c18（ods）で困った時の逆相カラムガイド synergi hydro-rp...

C18ODS

C18ODS

2

C

S

S

F

M

Phenomenex

For the Customer, and for Society. Its our Commitment.For the Customer, and for Society. Its our Commitment.

A3

3

C18

Synergi Fusion-RP

Synergi Fusion-RPC18Fig.1

C18FusionFusion400m2/gFig.1

Columns

DimensionsMobile Phase

Flow RateDetection

Sample

: Synergi 4 Fusion-RP C18 C16+: 150 x 4.6mm: 20mM Potassium Phosphate, pH2.5 / Methanol (70:30): 1.0mL/min: UV @ 210nm: 1. Phenylephrine 2. Phenylpropanolamine 3. Pseudoephedrine

Synergi Fusion-RP(Fig.1)

(Fig.2)

C18 C18

C18C18

min0 1 2 3 4 5

1 2 3

min0 1 2 3 4 5

1 2

3

min0 1 2 3 4 5

1

2 & 3

C10H15NO

2.Phenylpropanolamine 3.Pseudoephedrine

H2NCH3

OH

C9H13NO

CH3

CH3

OHHN

Synergi Fusion

C18

C16+

4

Synergi Fusion-RP

C18FusionFig.1C18Fig.3

LC/MSFusionFig.4

Columns


Flow RateDetection

Sample

: Synergi 4 Fusion-RP C18: 150 x 4.6mm: 20mM Potassium Phosphate, pH2.5 / Acetonitrile (75:25): 1.0mL/min: UV @ 210nm: 1. Maleic acid 2. Chlorpheniramine 3. Triprolidine 4. Diphenhydramine

Columns


Gradient Rate

Flow RateDetection

: Synergi 4 Fusion-RP C18: 150 x 4.6mm: A: 0.1% CH3COOH in Water B: 0.1% CH3COOH in Methanol: 95:5 (A/B) linear to 5:95 over 8 min hold for 5 min.: 0.5 mL/min: Ion Source: ESI Scan Rate: 13000 m/z/s Scan Range: 50-1000

C18Synergi Fusion-RPFig.3

C18Synergi Fusion-RPLC/MSFig.4

1.Maleic acid 4. Diphenhydramine

OH

OHO

O

IN

O

min0 2 4 6 8 10 12 14

1

2 34

min2 4 6 8 10 12 14

1

2

3

4

0 1 2 3 4 5 6 7 8 90 1 2 3 4 5 6 7 8 9 min

0.1

x106

0.2

0.3

0.4

0.5

0.6

x107

TIC

: Tot

al Io

n C

urre

nt

Synergi Fusion

C18

A3

5

C18

Synergi Hydro-RP

Synergi Hydro-RPC18Fig.5

100%Fig.6C18100%Fig.7

Column

DimensionsOrder NO.

Mobile PhaseFlow Rate

TemperatureDetection

Sample

: Synergi 4 Hydro-RP: 250 x 4.6mm: 00G-4375-E0: 20mM Potassium Phosphate, pH2.9: 0.7mL/min: 22C: UV @ 220nm: 1. Oxalic acid 2. Tartaric acid 3. Glycolic acid 4. Formic acid 5. Pyruvic acid 6. Malonic acid 7. Acetic acid 8. Maleic acid 9. Citric acid

Synergi Hydro-RP(Fig.5)

Synergi Hydro-RP (Fig.6)

min

min

n

n

6

Synergi Hydro-RP

C18

1.AtenololC18Synergi Hydro-RPFig.8


GradientFlow Rate

TemperatureDetectionInjectionSample

Mobile phaseSample

: 20mM Potassium phosphate: Nucleotides

: 150 x 4.6mm: A=10mM Triethylammonium formate pH 6.0 B=Acetonitrile with 10mM Triethylammonium formate : A/B (85:15) to A/B (35:65) in 15 minutes: 1.5mL/min: Ambient: UV @ 230nm: 1L of beta-blockers mix each at 0.8g/L : 1. Atenolol 2. Pindolol 3. Nadolol 4. Acebutolol 5. Metoprolol 6. Labetalol

100% (Fig.7)

Synergi Hydro-RP (Fig.8)

1.Atenolol

HNO

OCH3

CH3H2N

OH

C18 Synergi Hydro-RP

C18

Synergi Hydro-RP

02 4 6 8 10 12

Day 1

02 4 6 8 10 12

Day 1

02 4 6 8 10 12 14 min

Day 2

02 4 6 8 10 12

Day 2

A3

7

Synergi Max-RP

Synergi Max-RPC12C18Fig.9Fig.10

C18Fig.11Synergi Max-RPFig.11


Gradient

Flow RateDetectionInjection

TemperatureSample

: 50 x 4.6mm : A= Water with 0.1% Formic acid B= Acetonitrile with 0.1% Formic acid: A/B (95:5) to 100% B in 5min: 1.5mL/min:UV @ 254nm: 5L: 30C: 1. Thiourea (0.25g), 2. Codeine (1g), 3. Chlorpheniramine (2g), 4. Propranolol (6g), 5. Desipramine (0.5g), 6. Ibuprofen (6g),

Synergi Max-RP(Fig.9) Synergi Max-RP(Fig.10)

Synergi Max-PR (Fig.11)

4.Propranolol

C18

Synergi Max

C12

C12 C12

C12

C12

C12 C12

NHO

OH

8

Synergi Max-RP

TFA

Synergi Max-RPC12Fig.13

Synergi Max-RPLC/MSTFAFig.12

Column

Mobile phaseFlow rate

Column oven temperatureInjection volume

Probe voltageNebulizer gas flow

Drying gas pressureProbe temperature

CDL temperatureBlock heater temperature

CDL & Q-array voltageInterval

SIM

: Phenomenex Synergi MAX-RP (2.0 mm I.D. 150 mmL.): 0.1% formic acid-water / acetonitrile (70:30): 0.3 mL/min: 40 C: 2 L:+4.5 kV (APCI-Positive mode): 2.5 L/min: 0.04 MPa : 350 C: 200 C : 200 C : using default values : 0.8 sec / 16 chs: m/z 402.1,361.1,343.1,325.1 for prednisolone m/z434.1, 393.1, 373.1, 355.1 for betamethasone & dexamethasone m/z 476.1, 435.1, 415.1 for triamcinolone acetonide m/z 494.1, 453.1, 413.1 for fluocinolone acetonide m/z 446.1, 405.1 for hydrocortisone acetate

ColumnDimensions

Order No.Mobile Phase

GradientFlow RateDetectionInjection

TemperatureSample

: Synergi 4 Max-RP: 50 x 4.6mm: 00B-4337-E0: A= Water with 0.1% TFA, Formic acid, or Acetic acid B = Acetonitrile with 0.1% TFA, Formic acid, or Acetic acid: A/B (95:5) to 100% B in 5min: 1.5mL/min: UV @ 254 nm: 5L: 30C: 1. Thiourea (0.25g), 2. Codeine (1g), 3. Chlorpheniramine (2g), 4. Propranolol (6g), 5. Desipramine (0.5g), 6. Ibuprofen (6g),

TFA (Fig.12)

Synergi Max-RP6SIM(Fig.13)

0.1% Formic acid

0.1% Acetic acid

0.1% TFA

0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 16.0 17.0 18.0 19.0 20.0

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1.0

1.1

1.2

1.3

1.4

1.5

1.6

1.7

1.8

(1,000,000)

405.10 (1.00)446.15 (1.00)413.10 (1.00)453.10 (1.00)494.10 (1.00)415.10 (1.00)435.10 (1.00)476.10 (1.00)355.05 (1.00)373.10 (1.00)393.10 (1.00)434.10 (1.00)325.00 (1.00)343.05 (1.00)361.10 (1.00)402.15 (1.00)

q w

e

r

t

y

1: Prednisolone

2: Dexamethasone3: Betamethasone

4: Triamcinolone acetonide

5: Fluocinolone acetonide

6: Hydrocortisone acetate

O

CH3

HOCH3

CH2OHO

OHCH3

F

BetamethasoneC22H29FO5

Exact Mass: 392.20Mol. Wt.: 392.46

O

CH3

HOCH3

CH2OHO

OHCH3

F

DexamethasoneC22H29FO5

Exact Mass: 392.20Mol. Wt.: 392.46

No.C43A

A3

9

Synergi Polar-RP

Synergi Polar-RPFig.14

C8C18Fig.6PolarFig.15

Columns

Mobile phaseFlow rate

Components

: Luna 5m C8(2) 150x4.6mm Synergi 4m Polar-RP150x4.6mm: 65:35 10mM Ammonium acetate pH 3.5:Methanol: 1 mL/min: 1. Cephradine 2. Cefadroxil 3. Cephalexin 4. Cefaclor 5. Cefazolin

0)

sone

Synergi Polar-RP(Fig.14)

Synergi Polar-RP(Fig.15)

2.Cefadroxil 3.Cephalexin

NH

S

O

O

H H

HO

NH2CH3

COOH

N

NH

S

O

O

H H

NH2CH3

COOH

N

Luna C8(2)

min0 1 2 3 4 5 6

0

20

40

60

80

100

120

140

160

1

2

3

4

5

Synergi Polar-RP

min0 1 2 3 4 5 6

0

25

50

75

100

125

150

175

1

5

2-4

10

Synergi Polar-RP

part2

100%Fig.17


Flow RateDetection

TemperatureSample

: 150 x 4.6mm : 20mM Potassium phosphate pH2.5/ Methanol (97:3) : 1.0mL/min : UV @ 220nm : 25C: 1. Formic acid 2. Acetic acid


Flow RateDetectionInjection

TemperatureSample

: 150 x 4.6mm: 20mM Potassium phosphate pH 3 /Methanol (50:50): 1.0mL/min: UV @ 230nm: 2L: 22C: 1. Metaproterenol (0.4g), 2. Pindolol (0.6g), 3. Metoprolol (0.15g), 4. Alprenolol (0.3g), 5. Propranolol (0.04g), 6. Ethylparaben (0.4g),

Synergi Polar-RPpart2(Fig.16)

Synergi Polar-RP100%(Fig.17)

1. Metaproterenol

1. Formic acid 2. Acetic acid

2.Pindolol

3.Metoprolol

H

O

OHC

HO

OH

OH

HN CH3

CH3

OH

HNO

HN

CH3

CH3H3CO

OH

H

HNO

O

OHC

Synergi Polar

C18

Synergi Polar

C18

C16+

C18

C18

A3

11

o-,m-,p-

Luna PFP2

Luna PFP2Fig.18Fig.19Fig.20

C18Fig.19

ColumnDimension

Part No.Mobile Phase

Flow RateTemperature

DetectionSample

: 16298Luna 3 m PFP(2): 150 x 4.6 mm: 00F-4447-E0: Water/ Methanol (50:50): 1 mL/min: 22 C: UV @ 254 nm: 1. o-Methylacetophenone o- 2. p-Methylacetophenone p- 3. m-Methylacetophenone m-

ColumnDimension



DetectionColumn

: Luna 3 m C18(2): 150 x 4.6 mm: 00F-4251-E0: Water/ Methanol (50:50): 1 mL/min: 22 C: UV @ 254 nm: 1. p-Methylacetophenone p- 2. o-Methylacetophenone o- 3. m-Methylacetophenone m-

ColumnDimensionPart No.Mobile Phase

GradientFlow RateTemperatureDetectionSample

: Luna 3 m PFP(2): 150 x 4.6 mm: 00F-4447-E0: A: 0.1 % Formic acid in Water B: 0.1 % Formic acid in Acetonitrile: A/B (60:40) to (50:50) in 10 min: 1 mL/min: 22 C: UV @ 254 nm: 1.2,3-Dimethylphenol 6.2,5-Dichlorophenol 2.2,5-Dimethylphenol 7.2,6-Dichlorophenol 3.2,6-Dimethylphenol 8.3,4-Dichlorophenol 4.3,4-Dimethylphenol 9.3,5-Dichlorophenol 5.3,5-Dimethylphenol 10.2,4-Dibromophenol

Luna PFP2(Fig.18)

LunaPFP2o-,m-,p (Fig.19)

3,4-,3,5-Dichloro-,Dibromo-LunaPFP2 (Fig.20)

Positional Isomers of Methylacetophenone on Luna 3 m PFP(2) App ID 16298 Positional Isomers of Methylacetophenone on Luna 3 m C18(2) App ID 16299

0 2 4 6 8 10 12 14 16 18 min

1

2

3

0 2 4 6 8 10 12 14 16 18

1

2

3

min

F

F

F

F

F

1234

5

67

8

9

10

0 2 4 6 8 10 12 14 min

16297

0 2 4 6 8 10 12 14 min

1 234

5

67

8

9 & 10

olar

Luna PFP2 C18

12

Luna Phenyl-Hexyl-

-

-!

J

--

Luna Phenyl-Hexyl

Ju

P

ODS

ODS

Phenyl-Hexyl

1+2 3

4 56

12

3

4 56

C8

C18

Luna 5 m Phenyl-Hexyl, 150x4.6mm Luna 5 m C8(2), 150x4.6mm Luna 5 m C18(2), 150x4.6mm: 50:15:35 Methanol:Acetonitrile: 20mM Ammonium Formate pH 3.5: 1.0 mL/min: 1. Rhodamine 123 2. Rhodamine B Base 3. Rhodamine 6G

: ODS1504.65 Luna Phenyl-Hexyl1504.65: 50:50 Acetonitrile: 20mM Potassium phosphate pH 2.5: 1.0 mL/min: 1. Indoprofen 2. Ethyl paraben 3. Naproxen 4. Diflunisal 5. Indomethacin 6. Ibuprofen

Luna Phenyl-Hexyl

-

R

Rhodamine123

6.Ibupurofe

1.Indoprofen 2.Ethyl paraben

5.Indomethacin

A3

13

Jupiter Proteo 90

C12ODS

Jupiter Proteo

Jupiter Proteo

ColumnsDimensions


Gradient


DetectionSample

: Jupiter 4 m Proteo 90 : 250 x 4.6 mm: 00G-4396-E0: A) 0.012 % TFA in Water B) 0.01 % TFA in Acetonitrile: A/B (95:5) for 5 min, then to A/B (60:40) in 55 minutes: 1 mL/min: 22C: UV @ 210 nm: Top Chromatogram -Lactoglobulin tryptic digest Bottom Chromatogram Oxidized -Lactoglobulin tryptic digest

ColumnsDimensions


Gradient


DetectionSample

: Jupiter 4 m Proteo 90 : 250 x 4.6 mm: 00G-4396-E0: A) 0.012 % TFA in Water B) 0.01 % TFA in Acetonitrile: A/B (95:5) for 5 min, then to A/B (60:40) in 55 min: 1 mL/min: 22 C: UV @ 210 nm: Top Chromatogram RNase tryptic digestBottom Chromatogram Deamidated RNase tryptic digest

1Fig.21

2Fig.22

App

ID 1

4394

App

ID 1

4395

App

ID 1

4392

App

ID 1

4393

-

RNase

RNase

-

14

Jupiter Proteo 90 J

Jupiter Proteo10kDa125

Ju

Columns

DimensionsPart No.

Mobile Phase

Gradient


DetectionSample

: Jupiter 4 m Proteo 90 AODS BODS CODS: 250 x 4.6 mm: 00G-4167-E0: A) 0.1 % TFA in Water B) 0.085 % TFA in Acetonitrile: A/B (95:5) to A/B (55:45) in 20 minutes: 1 mL/min: 22 C: UV @ 214 nm: 1. NH2-Arg-Gly-Gly-Ala-Gly-Gly-Leu- Gly-Leu-Gly-Lys-Amide 2. Ac-Arg-Gly-Gly-Gly-Gly-Gly-Leu-Gly- Leu-Gly-Lys-Amide 3. Ac-Arg-Gly-Ala-Gly-Gly-Gly-Leu-Gly- Leu-Gly-Lys-Amide 4. Ac-Arg-Gly-Val-Gly-Gly-Gly-Leu-Gly- Leu-Gly-Lys-Amide 5. Ac-Arg-Gly-Val-Val-Gly-Gly-Leu-Gly- Leu-Gly-Lys-Amide

ColumnsDimensions


GradientFlow RateDetectionSample

: Jupiter 4 m Proteo 90 : 250 x 4.6 mm: 00G-4396-E0: A) 0.1 % TFA in Water B) 0.085 % TFA in 95:5 Acetonitrile/Water: A/B (80:20) to A/B (20:80) in 15 minutes: 1 mL/min: UV @ 220 nm: Human Insulin

Fig.23

Fig.24

J

J

App

ID 1

4914

min2 4 6 8 10

mAU

0

200

15 min @ 60 C

400

App

ID 1

4917

14917

min2 4 6 8 10

mAU

0

200 60 min @ 60 C

App

ID 1

4915

mAU

200

0

min2 4 6 8 10

30 min @ 60 C

App

ID 1

4428

App

ID 1

4427

App

ID 1

4426

App

ID 1

4429

12

3 4 5

Jupiter 4 m Proteo 90

AODS

BODS

CODS

A3

15

HPLC

Jupiter 300

10kDa300C4C5C18

C18Jupiter 300C4

ColumnsDimensions


Gradient


DetectionSample

: Jupiter 300 5 m C18 300 : 250 x 4.6 mm: 00G-4053-E0: A) 0.1 % TFA in Water B) 0.1 % TFA in Acetonitrile: A/B (75:25) to A/B (45:55) in 15 min (2 % B/min): 1 mL/min: Ambien: UV @ 220 nm: 1. Equine Cytochrome c 2. Bovine Cytochrome c 3. Canine Cytochrome c

ColumnsDimensions


Gradient


DetectionInjection Volume

Sample

: Jupiter 300 5 m C4 300 : 150 x 4.6 mm: 00F-4167-E0: A) 0.1 % TFA in Water B) 0.08 % TFA in Acetonitrile: A/B: (95:5) to A/B: (20:80) in 20 minutes: 1 mL/min: 22 C: UV @ 280 nm: 25 L: 1. Bovine Serum Albumin 2. Glutamic Dehydrogenase 3. -Galactosidase 4. Ovalbumin

Jupiter 300C18Fig.25

Jupiter 300C4Fig.26

App

ID 8

715

20 4 6 8 10 12 14 min

App

ID 1

4761

1

2

3

4

16

Clarity Oligo-RP & Clarity Oligo-WAX

G

ClarityHPLCDNARNA

TwinpH112 60

GC

N-1 impurities

p

ColumnsDimensions


Gradient

Flow RateDetection

Sample

: Clarity 3 m Oligo-RP C18: 50 x 4.6 mm: 00B-4441-E0: A: 10 mM TPAC B: Methanol: A/B (75:25) to A/B (55:45) in 80 min (50 min run time): 1 mL/min: UV @ 260 nm: 1. 20 nt RNA with sequence CUGUAAUCUCUUGUCUATT (2.5g) 2. 21 nt RNA with sequence UCUGUAAUCUCUUGUCUATT (2.5g)

ColumnsDimensions


GradientFlow RateDetection

TemperatureSample

: Clarity 3 m Oligo-RP: 50 x 4.6 mm ID: 00B-4441-E0: A: 50 mM TEAA / 5 % Acetonitrile B: Methanol: A/B (90:10) to A/B (60:40) in 20 min : 1mL/min: UV @ 260 nm: ambient: 1. FAM-DNA (25 nt) GC% 72 % 2. FAM Labeled DNA (25 nt) GC% 56 % 3. FAM Labeled DNA (25 nt) GC% 20 %

ColumnsDimensions


GradientFlow RateDetection

TemperatureSample

: Clarity 10 m Oligo-WAX: 150 x 4.6 mm ID: 00F-4451-E0: A: 20 mM Tris pH 8.0 / 10 % Acetonitrile B: 20 mM Tris pH 8.0 / 10 % Acetonitrile / 1.2 M NaCl: 100 % A to 100 % B in 20 min: 2.2 mL/min: UV-Vis Abs. - Diode Array 260 nm: ambient: 1. Desalted DNA 20 nt

Clarity Oligo-RPFig.27

3

Clarity Oligo-RP 3Fig.28

360

Clarity Oligo-WAXFig.29

min0 5 10 15 20 25 30 35 40

mAU

0

10

20

30

40

50

21 nt20 nt

App

ID 1

5935

App

ID 1

5990

12

3

0 10 20 min

0

100

200

300

mAU

App

ID 1

6395

100

0

20

40

60

80

100

120

140

mAU

20 min

20 nt

1

A3

0%

20%

40%

60%

80%

100%

120%

0 100 200 300 400 500 600

Column A

Column B

Column C

Column D

Column E

Column F

Gemini-NX

Time of exposure (hour)

Gemini-NX

Per

cent

Effi

cien

cy

Si

Si

OH- OH-

OH-H+

H+H+

CH2CH2

O

O

O

Si

Si

O

OO

O

17

pHODS

Gemini-NX

Si-CH2-CH2-Si23

Multi Functional

pH112

pH10.5

/

Fig.30

18

Gemini-NX

Gemini-NX 5m C18 A 5m C18

Fig.31

-Fig.32

0 10 20 min

1

2

3

4,56

78

9

10A

pp ID

166

42

0 10 20 min

1

2

3

4 6

7

8

9

10

}

5

}

App

ID 1

6641

Conditions for all columnsDimensions

Mobile PhaseGradient


Detection

: 150 x 4.6 mm: A: 10 mM Ammonium Bicarbonate pH 10.5 B: Acetonitrile: A/B (85:15) to (70:30) in 15 min to (50:50) in 5 min, Hold for 5 min: 1.5 mL/min: Ambient: UV @ 230 nm

Sample : 1.Bisoprolol Contaminant 2.Sotalol 3.Atenolol 4.Labetolol (Diastereoisomeric Pair) 5.Nadolol (Diastereoisomeric Pair) 6.Pindolol 7.Metoprolol 8.Bisoprolol 9.Propranolol 10.Alprenalol

5.Nadolol

H3CCH3

CH3O

OH

OH

HO

NH

HN

O

CH3

H2N

HO

HO4.Labetalol

A3

4mm

19

SecurityGuard

1

2 3

1 2

SecurityGuard SecurityGuard

SecurityGuard SecurityGuard

3

HPLC

4 350

22

HPLCC18

HPLC

C18ODS

111- 00535 - 20 - 8 CS5 FTEL : 03 -5835 - 0120 FAX : 03 -5835 - 0124

533- 0033 1 - 18 - 22 9 FTEL : 06 -6328 -2255 FAX : 06 -6328 -2277

[email protected]://solutions.shimadzu.co.jp/glc

c18（ods）で困った時の 逆相カラムガイド synergi hydro-rp...

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c18（ods）で困った時の逆相カラムガイド synergi hydro-rp...