO

OHO

HO

HO

HO

O

O

18

O

OH

OH

OH

OHO

HO

HO

OH

19

O

HO

HO

OH

H3CO

H3CO

OCH3

OCH3

OCH3

OCH3

OCH3

H3C

OCH3

OCH3

H3C

CH2

CH2

CH3

CH3

CH3

CH3

CH2

2H

H3C

O

OO

O

Figure 7 Continued


OO

OH

OH OH

O

20

OHO

HO

HO

HO

O

OH

O

21

OHOOC

HO

HO

HO

OH

HO

O

O22

OH

O O

O

OO

O

23

HO

HO

HO

HO

HO

HO

OH

OH

OHOH

OH

HOOC

OH

H2C O

O




Sequence 1 2 3 4 5 6 7119905119877


26H28O15

C21H20O11

C25H26O13

C25H24O12

C32H34O18

C27H30O16

C26H28O14






Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O


8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol


6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table4Meanarea

ofwou

nd(m

m2)a

ndwou

ndcontractiondu

ringbu

rnwou

ndmod

elDays

Day

0(beforetreatment)

36

912

1517

19Group

1(ethylacetate)

499plusmn353

(000)

2917






000plusmn000

(000)

000plusmn000

(000)

Group

2(n-butanol)

47675plusmn92

5(000)

3717






6)000plusmn000

(000)

000plusmn000

(000)

Group

3(aqu

eous)

4897

5plusmn18

4(000)

3112


)1772


815plusmn15


485plusmn15

5lowastlowast(9009)

000plusmn000

(000)

000plusmn000

(000)

000plusmn000

(000)

Group

4(stand

ard)

492plusmn402

(000)


3175plusmn337

(3546




000plusmn000

(000)

000plusmn000

(000)

Group

5(con

trol)

4965plusmn560

(000)

42425plusmn840

(1455)

324plusmn74

2(3474)

3235plusmn75

(3484)

2072

5plusmn592

(5825)

1195plusmn613

(7593)

565plusmn575

(8862)

2675plusmn550

(9461)

Values

aremeanplusmnSE

M(percent)o

f4ratsin

each

grou


respectiv

edaycontrolg

roup

(statistic

alanalysiswas

done

byon

e-way

analysisof

varia

nce

follo

wed

byDun

nettrsquostestfor

multip

lecomparis

ons)


Table5Listof

flavono

idstentativ

elyidentifi

edin

ethylacetatefractio

ns

Characteris

tics

Ethylacetatefractio

nMS+M

SminusSequ

ence

12

34

56

78

910

1112

1314

15119905119877

906

9333

9598

9694

10354

10602

11984

12816

12953

15021

11308

11316

1146

11857

14089

[MminusH](119898119911)

5801439

5641492

4481018

5341379

5161279

4020963

28604

853300746

3621373

3581422

5461368

4321426

4461597

6361493

3901325

Error(pp

m)

minus19

1minus23

minus27

minus10

5minus216

minus305

minus249

minus19

minus19

5minus15

910

3minus13

1minus459

minus226

minus273

Molecular

form

ula

C 26H28O15

C 26H28O14

C 21H20O11

C 25H26O13

C 25H24O12

C 20H18O9

C 15H10O6

C 17H14O7

C 19H22O7

C 20H22O6

C 26H26O13

C 22H24O9

C 23H26O9

C 32H28O14

C 20H22O8

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KL

MN

O

Aisoorientin

210158401015840-O

-apiofuranoside1B

vitexin210158401015840-O

-xyloside2


D6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4E

apigenin


e)5

Fcerarvensin

6G

572101584031015840-te

trahydroxyflavone

7H5

2101584041015840-tr

ihydroxy-68-dim

etho

xyflavone

(rehderia

ninI)

8Imachaerol

B9

Jbrosim

acutin

B10K

isorham

netin

3-[610158401015840-(2-(E)-bu

teno

yl)-glucoside]

11L

3567310158404101584051015840-

heptam

etho

xyflavone

12M

5-hydroxy-78-dim

etho

xyflavano

ne5-rham

nosid


marylglucoside)

14O


5


O

O

HO

OHO

HO

HO

OH

OH

OH

OH

OH

OO

1

O

O

OOO

2

O

O

O

3

O

O

O

O

4

OH

O

OOH

OHO

OO

OO

O 5

OH

O

O

OHO

HO

OHHOHO

6

O

OOH

HO

7

OH

OH O

OOH

HO

8

OHO

O

HO

HO

9

OHO

HO

OH

OH

10

HO

OH

OHOH

OH

OH

OH

O

O

O

O11

HO

HO

HOHO

HO

HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

CH2OH

OH

OH

OH OHOH

OH

OH

OHCH2OH

CH2

OCH3

OCH3

OCH3

OCH3

OCH3

CH3

CH3

CH3

OH

H3CO

H3C

H3CO

CH2

CH3

O

Figure 7 Continued


O

O

12

O

O

HO

O

OHHO

HO

13

O

OH

HO

O

HO

HOOH

14

O

OH

HO

15

OH O

OO

HOOH

OH

OO

O

O

O

OO

O

O

O

HO

16

O OHO

OH

OH

OH

O O

CO

O

HO

OH

OH

O

OO

HO

HO

HOHO

HO

HO

O

OH

HO

17

HO

OH

OH

O

O

OOH

HOHO

HO

OHO

HO

HO

HO

O

O

18

O

OH

OH

OH

OHO

HO

HO

OH

19

O

HO

HO

OH

H3CO

H3CO

OCH3

OCH3

OCH3

OCH3

OCH3

H3C

OCH3

OCH3

H3C

CH2

CH2

CH3

CH3

CH3

CH3

CH2

2H

H3C

O

OO

O

Figure 7 Continued


OO

OH

OH OH

O

20

OHO

HO

HO

HO

O

OH

O

21

OHOOC

HO

HO

HO

OH

HO

O

O22

OH

O O

O

OO

O

23

HO

HO

HO

HO

HO

HO

OH

OH

OHOH

OH

HOOC

OH

H2C O

O




Sequence 1 2 3 4 5 6 7119905119877


26H28O15

C21H20O11

C25H26O13

C25H24O12

C32H34O18

C27H30O16

C26H28O14






Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O


8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol


6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table5Listof

flavono

idstentativ

elyidentifi

edin

ethylacetatefractio

ns

Characteris

tics

Ethylacetatefractio

nMS+M

SminusSequ

ence

12

34

56

78

910

1112

1314

15119905119877

906

9333

9598

9694

10354

10602

11984

12816

12953

15021

11308

11316

1146

11857

14089

[MminusH](119898119911)

5801439

5641492

4481018

5341379

5161279

4020963

28604

853300746

3621373

3581422

5461368

4321426

4461597

6361493

3901325

Error(pp

m)

minus19

1minus23

minus27

minus10

5minus216

minus305

minus249

minus19

minus19

5minus15

910

3minus13

1minus459

minus226

minus273

Molecular

form

ula

C 26H28O15

C 26H28O14

C 21H20O11

C 25H26O13

C 25H24O12

C 20H18O9

C 15H10O6

C 17H14O7

C 19H22O7

C 20H22O6

C 26H26O13

C 22H24O9

C 23H26O9

C 32H28O14

C 20H22O8

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KL

MN

O

Aisoorientin

210158401015840-O

-apiofuranoside1B

vitexin210158401015840-O

-xyloside2


D6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4E

apigenin


e)5

Fcerarvensin

6G

572101584031015840-te

trahydroxyflavone

7H5

2101584041015840-tr

ihydroxy-68-dim

etho

xyflavone

(rehderia

ninI)

8Imachaerol

B9

Jbrosim

acutin

B10K

isorham

netin

3-[610158401015840-(2-(E)-bu

teno

yl)-glucoside]

11L

3567310158404101584051015840-

heptam

etho

xyflavone

12M

5-hydroxy-78-dim

etho

xyflavano

ne5-rham

nosid


marylglucoside)

14O


5


O

O

HO

OHO

HO

HO

OH

OH

OH

OH

OH

OO

1

O

O

OOO

2

O

O

O

3

O

O

O

O

4

OH

O

OOH

OHO

OO

OO

O 5

OH

O

O

OHO

HO

OHHOHO

6

O

OOH

HO

7

OH

OH O

OOH

HO

8

OHO

O

HO

HO

9

OHO

HO

OH

OH

10

HO

OH

OHOH

OH

OH

OH

O

O

O

O11

HO

HO

HOHO

HO

HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

CH2OH

OH

OH

OH OHOH

OH

OH

OHCH2OH

CH2

OCH3

OCH3

OCH3

OCH3

OCH3

CH3

CH3

CH3

OH

H3CO

H3C

H3CO

CH2

CH3

O

Figure 7 Continued


O

O

12

O

O

HO

O

OHHO

HO

13

O

OH

HO

O

HO

HOOH

14

O

OH

HO

15

OH O

OO

HOOH

OH

OO

O

O

O

OO

O

O

O

HO

16

O OHO

OH

OH

OH

O O

CO

O

HO

OH

OH

O

OO

HO

HO

HOHO

HO

HO

O

OH

HO

17

HO

OH

OH

O

O

OOH

HOHO

HO

OHO

HO

HO

HO

O

O

18

O

OH

OH

OH

OHO

HO

HO

OH

19

O

HO

HO

OH

H3CO

H3CO

OCH3

OCH3

OCH3

OCH3

OCH3

H3C

OCH3

OCH3

H3C

CH2

CH2

CH3

CH3

CH3

CH3

CH2

2H

H3C

O

OO

O

Figure 7 Continued


OO

OH

OH OH

O

20

OHO

HO

HO

HO

O

OH

O

21

OHOOC

HO

HO

HO

OH

HO

O

O22

OH

O O

O

OO

O

23

HO

HO

HO

HO

HO

HO

OH

OH

OHOH

OH

HOOC

OH

H2C O

O




Sequence 1 2 3 4 5 6 7119905119877


26H28O15

C21H20O11

C25H26O13

C25H24O12

C32H34O18

C27H30O16

C26H28O14






Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O


8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol


6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















O

O

HO

OHO

HO

HO

OH

OH

OH

OH

OH

OO

1

O

O

OOO

2

O

O

O

3

O

O

O

O

4

OH

O

OOH

OHO

OO

OO

O 5

OH

O

O

OHO

HO

OHHOHO

6

O

OOH

HO

7

OH

OH O

OOH

HO

8

OHO

O

HO

HO

9

OHO

HO

OH

OH

10

HO

OH

OHOH

OH

OH

OH

O

O

O

O11

HO

HO

HOHO

HO

HO

HO

HO

HO

HO

HO

HO

OH

OH

OH

OH

OH

CH2OH

OH

OH

OH OHOH

OH

OH

OHCH2OH

CH2

OCH3

OCH3

OCH3

OCH3

OCH3

CH3

CH3

CH3

OH

H3CO

H3C

H3CO

CH2

CH3

O

Figure 7 Continued


O

O

12

O

O

HO

O

OHHO

HO

13

O

OH

HO

O

HO

HOOH

14

O

OH

HO

15

OH O

OO

HOOH

OH

OO

O

O

O

OO

O

O

O

HO

16

O OHO

OH

OH

OH

O O

CO

O

HO

OH

OH

O

OO

HO

HO

HOHO

HO

HO

O

OH

HO

17

HO

OH

OH

O

O

OOH

HOHO

HO

OHO

HO

HO

HO

O

O

18

O

OH

OH

OH

OHO

HO

HO

OH

19

O

HO

HO

OH

H3CO

H3CO

OCH3

OCH3

OCH3

OCH3

OCH3

H3C

OCH3

OCH3

H3C

CH2

CH2

CH3

CH3

CH3

CH3

CH2

2H

H3C

O

OO

O

Figure 7 Continued


OO

OH

OH OH

O

20

OHO

HO

HO

HO

O

OH

O

21

OHOOC

HO

HO

HO

OH

HO

O

O22

OH

O O

O

OO

O

23

HO

HO

HO

HO

HO

HO

OH

OH

OHOH

OH

HOOC

OH

H2C O

O




Sequence 1 2 3 4 5 6 7119905119877


26H28O15

C21H20O11

C25H26O13

C25H24O12

C32H34O18

C27H30O16

C26H28O14






Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O


8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol


6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















O

O

12

O

O

HO

O

OHHO

HO

13

O

OH

HO

O

HO

HOOH

14

O

OH

HO

15

OH O

OO

HOOH

OH

OO

O

O

O

OO

O

O

O

HO

16

O OHO

OH

OH

OH

O O

CO

O

HO

OH

OH

O

OO

HO

HO

HOHO

HO

HO

O

OH

HO

17

HO

OH

OH

O

O

OOH

HOHO

HO

OHO

HO

HO

HO

O

O

18

O

OH

OH

OH

OHO

HO

HO

OH

19

O

HO

HO

OH

H3CO

H3CO

OCH3

OCH3

OCH3

OCH3

OCH3

H3C

OCH3

OCH3

H3C

CH2

CH2

CH3

CH3

CH3

CH3

CH2

2H

H3C

O

OO

O

Figure 7 Continued


OO

OH

OH OH

O

20

OHO

HO

HO

HO

O

OH

O

21

OHOOC

HO

HO

HO

OH

HO

O

O22

OH

O O

O

OO

O

23

HO

HO

HO

HO

HO

HO

OH

OH

OHOH

OH

HOOC

OH

H2C O

O




Sequence 1 2 3 4 5 6 7119905119877


26H28O15

C21H20O11

C25H26O13

C25H24O12

C32H34O18

C27H30O16

C26H28O14






Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O


8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol


6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















OO

OH

OH OH

O

20

OHO

HO

HO

HO

O

OH

O

21

OHOOC

HO

HO

HO

OH

HO

O

O22

OH

O O

O

OO

O

23

HO

HO

HO

HO

HO

HO

OH

OH

OHOH

OH

HOOC

OH

H2C O

O




Sequence 1 2 3 4 5 6 7119905119877


26H28O15

C21H20O11

C25H26O13

C25H24O12

C32H34O18

C27H30O16

C26H28O14






Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O


8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol


6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table7Listof

flavono

idstentativ

elyidentifi

edin

aqueou

sfractions

Characteris

tics

Aqueou

sfractionMS+M

SminusSequ

ence

12

34

56

78

910

11119905 119877

8757

8944

908

9159

9325

9598

9714

10174

9444

1016

310365

[MminusH](119898119911)

7301724

610154

5801445

5641488

5941595

4481011

5341385

4620813

5501329

4620803

5161266

Error(pp

m)

296

minus10

5minus281

minus15

minus174

minus12

9minus208

minus328

minus115

minus10

9037

Molecular

form

ula

C 34H34O18

C 27H30O16

C 26H28O15

C 26H28O14

C 27H30O15

C 21H20O11

C 25H26O13

C 21H18O12

C 25H26O14

C 21H18O12

C 25H24O12

Prop

osed

compo

und

AB

CD

EF

GH

IJ

KAisorie

ntin

41015840-O

-glucosid

e210158401015840-O


8B

6-hydroxyluteolin

7-rutin

oside17C

isoorientin

210158401015840-O

-apiofuranoside1D

vitexin210158401015840-O

-xyloside2

Eisoorientin

7-O-rhamno

side19Fscutellarein

7-glucoside20

G6

8-D

i-C-beta-D-arabino

pyrano

sylap

igenin

4Hs

cutellarein

5-glucuron

ide21Ik

aempferol


6721015840-te

trahydroxyflavone

7-glucuron

ide23K

apigenin7-(210158401015840310158401015840-

diacetylglucoside)

5


Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Acknowledgments


References

































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















research article antioxidant and wound healing activity of

Documents