International Research Journal of Biological Sciences ___________________________________ ISSN 2278-3202 Vol. 2(1), 70-72, January (2013)Int. Res. J. Biological Sci. International Science Congress Association 70 Short CommunicationCharacterization of Black Chia Seed (Salvia hispanica L) and Oil and Quantification of -sitosterol Alonso-Caldern A.1*, Chvez-Bravo E.2, Rivera Antonio2, Montalvo-Paquini C.3, Arroyo-Tapia R.3, Monterrosas-Santamaria M.1, Jimnez-Salgado T.2 and Tapia-Hernndez A.2 1Facultad de Ingeniera Qumica de la Benemrita Universidad Autnoma de Puebla, MXICO 2Centro de Investigaciones en Ciencias Microbiolgicas del Instituto de Ciencias de la Benemrita Universidad Autnoma de Puebla, MXICO 3Universidad Politcnica de Puebla, MXICO Available online at: www.isca.in Received 4th December 2012, revised 21st December 2012, accepted 30th December 2012 AbstractTheimportanceofcorn,bean,amaranthandblackchiaseedintheAztecsdietiswelldocumentedinthehistorical FlorentineCodex.Blackchiaseedisagoodsourceofomega-3fattyacid,dietaryfiber,proteins,aswellasdiverse bioelementsandantioxidants.Theproximalchemicalanalysisobtainedfromblackchiaseedshows5.15%ofmoisture, 34.70% of fats, 4.10% of ashes, 17.90% of protein, 40.25% carbohydrates and 17.30% of raw fiber. The extraction technique permitted to obtain a 29% of oil with an iodine value of 197.68 cg/g, peroxides of2.67 meq/Kg, a refraction index of 1.46 and a density of 0.89 g/mL. The oils FTIR spectrum showed a peak at 3010.03 cm-1characteristic of a =CH stretching mode.Also,theprofileoffattyacidsperformedby,CG-MS,providesevidenceofthepresenceofthe(Z.Z.Z)9,12,15-octadecatrienoicacid(-linolenicacid,omega-3),-tocopherolaswellasphytosterols:campesterol,stigmasteroland-sitosterol. The latter was quantified obtaining a concentration of 1224.3 mg/L. Keywords: Campesterol, chia, stigmasterol, phytosterol, polyunsaturated, sitosterol. IntroductionBy the time of the Spanish conquest, Mesoamerican peoples had domesticatedatleast20botanicalspecieswithdifferentuses.Amongthose,amaranth,beans,blackchiaseedandcornstood outfromanutritionalstandpoint.Therelevanceofthosecrops intheAztecsdietisdocumentedinthehistoricalFlorentine Codex.Inparticular,blackchiaseedwasasimportantascorn and in some regions even more1. In the century 16th, the Codex Mendozaindicatesthat21ofthe38Aztecprovincialstates offeredchiaseedasannualtribute2.Moreover,ithasbeen determined that chia seed contains different oil amounts varying from32%to39% 3.Chiaseedoilalongwithlinseedoil,offer the highest natural percentage of omega-3 fatty acids, which are essentialinthehumandiet4.Omega-3fattyacidshelptomake upthephospholipidsthatarefundamentalcomponentsofcell membranes5. Recentworkhasshownthathensfedwithchiaseedproduced eggswithhigherlevelsofpolyunsaturatedlinolenicacidand lowercholesterol,palmitoleicandsaturatedacidlevelsintheir yolkswhencomparedwitheggsproducedfromhensundera controldiet 6.Chiaseedcanbeconsideredasagoodsourceof dietaryfiber,proteinandantioxidants.Chiaseedinaqueous medium is wrapped one at a mucilaginous polysaccharide which isexcellentfordigestion.Theconsumptionofdietaryfiber improvesfecalbolusformationandproperevacuationofstool, whichhelpspreventobesity,coloncancer,andhighlevelsof cholesterol and blood glucose7. Phytosterolsaresterolswithastructuresimilartothatof cholesterolwithsomemodifications.Thesemodifications involve the side chain and include the addition of a double bond and/or a methyl or ethyl group8. In general, vegetable oils (from corn,soybeanandchiaseed)containlargeamountsof polyunsaturatedfattyacidsandhavemorephytosterols9.From themorethan25differenttypesofphytosterolsthemost commonare:sitosterol,campesterolandstigmasterol, correspondingto65,30and3%ofthedailyintakeofeach respectively10.Someofthebenefitsthatareattributedtoadiet richinphytosterolsincludeanticancereffects8,anti-inflammatoryproperties11,antiulcereffects12,antioxidants effects13,bactericidalandantifungaleffects14.Theirabilityto controlcholesterolplasmalevelsinhypercholesterolimic patients was first described in 198311. In recent years, great deal ofinteresthasbeengiventotheroleof-sitosterolinthe protectionfromcoloncancer8andtheirabilitytoprevent complications from benign prostatic hyperplasia15,16. Therefore,theobjectiveofthepresentstudywastoidentify beneficialsubstancestohumanhealthandquantifythe-sitosterol from black chia seed, like related substance to prevent somediseaseslikecoloncancer,benignprostatichyperplasia and cholesterolemy.International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(1), 70-72, January (2013)Int. Res. J. Biological Sci.

International Science Congress Association 71 Material and Methods Proximal chemical analyses were performed on Black chia seed fromSanMateoCoatepecAtzitzihuacanPuebla,Mexico accordingtoAssociationofOfficialAnalyticalChemist (AOAC)techniques17.Theanalysisincludedmoisture(14,003 AOAC),ash(14,006AOAC),etherextract(7062AOAC), proteindetermination(2057AOAC),andtotalfiber(7068 AOAC).Fortheextraction,75 gramsof pulverized black chia seed wereplacedforfourhoursin a soxhletwith750mLofn-hexane (Merck). After the extraction, the n-hexane was removed by rotary evaporation at 60 C during 40 minutes. The iodine value (Cd 8-53 method)18 and peroxide index (Cd 1-25method)19oftheoilweredeterminedaccordingto the methodologies oftheAmericanOilChemistsSociety(AOCS, 2006).Theindexofrefractionwasmeasuredbyusingare fractometer(Reichentequipment,Arias500model)at25C. The density was obtained by the pycnometer method at 25C. InfraredspectrumwasobtainedinaPerkinElmerSpectrum OneFTIR Spectrometer in arange from 4000 to 400 cm-1. Gas chromatography(GC-MS)wasobtainedbyagas chromatograph(modelHP6890)withaselectivedetectorof masses,usingacapillarycolumn(modelHP5-MS)of30m long.Theinjectiontemperaturewas200Cinsplitmodewith 40:1 ratio. Thedetectortemperaturewas250C.Anincreaseof 194C was used during one minute until 250C at a rate of 30C perminute.Heliumgaswasusedinthemobilephaseanda selectivedetectorofmasses(model5973)withmassspectra library NIST 2.0 with manual injection. Finally, -sitosterol was quantifiedusingasitosterol-Bstandard(AmericanHerbal Pharmacopoeia).Alldeterminationswereperformedin triplicate. Results and Discussion Proximalchemicalanalysisfromblackchiaseedwas5.15%of moisture,34.70%offat,4.10%ofash,17.90%ofprotein, 40.25%ofcarbohydrateand17.30%ofrawfiber.The compounds inmajor proportionwere lipids, in accordance with previousstudiescorrespondtomainlyomega-3fattyacids. Also,itwascheckedaproteinintakeandtotalcarbohydrates, which almost 50% corresponding to raw fiber. Extractedblackchiaseedoilyieldwas29%.The physicochemicalcharacterizationindicatesaniodinevalueof 197.68cg/gcorrespondingtopolyunsaturatedfattyacids present,similartotheonereportedforcreolechiaof193 cgI2/g20.Blackchiaseedoilcontainshigheramountsof polyunsaturatedfattyacidsthanwithotheroilssuchascanola (110-126cgI2/g),safflower(136-148cgI2/g),olive(118139cgI2/g)andsoybean(118139cgI2/g)oils.Theperoxide valueobtainedwas2.64meq/Kg.Thevalueobtainedis relatively low to be unrefined oil, compared with themaximum value for refined soybean oilof 2meq/Kg (NMX-F-252-SFC1-2005). This low valueshowsthatnatural antioxidants thathave beenidentifiedsuchas-tocopherol,protectstheoxidationof omega-3fattyacids.Therefractionindexwas1.46, corresponding to polyunsaturated acids. Finally, the density was determined to be 0.89 g/mL. ThefunctionalgroupsidentifybyFTIRspectrum, correspondingtomoleculesfromunsaturatedfattyacids21,22 (table 1). Table-1 Functional groups identify by FTIR Functional groupsWavenumber (cm-1) = CH3010.03 C=O1744.27 The fatty acid profile obtained byCG-MS (table 2) showed the presenceofimportantcompoundsforhumanmetabolismthat are present in the black chia seed oil from San Mateo Coatepec Atzitzihuacan, Puebla. Among those compounds Z,Z,Z-9,12,15-octadecatrienoic acid (omega-3) was found with a retention time of 21.44 minutes. The presence of this compound has relevance becauseofthenumerousbenefitsassociatedwithits consumption. For example, medical and epidemiological studies haveshownthatconsuminglipidsrichinomega-3fattyacids reducetheriskofsufferingcoronaryheartdiseases.Another compoundfoundwas-tocopherolawitharetentiontimeof 25.71minutes.Anaturalantioxidant,-tocopherolplaysan importantroleintheprotectionofthepolyunsaturated compoundsfromoils.Phytosterolssuchascampesterol, stigmasterol and -sitosterol were found at 29.06 minutes, 29.73 minutesand31.05minutes,respectively.Theconcentrationof -sitosterolwasquantifiedas1224.3mg/L.Thepresenceof-sitosterol has been associated with prevention from prostatic and coloncancerandmayhelptoalleviatethesymptomsand urinaryflowproblemscausedbyanincreaseofthebenign prostatichyperplasia(BPH)anddecreasecholesterollevels (HDL). Table-2 Compounds identify by CG-MS CompoundsRetention time (minutes) Z,Z,Z-9,12,15-octadecatrienoic acid 21.44 -tocopherol25.71 campesterol29.06 stigmasterol29.73 -sitosterol31.05 Conclusion Proximalchemicalanalysisfromblackchiaseedwas5.15%of moisture,34.70%offat,4.10%ofash,17.90%ofprotein, 40.25%ofcarbohydrateand17.30%ofrawfiber. Physicochemicalcharacterizationindicatedthepresenceof unsaturatedcompoundsthatarestableforthepresenceof antioxidants. International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(1), 70-72, January (2013)Int. Res. J. Biological Sci.

International Science Congress Association 72 TheFTIRcharacterizationshowedthepresenceof polyunsaturatedcompounds.Finally,theprofileoffattyacids byGC-MSevidencedthepresenceofomega-3fattyacid, vitaminEandphytosterolssuchascampesterol,stigmasterol and -sitosterol; the latter with a concentration of 1233.9 mg/L. References 1.HarveyH.R.,LandpoliticsinthevalleyofMexico:atwo thousandyearperspective,UniversityofNewMexico Press, Albuquerque (1991) 2.CahillJ.P.,Ethnobotanyofchia,SalviahispanicaL. (Lamiaceae), Economic Botany, 57(4), 604-618 (2003) 3.Ayerza R., The Seeds Protein and OilContent, FattyAcid Composition,andGrowingCycleLengthofaSingle Genotypeofchia(SalviahispanicaL.)asAffectedby Environmental Factors, J. Oleo Sci., 58(7), 347-354 (2009) 4.AyerzaR.,Oilcontentandfattyacidcompositionofchia (SalviahispanicaL)fromfivenorthwesternlocationsin Argentina,J.Am.Chem.Soc.,72(9),1079-1081.DOI: 10.1007/BF02660727 (1995) 5.UmhauJ.C.andDauphinaisK.M.,Omega-3 PolyunsaturatedFattyAcidsandHealth,Low-Cost Approaches to Promote Physical and Mental Health, 2, 87-101, DOI: 10.1007/0-387-36899-X_4. (2007) 6.AyerzaR.andCoatesW.,DietaryLevelsofChia: InfluenceonYolkCholesterol,LipidContentandFatty AcidCompositionforTwoStrainsofHens,Poult.Sci., 130(5), 724- 739 (2000) 7.VuksanV.,WhithamD.,SievenpiperJ.L.,JenkinsA.L., RogovikA.L.,BazinetR.P.,VidgenE.andHannaA., SupplementationofConventionalTherapywiththeNovel GrainSalba(SalviahispanicaL.)ImprovesMajorand EmergingCardiovascularRiskFactorsinType2Diabetes, Diabetes Care, 30(11), 2804 -2810 (2007) 8.AwadA.B.andFinkC.S.,Phytosterolsasanticancer dietarycomponents:evidenceandmechanismofaction,J. Nutr., 130(9), 2127-2130 (2000) 9.AlpersD.H.,StensonW.F.,TaylorB.E.andBierD.M. Manualofnutritionaltherapeutics(5thEd.),Philadelphia, PA USA: Lippincot Williams & Wilkins (2008) 10.RatnerR.andOrtzM.,Fitoesteroles:Unaalternativa naturalaltratamientodelahipercolesterolemia,Revista Obesidad, 5(1), 24-29 (2008) 11.BovicP.J.,Theroleofphytosterolsandphytosterolinsin immunemodulation:areviewofthepast10years,Curr. Opin Clin Nutr Metab Care, 4(6), 471-475 (2001) 12.JayarasA.,ToveyFl.andHobsleyM.,DuodenalUlcer prevalence:researchintothenatureofpossibleprotective dietary lipids, Phytother. Res., 17(4), 391-398 (2003) 13.WangT.,HicksK.B.andMorenoR.,Antioxidantactivity of phytosterols, oryzanol, and other phytosterol conjugates,J. Am. Oil Chem. Soc., 79(12), 1201-1206 (2002) 14.Smania E.F., Delle MonacheF. and SmaniaA.,Antifungal activityofsterolsandtriterpenesisolatedfronGanoderma annulare, Fitoteraphy, 74(4), 335-377 (2003) 15.DreikornK.,PhytotherapeuticAgentsintheTreatmentof BenignProstaticHyperplasia,Curr.Urol.Rep.,1(2),103109 (2000) 16.FierroA.,VsquezY.,ReyesP.andBozaS., DeterminacinCuantitativadel-Sitosterolpresenteen vegetalesdeladieta,Posiblesimplicacionesparasuuso preventivoenpoblacionessusceptibles,ClnicaCiencia, 02(02), 43-48 (2004) 17.A.O.A.C.Officialmethodsofanalysis.14th.Ed. AssociationofOfficialAnalyticalChemists.Washington. USA. p. 7-16.(1984) 18.AmericanOilChemistsSociety.PeroxideValue,Acetic Acid-ChloroformMethodCd8-53.Urbana,ILUSA. (2006) 19.American Oil Chemists Society. Iodine Value (Wijs) Cd 1-25. Urbana, IL USA. (2006) 20.CiauS.,RosadoG.,AceretoP.,ChelG.andBentacurD., Propiedades fisicoqumicas y composicin de cidos grasos delaceitedecha(salviahispnicaL),Industria Alimentaria,32-36 (2009) 21.Hesse M., Meier H., Zeeh B., Mtodos Espectroscpicos en Qumica Orgnica, 2da Ed., Sintess, Espaa (1999) 22.SkoogD.A.,HollerF.J.,NiemanT.A.,Principiosde AnlisisInstrumental,5taEd.,McGrawHill,Madrid, Espaa (2001)