Selection guide of mobile phase · (1) The effect of organic solvent type and content - In general,...

Selection guide of mobile phase COMOSIL HILIC column generates retention and separation by hydrophilic interaction (mainly hydrogen

bond) and anion-exchange. Refer to following recommendations to select an appropriate mobile phase condition. (1) The effect of organic solvent type and content - In general, acetonitrile/water is used as mobile phase. - Retention increases as water content in the mobile phase decreased. (Fig.1) - Use acetonitrile content in the mobile phase within the range of 0-95% (in general 50-95%). - Methanol/water generates shorter retention than acetonitrile/water. (Fig.2) - Use only HPLC grade solvents.

012345678

65 70 75 80 85 90 95 100

Acetonitrile content(%)

k'

meso-Erythritol

Leucine

Tris(hydroxymethyl)aminomethane

Guanidoacetic acid

012345678

Acetonitorile Methanol

k'

meso-Erythritol

Leucine


Fig.1 The effect of acetonitrile content on retention Fig.2 Difference of acetonitrile and methanol on retention

Column; COSMOSIL HILIC Column; COSMOSIL HILIC

Mobile phase; Acetonitrile/ 10mmol/l CH3COONH4 Mobile phase; Organic solvent/ 10mmol/l CH3COONH4 = 90/10

(2) The effect of buffer pH - Keep pH of the mobile phase within the range of 2-7.5.

- The buffer around neutrality generates larger retention. (Fig.3) Fig.3 The effect of buffer pH on retention Column; COSMOSIL HILIC

Mobile phase; Acetonitrile / 10mmol/l buffer = 90/10

(3) The effect of salt type and concentration - When analyze ionic compounds, add salts or buffers in the mobile phase. - When mobile phase has high acetonitrile content, note dissolubility of the salt. The dissolubility of

phosphate buffers used widely in reversed phase chromatography is low in acetonitrile. Therefore use of phosphate buffers is not recommended. Keep the concentration of acetonitrile under 70% if use a phosphate buffer. Check that the salt does not precipitate when mixed with acetonitril before use.

- Ammonium acetate or formic acid ammonium buffers are recommended because they are soluble in high t acetonitrile content.

0

1

2

3

4

5

2 3 4 5 6 7 8 9

pH

k'

meso-Erythritol

Leucine


Guanidoacetic Acid

- Use the buffer concentration within the range of 5 - 100mmol/l. Moreover, Check that the salt does not precipitate after mixing buffer and acetonitrile.

- High salt concentration inhibits ion exchange and decreases retention. (Fig.4) - Run mobile phase through membrane filter (less than 0.45µm in pore size) before use.

Fig.4 The effect of salt concentration on retention Column: COSMOSIL HILIC

Mobile phase; Acetonitrile / 10mmol/l CH3COONH4 = 50/50

(4) Selection of mobile phase

Following are the recommended mobile phases for different compound types.

Neutral compounds Acetonitrile / Water = 90/10 Basic compounds Acetonitrile / 10mmol/l CH3COONH4 = 90/10 Amphoteric compounds Acetonitrile / 10mmol/l CH3COONH4 = 70/30 Acidic compounds Acetonitrile / 10mmol/l CH3COONH4 = 50/50

not eluted Acetonitrile / 10mmol/l Phosphate buffer (pH7.0)= 50/50 (5) Improvement of peak shape

Try following if peak shape is tailing. The peak shape might improve. - Add 5mmol/l EDTA to mobile phase. - Change to citrate buffer. (i. e. 10 mmol/l citrate buffer pH7.0)

80%Acetonitrile/ 80%Acetonitrile/

10mmol/l CH3COONH4 10mmol/l CH3COONH4+ 5mmol/l EDTA

Fig.5 Improvement of peak shape

Column: COSMOSIL HILIC(4.6mmI.D.-250mm)

Sample: Tryptophan(1ng), Flow rate: 1.0ml/min

Detection: UV254nm, Temperature: 30°C

(6) Others - Use scrupulously degassed mobile phase. Air bubbles generate detection noise and accelerate column

deterioration.

- We recommend keeping the chromatography conditions constant, since frequent changes of mobile phase shorten column life.

0

2

4

6

8

10

0 25 50 75 100

Buffrer concentration(mmol/l)

k' Benzoic acid

IndexSample name Cas No. Page Sample name Cas No. Page

A Acetamide 60-35-5 1 L‐Homoserine 672-15-1 14

Acetazolamide 59-66-5 1 Hydantoic Acid 462-60-2 14

Acetrizoic Acid 85-36-9 1 Hydantoin 461-72-3 14

Acrylic Acid 79-10-7 1 Hydroxylamine‐O‐sulfonic Acid 2950-43-8 14

L-α-Alanine 56-41-7 1 cis-4-Hydroxy-D-proline 2584-71-6 14

β‐Alanine 107-95-9 1 L‐Hydroxyproline 51-35-4 15

Allantoic Acid 99-16-1 2 N‐Hydroxysuccinimide 6066-82-6 15

Allantoin 97-59-6 2 I Indigo carmine 860-22-0 15

p‐Aminobenzamidine 3858-83-1 2 D-Isoascorbic Acid 89-65-6 15

p‐Aminobenzoic Acid 150-13-0 2 Isocinchomeronic Acid 100-26-5 15

4‐Amino‐n‐butyric Acid 〔GABA〕 56-12-2 2 L-Isoleucine 73-32-5 15

6‐Aminohexanoic Acid 60-32-2 2 Isonicotinic Acid 55-22-1 16

5‐Aminolevulinic Acid 106-60-5 3 Isonicotinohydrazide 54-85-3 16

2-Aminopyridine 504-29-0 3 Isopropyl β-D-1-thiogalactopyranoside (IPTG) 367-93-1 16

3-Aminopyridine 462-08-8 3 K Kojic Acid 501-30-4 16

5-Amino-1H-tetrazole 4418-61-5 3 L L-Leucine 61-90-5 16

3‐Amino‐1H‐1,2,4‐triazole 61-82-5 3 N-D-Leucyl-L-tyrosine 3303-29-5 16

5-Aminouracil 932-52-5 3 L‐Lysine 56-87-1 17

Amphotericin B 1397-89-3 4 M Maleic Acid 110-16-7 17

Angiotensin I(Human) 484-42-4 4 L‐(-)‐Malic Acid 97-67-6 17

Angiotensin II(Human) 4474-91-3 4 Malonic Acid 141-82-2 17

L‐Arginine 74-79-3 4 Mecobalamin 13422-55-4 17

L(+)‐Ascorbic Acid 50-81-7 4 Metanilic Acid 121-47-1 17

L-Asparagine 70-47-3 4 L‐Methionine 63-68-3 18

L‐Aspartic Acid 56-84-8 5 6-Methyl-2-thiouracil 56-04-2 18

6‐Azauracil 461-89-2 5 N‐Methylglucamine 6284-40-8 18

Aztreonam 78110-38-0 5 N‐Methylhydroxylamine 593-77-1 18

B Benzamidine 618-39-3 5 Mucic Acid 526-99-8 18

Benzenesulfonic Acid 98-11-3 5 Murexide 3051-09-0 18

Benzoic Acid 65-85-0 5 N Nicotinamide 98-92-0 19

Bromoacetic Acid 79-08-3 6 Nicotinic Acid 59-67-6 19

C Cacotheline 561-20-6 6 L‐Noradrenaline 51-41-2 19

Camostat 59721-28-7 6 DL-Norleucine 616-06-8 19

L‐Carnitine 541-15-1 6 DL‐Norvaline 760-78-1 19

Ceftriaxone 73384-59-5 6 O L‐Ornithine 70-26-8 19

Chloroacetic Acid 79-11-8 6 Orotic Acid 65-86-1 20

Citrazinic Acid 99-11-6 7 Oxalic Acid 144-62-7 20

Creatine 57-00-1 7 Oxamic Acid 471-47-6 20

Creatinine 60-27-5 7 Oxytocin 50-56-6 20

Cyanoacetic Acid 372-09-8 7 P D‐Pantothenic Acid 79-83-4 20

Cyanuric Acid 108-80-5 7 L-(-)-Phenylalanine 63-91-2 20

L‐Cysteine 52-90-4 7 p‐Phenylenediamine 106-50-3 21

L‐(-)‐Cystine 56-89-3 8 L‐(+)‐α‐Phenylglycine 2935-35-5 21

Cytidine 65-46-3 8 Phosphocreatine 67-07-2 21

Cytosine 71-30-7 8 O‐Phospho‐L‐serine 407-41-0 21

D 3,4-Diaminobenzoic Acid 619-05-6 8 Picolinic acid 98-98-6 21

3,5-Diaminobenzoic Acid 535-87-5 8 Pivalic Acid 75-98-9 21

2,4-Diaminophenol 95-86-3 8 Procaterol 72332-33-3 22

DL‐2,6‐Diaminopimelic Acid 583-93-7 9 L-Proline 147-85-3 22

DL‐2,3‐Diaminopropionic Acid 54897-59-5 9 Propionic Acid 79-09-4 22

Diatrizoic Acid 117-96-4 9 Pyruvic Acid 127-17-3 22

Dipicolinic acid 499-83-2 9 R Ribose‐5‐phosphate 4300-28-1 22

Dithiouracil 2001-93-6 9 S D‐Saccharic Acid 87-73-0 22

L‐DOPA 59-92-7 9 Sarcosine 107-97-1 23

Dopamine 51-61-6 10 Sebacic Acid 111-20-6 23

E meso‐Erythritol 149-32-6 10 L-Serine 56-45-1 23

F Famotidin 76824-35-6 10 Sinigrin 3952-98-5 23

Folic Acid 59-30-3 10 Succinic Acid 110-15-6 23

Folinic Acid 58-05-9 10 Sulbactam 68373-14-8 23

Formamide 75-12-7 10 Sulfanilic acid 121-57-3 24

D‐Fructose‐6‐phosphate 643-13-0 11 T L‐(+)‐Tartaric Acid 87-69-4 24

Fuchsine, Acid 〔Rubin S〕 3244-88-0 11 Taurine 107-35-7 24

Fumaric Acid 110-17-8 11 L-Theanine 3081-61-6 24

G Gluconic Acid 526-95-4 11 2‐Thiobarbituric Acid 504-17-6 24

α‐D‐Glucose‐1‐phosphate 59-56-3 11 2‐Thiouracil 141-90-2 24

D‐Glucose‐6‐phosphate 56-73-5 11 L-Threonine 72-19-5 25

D-Glucuronic Acid 6556-12-3 12 Todralazine 14679-73-3 25

L‐Glutamic Acid 56-86-0 12 Trichloroacetic Acid 76-03-9 25

L‐Glutamine 56-85-9 12 Tris(hydroxymethyl)aminomethane 77-86-1 25

Glutaric Acid 110-94-1 12 L-Tryptophan 73-22-3 25

DL‐Glyceric Acid 600-19-1 12 L-Tyrosine 60-18-4 25

Glycinamide 598-41-4 12 U Uracil 66-22-8 26

Glycine 56-40-6 13 Urea 57-13-6 26

Glycolic Acid 79-14-1 13 Uridine 58-96-8 26

Glycylglycine 556-50-3 13 V L-Valine 72-18-4 26

Guanidoacetic Acid 352-97-6 13

H 1,2,6-Hexanetriol 106-69-4 13

L-Histidine 71-00-1 13

L‐Homocystine 626-72-2 14

1

COSMOSIL Chromatogram IndexCOSMOSIL Chromatogram Index

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Sample: AcetamideCAS No.: [60-35-5]Molecular formula: C2H5NOColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ H2O=95/5Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV220 nmAttenuation: 0.128 aufsSample conc.: 10.0mg/mlInjection volume: 0.5µlRetention time: 4.75minCapacity factor: 0.57

O

NH2

CH3


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Sample: AcetazolamideCAS No.: [59-66-5]Molecular formula: C4H6N4O3S2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Ammonium acetate =90/10 Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV254 nmAttenuation: 0.128 aufsSample conc.: 0.2mg/mlInjection volume: 0.5µlRetention time: 5.99minCapacity factor: 1.05

S OO

H2N

S NN

HN

OCH3


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Sample: Acetrizoic AcidCAS No.: [85-36-9]Molecular formula: C9H6I3NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Ammonium acetate =50/50Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV254 nmAttenuation: 0.128 aufsSample conc.: 0.8mg/mlInjection volume: 1.0µlRetention time: 16.39minCapacity factor: 4.76

I I

O OH

HN

IO CH3


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Sample: Acrylic AcidCAS No.: [79-10-7]Molecular formula: C3H4O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HOCH2


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Sample: L-α-AlanineCAS No.: [56-41-7]Molecular formula: C3H7NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Ammonium acetate=70/30Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV 210nmAttenuation: 0.128 aufsSample conc.: 5.0mg/mlInjection volume: 2.0µlRetention time: 6.67minCapacity factor: 153

NH2

H3C

H

O

OH


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Sample: β‐AlanineCAS No.: [107-95-9]Molecular formula: C3H7NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Ammonium acetate=70/30Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV 210nmAttenuation: 0.128 aufsSample conc.: 10.0mg/mlInjection volume: 0.5µlRetention time: 7.38minCapacity factor: 1.81

O

HO NH2

2


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Sample: Allantoic AcidCAS No.: [99-16-1]Molecular formula: C4H8N4O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN

HN

O

OH

O

NH2O

H2N


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Sample: AllantoinCAS No.: [97-59-6]Molecular formula: C4H6N4O3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH

HN

HN

O

O

O

NH2


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Sample: p‐AminobenzamidineCAS No.: [3858-83-1]Molecular formula: C7H9N3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Citrate buffer(pH7.0)=70/30 Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV254 nmAttenuation: 0.128 aufsSample conc.: 0.5mg/mlInjection volume: 1.0µlRetention time: 6.07minCapacity factor: 1.31

H2N NH

NH2


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Sample: p‐Aminobenzoic AcidCAS No.: [150-13-0]Molecular formula: C7H7NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O OH

NH2


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Sample: 4‐Amino‐n‐butyric Acid CAS No.: [56-12-2]Molecular formula: C4H9NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Ammonium acetate=70/30Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV210 nmAttenuation: 0.128 aufsSample conc.: 10.0mg/mlInjection volume: 1.0µlRetention time: 7.67minCapacity factor: 1.92

O

HONH2


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Sample: 6‐Aminohexanoic AcidCAS No.: [60-32-2]Molecular formula: C6H13NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HONH2

3


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Sample: 5‐Aminolevulinic AcidCAS No.: [5451-09-2]Molecular formula: C5H9NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

H2N

O

OH


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Sample: 2-AminopyridineCAS No.: [504-29-0]Molecular formula: C5H6N2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


N

NH2


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Sample: 3-AminopyridineCAS No.: [462-08-8]Molecular formula: C5H6N2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2

N


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Sample: 5-Amino-1H-tetrazoleCAS No.: [4418-61-5]Molecular formula: CH3N5Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NHN

NH2

NN


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Sample: 3‐Amino‐1H‐1,2,4‐triazoleCAS No.: [61-82-5]Molecular formula: C2H4N4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NN

NH2

HN


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Sample: 5-AminouracilCAS No.: [932-52-5]Molecular formula: C4H5N3O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH

OH2N

NH

O

4


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Sample: Amphotericin BCAS No.: [1397-89-3]Molecular formula: C47H73NO17Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


CH3O

O

OH OH OH O

H3C

HO

H3C

OHOH

COOHH

OH

OH

O CH3OHNH2

O

OH


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Sample: Angiotensin I(Human)CAS No.: [484-42-4]Molecular formula: C62H89N17O14Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN H

H3C H

NH

O

H3C

O

H

NH

H

N

O

N

O

H

HN

HNH

HN

O

CH3

H3C

OH

O

H

NH

H

HN

O

O

H

NH2

HHN

HN

O

N

O

HONH2

NH

H

HN

OH

OCH3

H3C


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Sample: Angiotensin II(Human)CAS No.: [4474-91-3]Molecular formula: C50H71N13O12Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


N

H

HN

OO

NHH

H

HO

ON

O

NH

HNH

CH3

H CH3

OHN H

OHN

H

CH3

H3C

HO

OHN

H

O

NH2

H

HN

O

HO

NH2HN


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Sample: L‐ArginineCAS No.: [74-79-3]Molecular formula: C6H14N4O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Citrate buffer(pH7.0)=60/40Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV210 nmAttenuation: 0.128 aufsSample conc.: 10.0mg/mlInjection volume: 1.0µlRetention time: 7.97minCapacity factor: 1.95

NH2HNH

H2N

NH

OH

O


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Sample: L(+)‐Ascorbic AcidCAS No.: [50-81-7]Molecular formula: C6H8O6Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Ammonium acetate=50/50Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV245nmAttenuation: 0.128 aufsSample conc.: 0.2mg/mlInjection volume: 3.0µlRetention time: 17.80minCapacity factor: 5.31

O

HOH

H

HO

HO

HO O


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Sample: L-AsparagineCAS No.: [70-47-3]Molecular formula: C4H8N2O3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2HO

OH

O

NH2

5


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Sample: L‐Aspartic AcidCAS No.: [56-84-8]Molecular formula: C4H7NO4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2HO

HOOH

O


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Sample: 6‐AzauracilCAS No.: [461-89-2 ]Molecular formula: C3H3N3O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN

NH

ON

O


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Sample: Aztreonam CAS No.: [78110-38-0]Molecular formula: C13H17N5O8S2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 20mmol/l

Phosphate buffer(pH7)=50/50 Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV280 nmAttenuation: 0.128 aufsSample conc.: 2.5mg/mlInjection volume: 1.0µlRetention time: 17.57minCapacity factor: 5.18

NHH

N

CH3

H

OS

O

HO

OO

N

N

O

S

H2N

CH3

CH3

O

OH


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Sample: BenzamidineCAS No.: [618-39-3]Molecular formula: C7H8N2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


H2N NH


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Sample: Benzenesulfonic AcidCAS No.: [98-11-3]Molecular formula: C6H6O3SColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


S OHOO


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Sample: Benzoic AcidCAS No.: [65-85-0]Molecular formula: C7H6O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O OH

6


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Sample: Bromoacetic AcidCAS No.: [79-08-3]Molecular formula: C2H3BrO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HOBr


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Sample: CacothelineCAS No.: [561-20-6]Molecular formula: C21H21N3O7Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN

H

N

HH

HO

H

N+

OO

O

-O

O

HO


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Sample: Camostat　CAS No.: [59721-28-7]Molecular formula: C20H22N4O5Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

OHN

NHH2N

OO

N

O

H3CCH3


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Sample: L‐CarnitineCAS No.: [541-15-1]Molecular formula: C7H15NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Ammonium acetate =70/30 Flow rate: 1.0 ml/minTemperature: 30°CDetection: ELSDAttenuation: Gain=6,Atten=8 Sample conc.: 2.0mg/mlInjection volume: 1.5µlRetention time: 6.96minCapacity factor: 1.78

N+

H3C

HO H O

O-

H3CH3C


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Sample: CeftriaxoneCAS No.: [73384-59-5]Molecular formula: C18H18N8O7S3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH

H

N

S H

O

O

N

OHO

N

O

S

S

H3C

NH2N

N

N

CH3

OHO


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Sample: Chloroacetic AcidCAS No.: [79-11-8]Molecular formula: C2H3ClO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HOCl

7


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Sample: Citrazinic AcidCAS No.: [99-11-6]Molecular formula: C6H5NO4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Phosphate buffer(pH7)=50/50Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV220 nmAttenuation: 0.128 aufsSample conc.: 0.5mg/mlInjection volume: 0.5µlRetention time: 14.16minCapacity factor: 3.98

O OH

N OHHO


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Sample: CreatineCAS No.: [57-00-1]Molecular formula: C4H9N3O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


N

CH3

NH2

NH

O

HO


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Sample: CreatinineCAS No.: [60-27-5]Molecular formula: C4H7N3OColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


N

N

H2N

CH3

O


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Sample: Cyanoacetic AcidCAS No.: [372-09-8]Molecular formula: C3H3NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HO N


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Sample: Cyanuric AcidCAS No.: [108-80-5]Molecular formula: C3H3N3O3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH

HN

O

NH

O

O


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Sample: L‐CysteineCAS No.: [52-90-4]Molecular formula: C3H7NO2SColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2HHS OH

O

8


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Sample: L‐(-)‐CystineCAS No.: [56-89-3]Molecular formula: C6H12N2O4S2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2HS

SH2N H

O

HOOH

O


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Sample: CytidineCAS No.: [65-46-3]Molecular formula: C9H13N3O5Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HOH

HHHH

HO

N

N

NH2

O


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Sample: CytosineCAS No.: [71-30-7]Molecular formula: C4H5N3OColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


N

NH

O

NH2


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Sample: 3,4-Diaminobenzoic Acid CAS No.: [619-05-6]Molecular formula: C7H8N2O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O OH

H2NNH2


NACALAI TESQUE, INC

Sample: 3,5-Diaminobenzoic Acid CAS No.: [535-87-5]Molecular formula: C7H8N2O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O OH

NH2H2N


NACALAI TESQUE, INC

Sample: 2,4-DiaminophenolCAS No.: [95-86-3]Molecular formula: C6H8N2OColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 20mmol/l


NH2

HO

NH2

9


NACALAI TESQUE, INC

Sample: DL‐2,6‐Diaminopimelic AcidCAS No.: [583-93-7]Molecular formula: C7H14N2O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2NH2

O

OH

OH

O


NACALAI TESQUE, INC

Sample: DL‐2,3‐Diaminopropionic AcidCAS No.: [54897-59-5]Molecular formula: C3H8N2O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2

H2N OH

O


NACALAI TESQUE, INC

Sample: Diatrizoic AcidCAS No.: [117-96-4]Molecular formula: C11H9I3N2O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


I I

O OH

HN NH

IO CH3 OH3C


NACALAI TESQUE, INC

Sample: Dipicolinic acidCAS No.: [499-83-2]Molecular formula: C7H5NO4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


N

O OH

O

OH


NACALAI TESQUE, INC

Sample: DithiouracilCAS No.: [2001-93-6]Molecular formula: C4H4N2S2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN

NH

S

S


NACALAI TESQUE, INC

Sample: L‐DOPACAS No.: [59-92-7]Molecular formula: C9H11NO4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2H

HO

HO

OH

O

10


NACALAI TESQUE, INC

Sample: DopamineCAS No.: [51-61-6]Molecular formula: C8H11NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HO

NH2

OH


NACALAI TESQUE, INC

Sample: meso‐Erythritol 〔meso‐Erythrite〕CAS No.: [149-32-6]Molecular formula: C4H10O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ H2O=90/10Flow rate: 1.0 ml/minTemperature: 30°CDetection: ELSDAttenuation: Gain=6,Atten=8 Sample conc.: 1.0mg/mlInjection volume: 1.0µlRetention time: 5.78minCapacity factor: 1.18

OHH

HO H

HOOH


NACALAI TESQUE, INC

Sample: FamotidinCAS No.: [76824-35-6]Molecular formula: C8H15N7O2S3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


SNHO

O NH2 NH

S N

SNH

H2N

NH


NACALAI TESQUE, INC

Sample: Folic AcidCAS No.: [59-30-3]Molecular formula: C19H19N7O6Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 20mmol/l


HN H

O

O

OH

HN

NN

HNN

O

H2N

OH

O


NACALAI TESQUE, INC

Sample: Folinic AcidCAS No.: [58-05-9]Molecular formula: C20H23N7O7Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN H

O

O

OH

HN

NHNO

HNN

O

H2N

OH

O


NACALAI TESQUE, INC

Sample: FormamideCAS No.: [75-12-7]Molecular formula: CH3NOColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ H2O=95/5Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV220 nmAttenuation: 0.128 aufsSample conc.: 10.0mg/mlInjection volume: 0.5µlRetention time: 4.58minCapacity factor: 0.52

HCO

NH2

11


NACALAI TESQUE, INC

Sample: D‐Fructose‐6‐phosphateCAS No.: [643-13-0]Molecular formula: C6H13O9PColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Phosphate buffer(pH7)=50/50Flow rate: 1.0 ml/minTemperature: 30°CDetection: RIAttenuation: 4×10-5RIU/FSSample conc.: 10.0mg/mlInjection volume: 5.0µlRetention time: 12.16minCapacity factor: 3.64

OH

OHO

O

P OOH

HO

OH(CH2OH)


NACALAI TESQUE, INC

Sample: Fuchsine, AcidCAS No.: [3244-88-0]Molecular formula: C20H17N3Na2O9S3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


S

O

OO

Na

H2N

H3C

S

S

O

O

ONH2+

-O

O

O

NH2

Na


NACALAI TESQUE, INC

Sample: Fumaric AcidCAS No.: [110-17-8]Molecular formula: C4H4O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HOH

H

O

OH


NACALAI TESQUE, INC

Sample: Gluconic AcidCAS No.: [526-95-4]Molecular formula: C6H12O7Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


OHH

HO HHO H

OHH

O

OH

HO


NACALAI TESQUE, INC

Sample: α‐D‐Glucose‐1‐phosphateCAS No.: [59-56-3]Molecular formula: C6H13O9PColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

OH

OH

OHO

OH

P

OH

OOH


NACALAI TESQUE, INC

Sample: D‐Glucose‐6‐phosphateCAS No.: [56-73-5]Molecular formula: C6H13O9PColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

OH

OH

OH

OP O

OH

HO

H(OH)

12


NACALAI TESQUE, INC

Sample: D-Glucuronic AcidCAS No.: [6556-12-3]Molecular formula: C6H10O7Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

OH

OH

OH

OH

HO O


NACALAI TESQUE, INC

Sample: L‐Glutamic AcidCAS No.: [56-86-0]Molecular formula: C5H9NO4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2HHO

O

OH

O


NACALAI TESQUE, INC

Sample: L‐GlutamineCAS No.: [56-85-9]Molecular formula: C5H10N2O3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2H

O

H2N

O

OH


NACALAI TESQUE, INC

Sample: Glutaric AcidCAS No.: [110-94-1]Molecular formula: C5H8O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HO O

OH


NACALAI TESQUE, INC

Sample: DL‐Glyceric AcidCAS No.: [600-19-1]Molecular formula: C3H6O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


OHHHO OH

O


NACALAI TESQUE, INC

Sample: GlycinamideCAS No.: [598-41-4]Molecular formula: C2H6N2OColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

NH2

NH2

13


NACALAI TESQUE, INC

Sample: GlycineCAS No.: [56-40-6]Molecular formula: C3H7NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HONH2


NACALAI TESQUE, INC

Sample: Glycolic AcidCAS No.: [79-14-1]Molecular formula: C2H4O3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HOOH


NACALAI TESQUE, INC

Sample: GlycylglycineCAS No.: [556-50-3]Molecular formula: C4H8N2O3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN

O

H2N

O

OH


NACALAI TESQUE, INC

Sample: Guanidoacetic AcidCAS No.: [352-97-6]Molecular formula: C3H7N3O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN

NH

H2NO

OH


NACALAI TESQUE, INC

Sample: 1,2,6-HexanetriolCAS No.: [106-69-4]Molecular formula: C6H14O3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ H2O=90/10Flow rate: 1.0 ml/minTemperature: 30°CDetection: ELSDAttenuation: Gain=6,Atten=8 Sample conc.: 1.0mg/mlInjection volume: 2.0µlRetention time: 5.19minCapacity factor: 0.80

OH

OHHO


NACALAI TESQUE, INC

Sample: L-HistidineCAS No.: [71-00-1]Molecular formula: C6H9N3O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2HNHN OH

O

14


NACALAI TESQUE, INC

Sample: L‐HomocystineCAS No.: [626-72-2]Molecular formula: C8H16N2O4S2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2H

SS

H2N H

O

HOOH

O


NACALAI TESQUE, INC

Sample: L‐HomoserineCAS No.: [672-15-1]Molecular formula: C4H9NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2H

HOOH

O


NACALAI TESQUE, INC

Sample: Hydantoic AcidCAS No.: [462-60-2]Molecular formula: C3H6N2O3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HNO

NH2

O

OH


NACALAI TESQUE, INC

Sample: HydantoinCAS No.: [461-72-3]Molecular formula: C3H4N2O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HNNH

O

O


NACALAI TESQUE, INC

Sample: Hydroxylamine‐O‐sulfonic AcidCAS No.: [2950-43-8]Molecular formula: H3NO4SColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


SOHO

OONH2


NACALAI TESQUE, INC

Sample: cis-4-Hydroxy-D-proline CAS No.: [2584-71-6]Molecular formula: C5H9NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH

H O

HO

HOH

15


NACALAI TESQUE, INC

Sample: L‐HydroxyprolineCAS No.: [51-35-4]Molecular formula: C5H9NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH

H O

HO

HOH


NACALAI TESQUE, INC

Sample: N‐HydroxysuccinimideCAS No.: [6066-82-6]Molecular formula: C4H5NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NOH

OO


NACALAI TESQUE, INC

Sample: Indigo carmineCAS No.: [860-22-0]Molecular formula: C16H8N2Na2O8S2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


SO

O

ONa

O

NH

HN

OS

O

O

ONa


NACALAI TESQUE, INC

Sample: D-Isoascorbic AcidCAS No.: [89-65-6]Molecular formula: C6H8O6Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HHO

HHO

HO

HO O


NACALAI TESQUE, INC

Sample: Isocinchomeronic AcidCAS No.: [100-26-5]Molecular formula: C7H5NO4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


N

O OH

OHO


NACALAI TESQUE, INC

Sample: L-IsoleucineCAS No.: [73-32-5]Molecular formula: C6H13NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2H

HH3C

O

OH

16


NACALAI TESQUE, INC

Sample: Isonicotinic AcidCAS No.: [55-22-1]Molecular formula: C6H5NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O OH

N


NACALAI TESQUE, INC

Sample: IsonicotinohydrazideCAS No.: [54-85-3]Molecular formula: C6H7N3OColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NHO

NH2

N


NACALAI TESQUE, INC

Sample: Isopropyl β-D-1-thiogalactopyranosideCAS No.: [367-93-1]Molecular formula: C9H18O5SColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ H2O=90/10Flow rate: 1.0 ml/minTemperature: 30°CDetection: ELSDAttenuation: Gain=6,Atten=8 Sample conc.: 0.1mg/mlInjection volume: 0.5µlRetention time: 6.23minCapacity factor: 1.15

OH O S

OH

OH

OH CH3

CH3


NACALAI TESQUE, INC

Sample: Kojic AcidCAS No.: [501-30-4]Molecular formula: C6H6O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

OH

OHO


NACALAI TESQUE, INC

Sample: L-LeucineCAS No.: [61-90-5]Molecular formula: C6H13NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2H

H3C

CH3

O

OH


NACALAI TESQUE, INC

Sample: D-Leucyl-L-tyrosineCAS No.: [3303-29-5]Molecular formula: C15H22N2O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN H

O

NH2

H

CH3H3C

HO

OH

O

17


NACALAI TESQUE, INC

Sample: L‐LysineCAS No.: [56-87-1]Molecular formula: C6H14N2O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2H

H2NOH

O


NACALAI TESQUE, INC

Sample: Maleic AcidCAS No.: [110-16-7]Molecular formula: C4H4O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HOH

H

O OH


NACALAI TESQUE, INC

Sample: L‐(-)‐Malic AcidCAS No.: [97-67-6]Molecular formula: C4H6O5Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


OHHO

HOOH

O


NACALAI TESQUE, INC

Sample: Malonic AcidCAS No.: [141-82-2]Molecular formula: C3H4O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HO O

OH


NACALAI TESQUE, INC

Sample: Mecobalamin CAS No.: [13422-55-4]Molecular formula: C63H91CoN13O14PColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


N N

N N

H2N O

H2N

O

H2N

O

NH

O

O

N

NO OH

O

NH2

ONH2

O

NH2O

Co+

O

-O

HO

P


NACALAI TESQUE, INC

Sample: Metanilic AcidCAS No.: [121-47-1]Molecular formula: C6H7NO3SColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


SHO OO

H2N

18


NACALAI TESQUE, INC

Sample: L‐MethionineCAS No.: [63-68-3]Molecular formula: C5H11NO2SColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2H

SH3C OH

O


NACALAI TESQUE, INC

Sample: 6-Methyl-2-thiouracilCAS No.: [56-04-2]Molecular formula: C5H6N2OSColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN

NH

S

O

CH3


NACALAI TESQUE, INC

Sample: N‐MethylglucamineCAS No.: [6284-40-8]Molecular formula: C7H17NO5Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Citrate buffer(pH7.0)=70/30Flow rate: 1.0 ml/minTemperature: 30°CDetection: RIAttenuation: 4×10-5RIU/FS Sample conc.: 10.0mg/mlInjection volume: 2.0µlRetention time: 15.52minCapacity factor: 4.22

OHH

HO HHO H

OHH

HN

HO CH3


NACALAI TESQUE, INC

Sample: N‐MethylhydroxylamineCAS No.: [593-77-1]Molecular formula: CH5NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH

OHH3C


NACALAI TESQUE, INC

Sample: Mucic AcidCAS No.: [526-99-8]Molecular formula: C6H10O8Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


OHH

HO HHO H

OHHO

HO

O

OH


NACALAI TESQUE, INC

Sample: MurexideCAS No.: [3051-09-0]Molecular formula: C8H8N6O6Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


N

HN

O

NH

O-

O

NH

O

HN

O

O

NH4+

19


NACALAI TESQUE, INC

Sample: NicotinamideCAS No.: [98-92-0]Molecular formula: C6H6N2OColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


OH2N

N


NACALAI TESQUE, INC

Sample: Nicotinic AcidCAS No.: [59-67-6]Molecular formula: C6H5NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O OH

N


NACALAI TESQUE, INC

Sample: L‐NoradrenalineCAS No.: [51-41-2]Molecular formula: C8H11NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Citrate buffer(pH7.0)=60/40 Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV254 nmAttenuation: 0.128 aufsSample conc.: 5.0mg/mlInjection volume: 1.0µlRetention time: 5.47minCapacity factor: 1.07

OHH

NH2

HO

OH


NACALAI TESQUE, INC

Sample: DL-NorleucineCAS No.: [616-06-8]Molecular formula: C6H13NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


H3C OH

O

NH2


NACALAI TESQUE, INC

Sample: DL‐NorvalineCAS No.: [760-78-1]Molecular formula: C5H11NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


H3COH

O

NH2


NACALAI TESQUE, INC

Sample: L‐OrnithineCAS No.: [70-26-8]Molecular formula: C5H12N2O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2HH2N OH

O

20


NACALAI TESQUE, INC

Sample: Orotic AcidCAS No.: [65-86-1]Molecular formula: C5H4N2O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN

O OH

NH

O O


NACALAI TESQUE, INC

Sample: Oxalic AcidCAS No.: [144-62-7]Molecular formula: C2H2O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HOO

OH


NACALAI TESQUE, INC

Sample: Oxamic AcidCAS No.: [471-47-6]Molecular formula: C2H3NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HOO

NH2


NACALAI TESQUE, INC

Sample: OxytocinCAS No.: [50-56-6]Molecular formula: C43H66N12O12S2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH

H

H

HN

O

OH

NHHHN

O

O

H

O NH2

NH2HHN O

S

O

NH2

HO

SH

N

O

H

HN

O

H NH

O

O

NH2


NACALAI TESQUE, INC

Sample: D‐Pantothenic AcidCAS No.: [79-83-4]Molecular formula: C9H17NO5Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Ammonium acetate =50/50 Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV220 nmAttenuation: 0.128 aufs Sample conc.: 10.0mg/mlInjection volume: 1.0µlRetention time: 13.21minCapacity factor: 3.60

CH3H3C

HO H

HO

HN

O

O

OH


NACALAI TESQUE, INC

Sample: L-(-)-PhenylalanineCAS No.: [63-91-2]Molecular formula: C9H11NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2HOH

O

21


NACALAI TESQUE, INC

Sample: p‐PhenylenediamineCAS No.: [106-50-3]Molecular formula: C6H8N2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2

NH2


NACALAI TESQUE, INC

Sample: L‐(+)‐α‐PhenylglycineCAS No.: [2935-35-5]Molecular formula: C8H9NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


H2N H

O OH


NACALAI TESQUE, INC

Sample: PhosphocreatineCAS No.: [67-07-2]Molecular formula: C4H10N3O5PColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


P

HN

OH

HO

ON

NH

CH3 O

OH


NACALAI TESQUE, INC

Sample: O‐Phospho‐L‐serineCAS No.: [407-41-0]Molecular formula: C3H8NO6PColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 20mmol/l


PO

O

HO

HO

NH2H

O

OH


NACALAI TESQUE, INC

Sample: Picolinic acidCAS No.: [98-98-6]Molecular formula: C6H5NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


N

O OH


NACALAI TESQUE, INC

Sample: Pivalic AcidCAS No.: [75-98-9]Molecular formula: C5H10O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


CH3

CH3H3CO

OH

22


NACALAI TESQUE, INC

Sample: ProcaterolCAS No.: [72332-33-3]Molecular formula: C16H22N2O3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 20mmol/l


HNCH3

CH3

CH3

HN

HO

OOH


NACALAI TESQUE, INC

Sample: L-ProlineCAS No.: [147-85-3]Molecular formula: C5H9NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH

H

O

OH


NACALAI TESQUE, INC

Sample: Propionic AcidCAS No.: [79-09-4]Molecular formula: C3H6O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HOCH3


NACALAI TESQUE, INC

Sample: Pyruvic AcidCAS No.: [127-17-3]Molecular formula: C3H4O3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 20mmol/l


O

HO

O

CH3


NACALAI TESQUE, INC

Sample: Ribose‐5‐phosphateCAS No.: [4300-28-1]Molecular formula: C5H11O8PColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 20mmol/l

Phosphate buffer(pH7)=50/50 Flow rate: 1.0 ml/minTemperature: 30°CDetection: RIAttenuation: 4×10-5RIU/FS Sample conc.: 10.0mg/mlInjection volume: 5.0µlRetention time: 8.02minCapacity factor: 2.06

OH OH

OOP O

OH

HO

H(OH)


NACALAI TESQUE, INC

Sample: D‐Saccharic AcidCAS No.: [87-73-0]Molecular formula: C6H10O8Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


OHH

HO HHO H

OHHO

HOO

OH

23


NACALAI TESQUE, INC

Sample: SarcosineCAS No.: [107-97-1]Molecular formula: C3H7NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HONH

CH3


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Sample: Sebacic AcidCAS No.: [111-20-6]Molecular formula: C10H18O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HOO

OH


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Sample: L-SerineCAS No.: [56-45-1]Molecular formula: C3H7NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2HHO

O

OH


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Sample: SinigrinCAS No.: [3952-98-5]Molecular formula: C10H16KNO9S2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


OHO

HOOH

OH

S

NOS O

O

OK

CH2


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Sample: Succinic AcidCAS No.: [110-15-6]Molecular formula: C4H6O4Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


O

HOO

OH


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Sample: SulbactamCAS No.: [68373-14-8]Molecular formula: C8H11NO5SColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


++SO-

O-

CH3CH3

N

H

H

OHO

O

24


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Sample: Sulfanilic acidCAS No.: [121-57-3]Molecular formula: C6H7NO3SColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


S OHO

O

NH2


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Sample: L‐(+)‐Tartaric AcidCAS No.: [87-69-4]Molecular formula: C4H6O6Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


OHH

HO H

O

HOOH

O


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Sample: TaurineCAS No.: [107-35-7]Molecular formula: C2H7NO3SColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Ammonium acetate =70/30Flow rate: 1.0 ml/minTemperature: 30°CDetection: ELSDAttenuation: Gain=6,Atten=8Sample conc.: 1.0mg/mlInjection volume: 1.0µlRetention time: 6.40minCapacity factor: 1.25

SHO

OO

NH2


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Sample: L-TheanineCAS No.: [3081-61-6]Molecular formula: C7H14N2O3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2HNH

O

H3C OH

O


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Sample: 2‐Thiobarbituric AcidCAS No.: [504-17-6]Molecular formula: C4H4N2O2SColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH

HN

S O

O


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Sample: 2‐ThiouracilCAS No.: [141-90-2]Molecular formula: C4H4N2OSColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN

NH

S

O

25


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Sample: L-ThreonineCAS No.: [72-19-5]Molecular formula: C4H9NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2H

HO

H3C

H O

OH


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Sample: TodralazineCAS No.: [14679-73-3]Molecular formula: C11H12N4O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


HN

N

NH

N

OO

CH3


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Sample: Trichloroacetic Acid CAS No.: [76-03-9]Molecular formula: C2HCl3O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


Cl

ClCl

O

OH


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Sample: Tris(hydroxymethyl)aminomethaneCAS No.: [77-86-1]Molecular formula: C4H11NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Ammonium acetate =80/20Flow rate: 1.0 ml/minTemperature: 30°CDetection: ELSDAttenuation: Gain=6,Atten=8 Sample conc.: 2.0mg/mlInjection volume: 1.0µlRetention time: 6.47minCapacity factor: 1.48

NH2

OHHO

OH


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Sample: L-TryptophanCAS No.: [73-22-3]Molecular formula: C11H12N2O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l

Citrate buffer(pH7.0) =70/30Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV254 nmAttenuation: 0.128 aufsSample conc.: 0.5mg/mlInjection volume: 0.5µlRetention time: 5.14minCapacity factor: 0.95

HN H2N HO

OH


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Sample: L-TyrosineCAS No.: [60-18-4]Molecular formula: C9H11NO3Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2HHO

OH

O

26


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Sample: UracilCAS No.: [66-22-8]Molecular formula: C4H4N2O2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


N

NHO

OH


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Sample: UreaCAS No.: [57-13-6]Molecular formula: CH4N2OColumn: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ H2O=90/10Flow rate: 1.0 ml/minTemperature: 30°CDetection: UV210 nmAttenuation: 0.128 aufsSample conc.: 10.0mg/mlInjection volume: 2.0µlRetention time: 6.12minCapacity factor: 1.15

O

NH2H2N


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Sample: UridineCAS No.: [58-96-8]Molecular formula: C9H12N2O6Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH

O

ON

O

OHOH

HHHH

HO


NACALAI TESQUE, INC

Sample: L-ValineCAS No.: [72-18-4]Molecular formula: C5H11NO2Column: HILICColumn size: 4.6mmI.D.-250mmMobile phase: Acetonitrile/ 10mmol/l


NH2HH3C

CH3

OH

O

Date post:	12-Aug-2020
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Selection guide of mobile phase · (1) The effect of organic solvent type and content - In general,...

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