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HPLC CHROMATOGRAMS HPLC Chromatograms by Column Phase Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .709 Environmental . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .710-718 Aldehydes, Explosives, Herbicides, Ketones, Pesticides, Polycyclic Aromatic Hydrocarbons
Foods & Flavors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .719-735 Dietary Supplements, Drug Residues, Flavor Compounds, Food Contaminants, Preservatives, Sugars, Vitamins
Pharmaceutical . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .736-743 Admixed Drugs, Analgesics, Antiarrhythmics, Antiasthmatics, Antibiotics, Anti-Diabetic, Corticosteroids, Famotidine, Lactulose Concentrate, Potential Genotoxic Impurities, Steroids, Xanthines
Biochemical/Clinical . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .744-746 Amino Acids, Catecholamines, Nucleic Acid Bases, Nucleosides, Nucleotides, Oxytocin, Peptides, Proteins, Purines/Pyrimidines
Clinical/Forensic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .747-751 Alcohol Metabolites, Amphetamines, Benzodiazepines, Cocaine, Diuretics, Opiates, Pain Management Drugs, THC
Website : www.chromtech.net.au E-mail :
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HPLC CHROMATOGRAMS
HPLC Chromatograms by Column Phase HPLC Chromatograms by Column Phase (Alphabetical Order) Allure® AK
Ultra C8
carbonyls . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .181, 710
explosives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .711 oxycodone . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .738 vanillin & ethyl vanillin . . . . . . . . . . . . . . . . . . . . . . .728 vitamins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .733
Allure® Basix herbicides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .714
Allure® Biphenyl antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .740 corticosteroids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .741 steroids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .742
Ultra Carbamate
Allure® C18
antiarrhythmics . . . . . . . . . . . . . . . . . . . . . . . . . . . . .739
herbicides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .712 pesticides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .716
Ultra IBD
Allure® Organic Acids fruit juice acids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .531
Allure® PFP Propyl antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .723 benzodiazepines . . . . . . . . . . . . . . . . . . . . . . . . . . . . .751 catecholamines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .745 cocaine, ecgonine methyl ester . . . . . . . . . . . . . . . . .750 nucleic acid bases . . . . . . . . . . . . . . . . . . . . . . . . . . . .744 opiates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .748
food contaminants (carbamates) . . . . . . . . . . . . . . .719
Ultra Cyano
antiarrhythmics . . . . . . . . . . . . . . . . . . . . . . . . . . . . .739 antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .740, 741 diflubenzuron (pesticide) . . . . . . . . . . . . . . . . . . . . .716 glyburide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .741 nucleosides, nucleotides, & nucleic acid bases . . . .744 vitamins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .732
Ultra PFP purines, pyrimidines . . . . . . . . . . . . . . . . . . . . . . . . .745
Ultra Phenyl guaifenesin, codeine . . . . . . . . . . . . . . . . . . . . . . . . . .743
Pinnacle DB® Aqueous C18
Ultra Quat
food contaminants . . . . . . . . . . . . . . . . . . . . . . .720, 722 pesticides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .720, 722
paraquat, diquat . . . . . . . . . . . . . . . . . . . . . . . . .189, 715
Pinnacle® DB Biphenyl
dietary supplements . . . . . . . . . . . . . . . . . . . . . . . . . .729 herbicides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .713 melatonin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .729 organic acids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .729 vitamins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .734
drug residues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .725 steroids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .742
Pinnacle® DB C18 vanilla bean extracts . . . . . . . . . . . . . . . . . . . . . . . . . .727 xanthines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .743
Pinnacle® DB PAH PAHs
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .718
Pinnacle® II Amino lactulose concentrate . . . . . . . . . . . . . . . . . . . . . . . . .743 sugars . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .727
Pinnacle® II C18 allicin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .730 capsaicinoids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .728 morphine sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . .738 phenolic antioxidants . . . . . . . . . . . . . . . . . . . .727, 730
Pinnacle® II Cyano piperine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .728
Pinnacle® II PAH PAHs
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .717
Pinnacle® II Silica
Ultra II® Aqueous C18
GC & HPLC Chromatograms
Compound Index starting on page 752
Ultra II® Aromax explosives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .485, 711 famotidine and USP impurities . . . . . . . . . . . . . . . .736 potential genotoxic impurities . . . . . . . . . . . . . . . . .737 vitamins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .735
Ultra II® Biphenyl alcohol metabolites . . . . . . . . . . . . . . . . . . . . . . . . . .749 amphetamines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .750 antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .725 diuretics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .749 drug residues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .725 drugs of abuse . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .748 NSAIDs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .723 pain managment drugs in urine . . . . . . . . . . . . . . . .747 sulfa drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .725 THC & metabolites . . . . . . . . . . . . . . . . . . . . . . . . . .748
hydrocodone bitartrate . . . . . . . . . . . . . . . . . . . . . . .738 tocopherols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .730
Ultra II® C18
Ultra Aqueous C18
Ultra II® Carbamate
amino acids (aromatics) . . . . . . . . . . . . . . . . . . . . . .745 analgesics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .737 carboxylic acids . . . . . . . . . . . . . . . . . . . . . . . . . . . . .726 food contaminants . . . . . . . . . . . . . . . . . . . . . . .721, 722 herbicides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .714 pesticides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .721 phenethyl glucosinolate . . . . . . . . . . . . . . . . . . . . . . .730 sudan dyes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .722 vitamins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .731
food contaminants . . . . . . . . . . . . . . . . . . . . . . . . . . .719 pesticides (carbamates) . . . . . . . . . . . . . . . . . . . . . . .719
vitamins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .733
Viva C18 oxytocin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .746 peptides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .745 proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .746
Chromatogram Search Tool Search by compound name, synonym, CAS # or keyword
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Ultra C18 acetaminophen, narcotic analgesics . . . . . . . . . . . . .738 aldehydes, ketones . . . . . . . . . . . . . . . . . . . . . . . . . . .710 beclomethasone . . . . . . . . . . . . . . . . . . . . . . . . . . . . .739 corticosteroids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .741 drug residues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .724 herbicides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .712 hydrocodone bitartrate, acetaminophen . . . . . . . . .743 nitrofuran metabolites . . . . . . . . . . . . . . . . . . . . . . . .724 vitamins (fat soluble) . . . . . . . . . . . . . . . . . . . . . . . . .732
800-356-1688 or 814-353-1300
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HPLC CHROMATOGRAMS | ENVIRONMENTAL
Aldehydes, Ketones Carbonyls by CARB Method 1004 on Allure® AK
Sample: Inj.: Conc.:
10µL 3µg/mL each analyte, as aldehyde/ketone Sample diluent: acetonitrile Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase:
Flow: Temp.: Det.:
Ret. Time (min.)
Peak DNPH derivatives of: 1. formaldehyde 2. acetaldehyde 3. acrolein 4. acetone 5. propionaldehyde 6. crotonaldehyde 7. methacrolein 8. butyraldehyde 9. methylethylketone 10. benzaldehyde 11. valeraldehyde 12. m-tolualdehyde 13. hexaldehyde
Allure® AK 9159525-700 200mm x 4.6mm 5µm 60Å A) water : B) acetonitrile Time (min.) %B 0 60 8 70 10 100 1.5mL/min. 30°C UV @ 360nm
4.74 5.78 6.86 7.09 7.31 8.19 8.55 8.79 9.06 10.03 10.39 11.08 11.36
LC_EV0393
Aldehydes and Ketones (DNPH derivatives) on Ultra C18 (40°C) 1
Peak List: DNPH derivatives of: 1. formaldehyde 2. acetaldehyde 3. acetone 4. acrolein 5. propionaldehyde 6. crotonaldehyde 7. MEK 8. methacrolein 9. butyraldehyde 10. benzaldehyde 11. valeraldehyde 12. m-tolualdehyde 13. hexanaldehyde
Sample: Inj.: Conc.:
10µL 3.0µg/mL each derivative
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra C18 9174565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase:
2
Flow rate: Det.: Temp.:
4
Solvent A: water Solvent B: acetonitrile Solvent C: tetrahydrofuran 50-30% A/30-65% B/20-5% C, 0-15 min. 1.5mL/min. UV @ 365nm 40°C
3 5 8
6 7
9 10
11 13
12
0
2
4
6
8
10
12
14 min.
LC_0036
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HPLC CHROMATOGRAMS | ENVIRONMENTAL
Explosives Explosives EPA Method 8330B Ultra II® Aromax
1. 2. 3. 4. 5. 6. 7. 8. 9.
HMX RDX nitroglycerin nitrobenzene 4-amino-2,6-dinitrotoluene 3,5-dinitroaniline 2-nitrotoluene 2-amino-4,6-dinitrotoluene 3-nitrotoluene
10. 11. 12. 13. 14. 15. 16. 17.
PETN 4-nitrotoluene 1,3-dinitrobenzene 2,6-dinitrotoluene tetryl 2,4-dinitrotoluene 1,3,5-trinitrobenzene 2,4,6-trinitrotoluene
NEW!
Sample: Nitroaromatics and Nitramine Explosives by HPLC, EPA 8330B (cat.# 33204) Inj.: 10µL Conc.: 10µg/mL each component Sample diluent: methanol Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra II® Aromax 9607575 250mm x 4.6mm 5µm 100Å
Conditions: Instrument: Mobile phase: Flow: Temp.: Det.:
Shimadzu Prominence UFLCXR water:methanol, 35:65 (v/v) 1.2mL/min. 30°C UV @ 254nm and 210nm
LC_EV0484
Explosives EPA Method 8330B Ultra C8
1. 2. 3. 4. 5. 6. 7. 8. 9.
HMX RDX nitroglycerin nitrobenzene 4-amino-2,6-dinitrotoluene 3,5-dinitroaniline 2-nitrotoluene 2-amino-4,6-dinitrotoluene 3-nitrotoluene
10. 11. 12. 13. 14. 15. 16. 17.
PETN 4-nitrotoluene 1,3-dinitrobenzene 2,6-dinitrotoluene tetryl 2,4-dinitrotoluene 1,3,5-trinitrobenzene 2,4,6-trinitrotoluene
NEW!
Sample:
Nitroaromatics and Nitramine Explosives by HPLC, EPA 8330B (cat.# 33204) Inj.: 10µL Conc.: 50µg/mL each component Sample diluent: methanol Column: Dimensions: Particle size: Pore size:
Ultra C8 cat.# 9103575 250mm x 4.6mm 5µm 100Å
Conditions: Instrument: Mobile phase: Flow: Temp.: Det.:
Shimadzu Prominence water:methanol, 52:48 (v/v) 1.2mL/min. 30°C UV @ 254nm and 210nm
LC_EV0485
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HPLC CHROMATOGRAMS | ENVIRONMENTAL
Herbicides Herbicides (Phenylurea) on Ultra C18 Peak List: 1. tebuthiuron 2. thidiazuron 3. monuron 4. fluometuron 5. diuron 6. propanil 7. siduron 8. linuron 9. carbazole 10. diflubenzuron
Conc. (µg/mL or ppm) 2 2 5 2 2 2 2 2 5 2
Sample: Inj.: Conc.: Sample diluent:
20µL see peak list acetonitrile
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra C18 9174565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase:
Flow: Temp.: Det.:
A: 25mM phosphate buffer, pH 2.5 B: acetonitrile Time (min.) %B 0 40 8 40 14 55 18 55 19 40 1.5mL/min. ambient UV @ 245nm
LC_0319
Herbicides on Allure® C18
Peak List: 1. cyanazine 2. methabenzthiazuron 3. chlortoluron 4. secbumeton 5. isoproturon 6. diuron 7. linuron 8. flusilazole 9. ethofumesate 10. neburon 11. pendimethalin
Ret. Time (min.) 12.04 13.88 14.59 15.74 16.18 16.78 22.48 24.29 24.53 25.74 31.46
Sample: Inj.: Conc.: Diluent:
5µL custom mix approx. 20µg/mL each acetonitrile
Column: Cat.#: Dimensions: Particle size: Pore size:
Allure® C18 9164565 150mm x 4.6mm 5µm 60Å
Conditions: Mobile phase:
Flow: Temp.: Det.:
A: water B: acetonitrile Time (min.) %B 0 20 10 40 15 40 26 80 34 80 34.1 20 1.2mL/min. ambient UV @ 220nm
LC_0309
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HPLC CHROMATOGRAMS | ENVIRONMENTAL
Herbicides Phenoxyacid Herbicides on Ultra II® Aqueous C18 (LC/MS/MS)
NEW! Peaks RT (min.) 1. Dicamba 1.14 2. Bentazon 1.32 3. 2,4-D 3.10 4. 3,5-DCBA 3.10 5. 4-Nitrophenol 3.10 6. MCPA 3.16 7. Dichlorprop 4.02 8. 2,4,5-T 4.46 9. Dinoseb 5.03 10. 2,4-DB 5.18 11. 2,4,5-TP 5.20 12. Acifluorfen 5.67 13. Pentachlorophenol 6.20
MRM 1 219.0 → 175.0 239.0 → 132.0 218.9 → 161.0 189.0 → 144.8 138.1 → 108.1 199.0 → 141.0 233.0 → 161.0 252.9 → 194.9 239.0 → 133.9 247.0 → 161.0 266.9 → 195.0 359.9 → 316.1 264.8 → 35.0
MRM 2 221.0 → 177.0 239.0 → 175.0 220.9 → 163.0 189.0 → 59.0 138.1 → 46.0 201.0 → 143.0 235.0 → 163.0 254.9 → 196.9 239.0 → 193.8 249.0 → 163.0 266.9 → 159.0 359.9 → 195.0 264.8 → 37.0
Column Dimensions: Particle Size: Pore Size: Temp.:
Ultra II® Aqueous C18 (cat.# 9608312) 100 mm x 2.1 mm ID 3 µm 100 Å 35 °C
Sample Diluent: Conc.: Inj. Vol.:
phenoxyacid herbicides acetonitrile 100 ppb 10 µL
Mobile Phase A: B:
10mM ammonium acetate in water 10mM ammonium acetate in methanol Time (min.) 0 8 12 14.8 15
LC_EV0511
%A 80 10 0 0 80
%B 20 90 100 100 20
Flow: 0.5 mL/min. Detector Applied Biosystems/MDS Sciex LC/MS/MS Model #: 4000 QTRAP™ LC/MS/MS System Ion Source: Electrospray Ion Mode: ESIIon Spray Voltage: -4.2 kV ESI Voltage: -4.2 V Gas 1: 50 psi (344.7 kPa) Gas 2: 60 psi (413.7 kPa) Source Temp.: 600 °C Mode: MRM Instrument Applied Biosystems/MDS Sciex LC/MS/MS System Acknowledgement Work performed at Applied Biosystems
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HPLC CHROMATOGRAMS | ENVIRONMENTAL
Herbicides Phenoxyacid Herbicides on Ultra Aqueous C18
Mix A and Mix B 1. picloram 2. chloramben 3. 4-nitrophenol 4. dicamba 5. bentazon 2,4-dichlorophenyl acetic acid (SS) (not included) 7. 2,4-D 8. MCPA 9. 3,5-dichlorobenzoic acid 10. MCPP 11. dichlorprop 12. 2,4,5-T 13. 2,4-DB 14. 2,4,5-TP (Silvex) 15. acifluorfen 1,4-dichlorobenzene (IS) (not included) 17. dinoseb 18. pentachlorophenol
Sample: Sample:
Chlorinated Acids by HPLC, Mix A (cat.# 32431), or Chlorinated Acids by HPLC, Mix B (cat.# 32430) 1,000µg/mL each component in acetonitrile Inj.: 10µL Conc.: 10ppm each analyte Sample diluent: acetonitrile Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase:
Flow: Temp.: Det.:
Ultra Aqueous C18 9178565 150mm x 4.6mm 5µm 100Å A) 0.05% H3PO4 B) acetonitrile Time (min.) %B 0 20.0 28 80.0 33 90.0 34 20.0 1.0mL/min. ambient UV @ 225nm
LC_EV0356
LC_EV0355
Phenoxyacid Herbicides on Allure® Basix Mix A and Mix B 1. picloram 2. chloramben 3. 4-nitrophenol 4. dicamba 5. bentazon 2,4-dichlorophenyl acetic acid (SS) (not included) 7. 2,4-D 8. MCPA 9. 3,5-dichlorobenzoic acid 10. MCPP 11. dichlorprop 12. 2,4,5-T 13. 2,4-DB 14. 2,4,5-TP (Silvex) 15. acifluorfen 1,4-dichlorobenzene (IS) (not included) 17. dinoseb 18. pentachlorophenol
Sample: Sample:
Chlorinated Acids by HPLC, Mix A (cat.# 32431), or Chlorinated Acids by HPLC, Mix B (cat.# 32430) 1,000µg/mL each component in acetonitrile Inj.: 10µL 10ppm each analyte Conc.: Sample diluent: acetonitrile
Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase:
Flow: Temp.: Det.:
Allure® Basix 9161565 150mm x 4.6mm 5µm 60Å A) 0.05% H3PO4 B) acetonitrile Time (min.) %B 0 20.0 28 80.0 33 90.0 34 20.0 1.0mL/min. ambient UV @ 225nm
LC_EV0357 LC_EV0358
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HPLC CHROMATOGRAMS | ENVIRONMENTAL
Herbicides Paraquat and Diquat on Ultra Quat
2 1
Peak List: 1. diquat 2. paraquat
Peak time (min.) 7.5 8.8
min.
Sample: Inj.: Conc.:
diquat 308nm 100µg/mL
20µL 20ppm each component (above); 20µg/mL, 40µg/mL, and 100µg/mL each component at 308nm (left) and at 257nm (right) Sample diluent: water Column: Cat.#: Dimensions: Particle size: Conditions: Mobile phase:
40µg/mL
Flow: Temp.: Det.:
paraquat 257nm 100µg/mL
Ultra Quat 9181565 150mm x 4.6mm 5µm Ultra Quat Reagent Solution (cat.# 32441): acetonitrile, 95:5 (v/v) 1.0 mL/min. 27°C UV @ 257nm (paraquat) UV @ 308nm (diquat)
40µg/mL
20µg/mL
20µg/mL
LC_0321
min.
Paraquat and Diquat on Ultra Quat
Fast, sensitive LC/MS/MS analysis!
Peak List 1. diquat 2. paraquat
paraquat dichloride [CAS# 1910-42-5]
diquat dibromide [CAS # 85-00-7]
LC_EV0383
*Data courtesy of Houssain El Aribi, Ph.D., LC/MS Product and Application Specialist, MDS SCIEX, 71 Four Valley Drive, Concord, Ontario, Canada, L4K 4V8
Sample: Inj.: Conc.: Sample diluent: Sample temp.:
10µL 5µg/mL each component DI water ambient
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra Quat 5181352 50mm x 2.1mm 3µm 100Å
Conditions: Mobile phase: Flow: Temp.: Det.: Interface: Ion Mode: Temp.: Ion Spray voltage: Collision exit potential: Curtain Gas: Gas supply 1: Gas supply 2: Quantitation: Q1/Q3: Dwell time:
10mM heptafluorobutyric acid:acetonitrile, 95:5 (v/v) 0.3mL/min. ambient Applied Biosystems/MDS SCIEX API 3200™ MS/MS system electrospray positive 600°C 5,500V 3V 15psi (103kPa) 70psi (483kPa) 60psi (414kPa) (MRM) unit resolution 200 ms
Precursor Ion Fragment Ion (amu) (amu) diquat, 183+ 157+ paraquat, 93 (2+) 171+
DP (V) 30 20
Collision Energy (eV) 30 20
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HPLC CHROMATOGRAMS | ENVIRONMENTAL
Pesticides Pesticides on Allure® C18 Peak List: 1. desisopropyl atrazine 2. desethyl atrazine 3. simazine 4. terbuthylazine desethyl 5. cyanazine 6. methabenzthiazuron 7. chlortoluron 8. atrazine 9. secbumeton 10. isoproturon 11. diuron 12. terbumeton 13. terbuthylazine 14. linuron 15. flusilazole 16. ethofumesate 17. neburon
Sample: Inj.: Conc.: Solvent:
5µL approx. 10ppm each pesticide acetonitrile
Column: Cat.#: Dimensions: Particle size: Pore size:
Allure® C18 9164565 150mm x 4.6mm 5µm 60Å
Conditions: Mobile phase
Flow: Temp.: Det.:
A: 20mM K2HPO4, pH 7.0 B: acetonitrile Time (min.) %B 0 20 10 40 15 40 30 70 30.1 20 1.2mL/min. ambient UV @ 220nm
LC_0310
Diflubenzuron (Pesticide) on Ultra IBD 1
Peak List: 1. diflubenzuron 2. N,N'-bis-(chlorobenzyl)urea Sample: Inj.: Conc.: Solvent:
10µL 1mg/mL water:acetonitrile (1:1, v/v)
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra IBD 9175365 150mm x 4.6mm 3µm 100Å
Jamie Hubler, Customer Service
Conditions: Mobile phase:
water:acetonitrile (45:55, v/v) 65% 0.1M ammonium acetate pH 6.0 1.0mL/min. ambient UV @ 260nm
Solvent A: Flow rate: Temp.: Det.:
Restek Customer Service 2
In the U.S. Call: 800-356-1688 (ext. 3) or 814-353-1300 (ext. 3) Monday–Friday 8:00 a.m.–6:00 p.m. ET Fax: 814-353-1309—24-hours a day Online: www.restek.com—24-hours a day
Outside the U.S.
0
2
4
6
8
10
12
14 min.
Contact your Restek representative: Refer to our list on pages 4-5 or visit our website at www.restek.com
LC_0081
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HPLC CHROMATOGRAMS | ENVIRONMENTAL
Polycyclic Aromatic Hydrocarbons 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16.
Polycyclic Aromatic Hydrocarbons Pinnacle® II PAH
6
Baseline resolution of PAHs in methanol!
Sample: Inj.: 10µL Conc.: 10µg/mL each component Sample diluent: methanol
naphthalene acenaphthylene acenaphthene fluorene phenanthrene anthracene fluoranthene pyrene benzo(a)anthracene chrysene benzo(b)fluoranthene benzo(k)fluoranthene benzo(a)pyrene benzo(ghi)perylene dibenzo(a,h)anthracene indeno(1,2,3-cd)pyrene * contaminant
Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Instrument: Mobile phase:
Flow: Temp.: Det.:
NEW!
Pinnacle II PAH 9219463 150mm x 3.2mm 4µm 110Å ®
Shimadzu Prominence A: water B: methanol Time (min.) %B 0.0 75 10.0 90 22.0 100 1.0mL/min. 30°C UV @ 254nm
5
10
9
4
11 13 12
7
16
8
1
14 15
2 3
*
0
2
4
6
8
10
12
14
16
18
20
22
24
LC_EV0480
Polycyclic Aromatic Hydrocarbons on Pinnacle® II PAH EPA 8310 – 18 component standard + benzo(j)fluoranthene Peak List: Retention Time 1. naphthalene 6.19 2. acenaphthylene 7.16 3. 1-methylnaphthalene 7.78 4. 2-methylnaphthalene 8.17 5. acenaphthene 8.46 6. fluorene 8.82 7. phenanthrene 9.64 8. anthracene 10.39 9. fluoranthene 11.22 10. pyrene 11.75 11. benzo(a)anthracene 13.55 12. chrysene 13.91 13. benzo(j)fluoranthene 14.92 14. benzo(b)fluoranthene 15.27 15. benzo(k)fluoranthene 15.82 16. benzo(a)pyrene 16.35 17. dibenzo(a,h)anthracene 17.10 18. benzo(ghi)perylene 17.67 19. indeno(1,2,3-cd)pyrene 17.99
Sample: Inj.: 10µL Conc.: 20µg/mL each component Sample diluent: acetonitrile Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Instrument: Mobile phase:
Flow: Temp.: Det.:
Pinnacle® II PAH 9219463 150mm x 3.2mm 4µm 110Å Shimadzu Prominence HPLC A: purified water B: acetonitrile Time (min.) %B 0 40 7 60 16 100 18.9 100 19 40 1.2mL/min. 30°C UV @ 254nm
LC_EV0449
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HPLC CHROMATOGRAMS | ENVIRONMENTAL
Polycyclic Aromatic Hydrocarbons EPA Method 610 Polycyclic Aromatic Hydrocarbons on Pinnacle® DB PAH Peak List: 1. naphthalene 2. acenaphthylene 3. 1-methylnaphthalene 4. 2-methylnaphthalene 5. acenaphthene 6. fluorene 7. phenanthrene 8. anthracene 9. fluoranthene 10. pyrene 11. benzo(a)anthracene 12. chrysene 13. benzo(b)fluoranthene 14. benzo(k)fluoranthene 15. benzo(a)pyrene 16. dibenzo(a,h)anthracene 17. benzo(ghi)perylene 18. indeno(1,2,3-cd)pyrene
LC_EV0469
Conditions: Mobile phase:
Sample: Inj.: 2µL Conc.: 20µg/mL each component Sample diluent: acetonitrile Column: Cat.#: Dimensions: Particle size: Pore size:
Pinnacle® DB PAH 9470252 50mm x 2.1mm 1.9µm 140Å
Flow: Temp.: Det.:
A: water B: acetonitrile Time (min.) %B 0 50 1 60 3 100 5 100 0.6mL/min. 30°C UV @ 254nm
EU 256/2005 Polycyclic Aromatic Hydrocarbons on Pinnacle® DB PAH
Peak List: 1. cyclopenta(c,d)pyrene 2. benzo(a)anthracene 3. chrysene 4. 5-methylchrysene 5. benzo(j)fluoranthene 6. benzo(b)fluoranthene 7. benzo(k)fluoranthene 8. benzo(a)pyrene 9. dibenzo(a,l)pyrene 10. dibenzo(a,h)anthracene 11. benzo(ghi)perylene 12. indeno(1,2,3-cd)pyrene 13. dibenzo(a,e)pyrene 14. dibenzo(a,i)pyrene 15. dibenzo(a,h)pyrene
min. LC_EV0470
Sample: Inj.: 2µL Conc.: 20µg/mL each component Sample diluent: acetonitrile Column: Cat.#: Dimensions: Particle size: Pore size:
Conditions: Mobile phase:
Pinnacle® DB PAH 9470252 50mm x 2.1mm 1.9µm 140Å Flow: Temp.: Det.:
A: water B: acetonitrile Time (min.) %B 0 50 1 90 2 95 5 100 7 100 0.6mL/min. 30°C UV @ 254nm
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HPLC CHROMATOGRAMS | FOODS & FLAVORS
Food Contaminants Carbamate Pesticides on Ultra II® Carbamate (LC/MS/MS)
NEW! Peaks RT (min.) 1. Aldicarb sulfone 1.9 2. Aldicarb sulfoxide 2.2 3. Oxamyl 2.4 4. Methomyl 2.6 5. 3-Hydroxycarbofuran 3.5 6. Aldicarb 4.5 7. Propoxur 5.1 8. Carbofuran 5.3 9. Carbaryl 5.8 10. Methiocarb 7.0
Transition 1 223.1 → 86.1 207.1 → 132.1 237.1 → 72.1 163.1 → 88.1 238.1 → 163.1 208.1 → 116.0 210.1 → 111.0 222.1 → 165.2 202.1 → 145.1 226.1 → 169.2
Transition 2 223.1 → 148.0 207.1 → 89.1 237.1 → 90.1 163.1 → 106.0 238.1 → 181.2 208.1 → 88.9 210.1 → 168.1 222.1 → 123.0 202.1 → 127.1 226.1 → 121.0
Column Dimensions: Particle Size: Pore Size: Temp.:
Ultra II® Carbamate (cat.# 9611312) 100 mm x 2.1 mm ID 3 µm 100 Å 35 °C
Sample
531.1 Carbamate Pesticide Calibration Mixture (cat.# 32273) acetonitrile 100 ppb each pesticide 5 µL
Diluent: Conc.: Inj. Vol.:
8
Mobile Phase A: B:
Flow:
10 mM NH4OAc in water 10 mM NH4OAc in methanol Time (min.) %B 0 10 1 10 8 90 10 90 11 10 0.5 mL/min.
Instrument Detector Model #: Ion Source: Ion Mode: Ion Spray Voltage: Gas 1: Gas 2: Source Temp.:
Applied Biosystems/MDS Sciex LC/MS/MS System Applied Biosystems/MDS Sciex LC/MS/MS 4000 QTRAP™ LC/MS/MS system TurboIonSpray® ESI+ 5 kV 40 psi (275.8 kPa) 60 psi (413.7 kPa) 500 °C
9
3
7
4 10 2
6
5 1
0.0
1.0
2.0
3.0
4.0
5.0 Time (min.)
6.0
7.0
8.0
9.0
10.0
LC_EV0510
Carbamates in Orange Oil on Ultra Carbamate (LC-TOFMS) Sample:
Peak List: 1. aldicarb sulfone 2. aldicarb sulfoxide 3. oxamyl 4. methomyl 5. 3-hydroxycarbofuran 6. aldicarb 7. propoxur 8. carbofuran 9. carbaryl 10. methiocarb 11. BDMC (IS)
Inj.: Conc.: Sample diluent: Matrix:
531.1 Carbamate Pesticide Calibration Mix (cat.# 32273) and Internal Standard 4-bromo-3,5-dimethylphenyl-N-methylcarbamate (cat.# 32274) 50:50 spiked into unprocessed orange oil at 10ppm 3µL 10ppm methanol orange oil
Column: Cat. #: Dimensions: Particle size: Pore size:
Ultra Carbamate 9177352 50mm x 2.1mm 3µm 100Å
Conditions: Mobile phase:
Flow: Temp.: Det.: Interface: Ion mode: Temp.: Nebulizer pressure: Desolvation gas (N2): Interface temperature: Nozzle: Capillary:
A: 2mM ammonium acetate:methanol, 90:10 B: 2mM ammonium acetate:methanol, 10:90 Time (min.) %B 0 20 20 100 25 100 200µL/min. ambient LECO Unique® LC-TOFMS ESI Positive 130°C 100kPa 4,000cc/min. 120°C (+) 62V (+) 2.75kV
LC_FF0472
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HPLC CHROMATOGRAMS | FOODS & FLAVORS
Food Contaminants Pesticides on Pinnacle® DB Aqueous C18 (LC/MS/MS, ESI+) Sample:
multicomponent pesticide standard Inj.: 5µL Conc.: 33.3ppb each pesticide Sample diluent: acetonitrile
NEW!
Column: Cat.#: Dimensions: Particle size: Pore size:
Pinnacle® DB Aqueous C18 9418252 50mm x 2.1mm 1.9µm 140Å
LC_EV0479A
Conditions: Mobile phase:
A: 10mM NH4OAc in water B: 10mM NH4OAc in methanol Time (min.) 0.0 1.0 8.0 10.0 11.0
Flow: Temp.: Instrument: Det.:
%B 10 10 90 90 10
Peak List: Compound methamidophos acephate bentazone omethoate bromoxynil butoxycarboxin aldicarb sulfoxide methomyl Ioxynil thiamethoxam monocrotophos dimethoate fenuron dioxacarb dicrotophos vamidothion carbendazim aldicarb formetanate acetamiprid carbetamide oxamyl aminocarb propoxur oxadixyl bendiocarb thiacloprid thiophanate methyl methabenzhiazuron tricyclazole pyracarbolid propamocarb carboxin ethiofencarb carbaryl thiabendazole carbanilide clethodim tebuthiuron
Polarity positive positive negative positive negative positive positive positive negative positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive
600µL/min. 35°C Shimadzu Prominence® UFLCXR Applied Biosystems 4000 QTRAP® LC/MS/MS system Ion Source: TurboIonSpray®, ESI+ IonSpray Voltage, ESI voltage: 5kV (ESI+) Gas 1: 40psi Gas 2: 60psi Source Temp.: 500°C; max pressure ~7,200psi
Mol. Wt. 141.13 183.17 240.28 213.14 276.91 222.27 206.26 162.21 370.92 291.7 223.17 229.26 164.21 223.23 237.19 287.34 191.19 190.27 221.26 222.67 236.27 219.26 208.26 209.24 278.31 223.23 252.71 342.4 221.28 189.23 217.27 188.27 235.3 225.31 201.22 201.25 212.25 359.91 228.32
RT (min.) 0.45 0.62 0.77 1.07 1.32 1.64 2.03 2.08 2.46 2.7 2.73 2.9 3.1 3.12 3.28 3.45 3.73 3.73 3.74 3.95 3.99 3.99 4.11, 4.44 4.15 4.21 4.24 4.25 4.3 4.31 4.35 4.38 4.41 4.51 4.52 4.58 4.6 4.6 4.61 4.68
Transition 1 142.0 → 94.0 184.0 → 142.8 239.0 → 131.8 214.0 → 124.9 275.9 → 80.8 223.1 → 86.1 207.1 → 132.1 163.1 → 88.1 369.8 → 126.7 292.0 → 211.0 224.1 → 127.0 230.1 → 199.0 165.1 → 72.1 224.0 → 123.0 238.0 → 112.0 288.0 → 146.0 192.0 → 160.0 208.1 → 116.0 222.1 → 165.2 223.1 → 126.1 237.0 → 192.0 237.1 → 72.2 209.2 → 137.1 210.1 → 111.0 279.0 → 219.0 224.1 → 109.1 253.2 → 126.1 343.0 → 151.0 222.0 → 165.1 190.0 → 163.0 218.2 → 125.0 189.2 → 102.2 236.0 → 143.0 226.1 → 106.9 202.1 → 145.1 202.0 → 175.0 213.1 → 94.0 360.0 → 164.0 229.2 → 172.4
Compound fluometuron metobromuron flutriafol chlortoluron simetryn pirimicarb forchlorfenuron siduron ethofumesate cycluron diuron diethofencarb furalaxyl methiocarb secbumeton terbumeton prometon paclobutrazol promecarb dimoxystrobin ametryn methoprotryne triadimefon mexacarbate fludioxinil boscalid azoxystrobin methoxyfenozide cyproconazole prothioconazole flufenacet butafenacil myclobutanil dimethomorph bromuconazole 47 prometryn fipronil triticonazole tetraconazole uniconazole fluoxastrobin cyazofamid terbutryn tebufenozide chloroxuron picoxystrobin carfentrazone-ethyl diclobutrazol fenoxycarb diflubenzuron neburon flusiazole epoxiconazole fenbuconazole bupirimate zoxamide rotenone benalaxyl tebuconazole penconazole cyprodinil propiconazole alanycarb triflumuron thiobencarb pyraclostrobin benzoximate pinoxaden clofentezine trifloxystrobin prochloraz difenoconazole benfuracarb triflumizole buprofezin piperonyl butoxide pyriproxyfen propargite etoxazole amitraz noviflumuron pyridaben fenpyroximate fenpropimorph spiroxamine spinosyn A
Polarity positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive negative negative positive positive positive negative positive positive positive positive positive positive negative positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive negative positive positive positive positive positive
Mol. Wt. 232.21 259.1 301.3 212.68 213.31 238.29 247.68 232.32 286.35 198.31 233.1 267.33 301.34 225.31 225.29 225.29 225.29 293.8 207.27 326.39 227.33 271.39 293.75 222.28 248.19 343.21 403.3 368.47 291.78 344.26 363.34 474.82 288.78 387.86 377.06 241.36 437.15 317.81 372.12 291.78 458.83 324.78 241.36 352.47 290.75 367.25 412.17 328.21 301.34 310.67 275.18 315.4 329.76 336.82 316.41 336.54 394.42 325.41 307.82 284.19 225.29 342.22 399.52 358.68 257.78 387.83 363.8 400.52 303.15 408.38 376.67 406.28 410.53 345.75 305.44 338.45 321.28 350.48 359.42 293.41 529.1 364.94 421.5 303.49 297.48 731.97
RT (min.) 4.75 4.82 4.84 4.92 4.96 4.97 5.04 5.09 5.11 5.13 5.13 5.15 5.18 5.24 5.24 5.25 5.26 5.26 5.27 5.27 5.29 5.32 5.32 5.34 5.34 5.36 5.36 5.38 5.39 5.41 5.46 5.47 5.51 5.52, 5.54 5.54 5.55 5.56 5.58 5.6 5.62 5.62 5.63 5.65 5.65 5.67 5.67 5.69 5.71 5.72 5.72 5.74 5.76 5.76 5.76 5.77 5.77 5.79 5.83 5.84 5.93 5.96 5.96 5.98 5.99 6 6 6.01 6.01 6.11 6.12 6.21 6.29 6.29 6.38 6.4 6.42 6.52 6.68 6.87 6.89 7.08 7.18 7.2 7.38 7.63, 7.81 8.37
Transition 1 233.1 → 72.0 259.0 → 170.2 302.0 → 123.0 213.1 → 72.2 214.0 → 124.0 239.2 → 72.1 248.0 → 129.1 233.3 → 137.2 304.0 → 121.0 199.1 → 89.1 233.1 → 72.0 268.0 → 226.0 302.2 → 95.0 226.1 → 169.2 226.2 → 169.9 226.2 → 169.9 226.2 → 142.3 294.0 → 70.0 208.2 → 109.2 327.1 → 205.0 228.2 → 186.2 272.2 → 240.2 294.0 → 197.0 223.2 → 166.2 247.0 → 179.9 343.0 → 307 404.1 → 372.1 369.0 → 149.0 292.2 → 70.2 341.9 → 305.8 364.0 → 152.0 492.2 → 331.1 289.0 → 70.0 388.0 → 301.0 378.0 → 159.0 242.2 → 158.1 434.9 → 329.8 318.0 → 70.0 372.0 → 159.0 292.2 → 70.2 459.2 → 427.2 325.0 → 108.0 242.2 → 186.0 353.1 → 133.1 291.0 → 72.0 368.0 → 145.0 412.0 → 346.0 328.0 → 70.0 302.0 → 88.1 311.2 → 158.1 275.0 → 88.0 316.0 → 247.0 330.0 → 121.0 337.0 → 125.0 317.0 → 166.0 336.2 → 187.1 395.0 → 213.0 326.0 → 148.0 308.0 → 70.0 284.0 → 159.0 226.0 → 93.0 342.0 → 159.0 400.1 → 238.2 359.1 → 156.1 258.1 → 125.0 388.0 → 194.0 364.0 → 199.0 401.3 → 317.2 303.0 → 138.0 409.2 → 186.0 376.0 → 308.0 406.0 → 251.0 411.2 → 195.1 346.1 → 278.1 306.0 → 201.0 356.2 → 177.2 322.0 → 96.0 368.0 → 231.0 360.1 → 141.0 294.2 → 163.3 527.1 → 343.8 365.0 → 147.0 422.0 → 366.0 304.0 → 147.0 298.0 → 144.0 732.6 → 142.2
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HPLC CHROMATOGRAMS | FOODS & FLAVORS
Food Contaminants Pesticides on Ultra Aqueous C18 (LC/MS/MS)
NEW!
A: Pesticides in positive ion mode
Sample: Inj.: Conc.: Sample diluent:
multicomponent pesticide standard 10µL 1ppb each pesticide acetonitrile
Column: Cat. #: Dimensions: Particle size: Pore size:
Ultra Aqueous C18 9178312 100mm x 2.1mm 3µm 100Å
Conditions: Instrument: Mobile phase:
Flow: Temp.:
IonSpray Voltage: Gas 1: Gas 2: Source Temp.:
1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
9.0
10.0
9.0
10.0
11.0
12.0
13.0
14.0
15.0
Time (min.)
LC_FF0490
Shimadzu Prominence® UFLCXR A: 10 mM NH4OAc in water B: 10 mM NH4OAc in methanol Time (min.) 0.0 8.0 12.0 14.8 14.9
Det.: Ion Source:
0.0
%B 20 90 100 100 20
B: Pesticides in negative ion mode
500µL/min 35°C
Applied Biosystems 4000 QTRAP® LC/MS/MS system TurboIonSpray® A & C: ESI+ B: ESI5kV (ESI+), -4.2kV (ESI-) 50psi 60psi 600°C 0.0 1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
11.0
12.0
13.0
14.0
15.0
Time, min
propamocarb
LC_FF0490A
omethoate
C: Improved retention of difficult pesticides in positive ion mode
Visit www.restek.com/cat005 to view our "Comprehensive Pesticide Residue Analysis by LC/MS/MS Using an Ultra Aqueous C18 Column" article (contains compound identifications and MS details).
methamidophos acephate
for more info
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
Time (min.)
LC_FF0490B
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Food Contaminants Pesticides on Pinnacle® DB Aqueous C18 (LC/MS/MS, ESI-)
NEW! Peak List: Compound bentazone bromoxynil Ioxynil fludioxinil boscalid prothioconazole fipronil noviflumuron
Polarity negative negative negative negative negative negative negative negative
Mol. Wt. 240.28 276.91 370.92 248.19 343.21 344.26 437.15 529.1
RT (min.) 0.77 1.32 2.46 5.34 5.36 5.41 5.56 7.08
Sample: multicomponent pesticide standard Inj.: 5µL Conc.: 33.3ppb each pesticide Sample diluent: acetonitrile
Transition 1 239.0 → 131.8 275.9 → 80.8 369.8 → 126.7 247.0 → 179.9 343.0 → 307 341.9 → 305.8 434.9 → 329.8 527.1 → 343.8
Column: Cat.#: Dimensions: Particle size: Pore size:
Pinnacle® DB Aqueous C18 9418252 50mm x 2.1mm 1.9µm 140Å
Conditions: Mobile phase:
A: 10 mM NH4OAc in water B: 10 mM NH4OAc in methanol Time (min.) 0.0 1.0 8.0 10.0 11.0
Flow: Temp.: Instrument: Det.:
%B 10 10 90 90 10
600µL/min 35°C Shimadzu Prominence® UFLCXR Applied Biosystems 4000 QTRAP® LC/MS/MS system Ion Source: TurboIonSpray®, ESIIonSpray Voltage, ESI voltage: -4.2kV (ESI-) Gas 1: 40psi Gas 2: 60psi Source Temp.: 500°C; max pressure ~7,200psi
LC_FF0479B
Sudan Dyes on Ultra Aqueous C18 Fixed Dual Wavelength Detector
1. 2. 3. 4.
Sudan I Sudan II Sudan III Sudan IV
wavelength change
Photodiode Array Detector
LC_FF0326
Monitor Sudan I, II, III, and IV in a single, isocratic analysis!
LC_FF0327
Sample: Inj.: 20µL, mixture of Sudan I, Sudan II, Sudan III, Sudan IV Conc.: 20µg/mL each component Sample diluent: methanol Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra Aqueous C18 9178565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase: Flow: Temp.: Det.:
methanol:water, 97:3 1mL/min. ambient UV @ 476/493/512/357nm (left) or UV @ 488/520nm (right)
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HPLC CHROMATOGRAMS | FOODS & FLAVORS
Drug Residues Antibiotic Fluoroquinolones on Allure® PFP Propyl
Sample: Inj.: 5µL Conc.: ~50µg/mL each component Sample diluent: mobile phase
Peak List: 1. norfloxacin 2. levofloxacin 3. ciprofloxacin 4. lomefloxacin 5. enrofloxacin 6. sparfloxacin
Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase:
High organic (60% acetonitrile) means better LC/MS sensitivity!
Flow: Temp.: Det.:
Allure® PFP Propyl 9169565 150mm x 4.6mm 5µm 60Å 10mM potassium phosphate monobasic (pH 2.5): acetonitrile, 40:60 (v/v) 1.0mL/min. ambient UV @ 220nm
LC_PH0420
Nonsteroidal Anti-inflammatory Drugs (NSAIDs) Ultra II® Biphenyl
NEW! Peak List 1. ketoprofen 2. piroxicam 3. sulindac 4. diclofenac
Sample: Inj.: 5µL Conc.: 15µg/mL each component Sample diluent: methanol Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Instrument: Mobile phase: Flow: Temp.: Det.:
Ret. Time (min.) 0.894 1.068 1.197 1.465
Ultra II® Biphenyl 9609853 50mm x 3.0mm 2.2µm 100Å Shimadzu Prominence UFLCXR 20mM potassium phosphate, pH 2.5 with H3PO4:methanol (30:70) 1.2mL/min. 40°C UV @ 254nm
LC_PH0487
For more information on Biphenyl columns, visit
www.restek/biphenyl
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HPLC CHROMATOGRAMS | FOODS & FLAVORS
Drug Residues Nitrofuran Metabolites on Ultra C18 (LC/MS/MS)
d5-AMOZ
AMOZ
AHD
d4-AOZ
AOZ
SC
LC_0323
Peak List: 1. d5-AMOZ 2. AMOZ 3. AHD 4. d4-AOZ 5. AOZ 6. SC Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase:
Flow: Temp.: Det.:
Conc. (ppb) 0.3 0.3 0.3 0.3 0.3 0.3
Time (min) 3.7 3.8 5.46 5.49 5.51 5.6
Ultra C18 9174312 100mm x 2.1mm 3µm 100Å A: 0.05% formic acid in methanol B: 0.05% formic acid – 5 mM NH4 formate in water Time (min) %B 0 90 2.5 90 5 10 10 10 12 90 15 90 200µL/min. 30°C MS/MS triple quadrupoles (Thermo Scientific Discovery)
Analyzer Parameters: Ion source: ESI (electrospray ionization) Only segment: 15 min. Polarity: positive Data type: centroid Scan mode: SRM product Scan width (m/z): 0.7 Scan time (s): 0.25 Peak width: Q1: within 0.7 Q2: 0.7 Collision gas pressure (mTorr): 1.5 (argon) Divert valve: active, with 3 positions Positions-1° 2 min., 2° 8 min., 3° 5 min. Analyte AOZ AMOZ SC AHD
Prec. Ion 236 335 209 249
Prod. Ion 134 291 166 134
Collision E 12 V 10 V 12 V 12 V
Tube Lens 120 100 80 110
AMOZ = 3-amino-5-morpholinomethyl-2-oxazolidinone AHD = 1-aminohydantoin hydrochloride AOZ = 3-amino-2-oxazolidinone SC = semicarbazide
for more info Visit www.restek.com/cat004 to view our "Analysis of Nitrofurans in Honey Using LC/MS/MS and an Ultra C18 Column" article.
724 www.restek.com Website : www.chromtech.net.au E-mail :
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HPLC CHROMATOGRAMS | FOODS & FLAVORS
Drug Residues Sulfonamides in Milk on Pinnacle® DB Biphenyl (LC-TOFMS)
NEW! Sample:
milk spiked at 200ppb with sulfonamides, extracted with Q-sep™ Q110 QuEChERS extraction tube (cat.# 26213), and cleaned up with Q-sep™ Q211 QuEChERS dSPE clean-up tube (cat.# 26216), then concentrated. Inj.: 10µL Conc.: 4µg/mL Sample diluent: 0.2% formic acid Column: Cat. #: Dimensions: Particle size: Pore size:
Compound 1. sulfadiazine 2. sulfathiazole 3. sulfamerazine 4. sulfamethazine 5. sulfamethoxypyridazine 6. sulfamethoxazole 7. sulfachloropyridazine 8. sulfadimethoxine 9. sulfaquinoxaline
Pinnacle® DB Biphenyl 9409312 100mm x 2.1mm 3µm 140Å
Conditions: Instrument: Mobile phase:
Agilent 1100 with LECO Unique® LC-TOFMS A: 0.2% formic acid B: methanol:acetonitrile (80:20), with 0.2% formic acid Time (min.) %B 0 15 20 100 30 100 Flow: 1mL/min. with preinjection split delivering 0.2mL/min. to column Temp.: ambient Det.: LECO Unique® LC-TOFMS Interface: ESI Ion mode: positive Temp.: 120°C Capillary: 3.8kV Cone: 50V Desolvation gas: 4L/min. Collected: 200-1,000 m/z
[M+H]+ 251 256 265 279 281 254 285 311 301
LC_FF0502
Sulfa Drugs Ultra II® Biphenyl
Peak List Ret. Time (min.) 1. sulfadiazine 1.496 2 sulfathiazole 1.663 3. sulfamerazine 1.997 4. sulfamethoxazole 2.381 5. sulfamethazine 2.523 6. sulfachlorpyridizine 2.651 7. sulfamethoxine 3.228
Sample: Inj.: 3µL Conc.: 25µg/mL each component Sample diluent: methanol Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Instrument: Mobile phase:
Flow: Temp.: Det.:
NEW!
Ultra II® Biphenyl 9609853 50mm x 3.0mm 2.2µm 100Å Shimadzu Prominence UFLCXR 0.1% formic acid in water:methanol (75:25) Time (min.) %B 0 25 3.5 65 1.0mL/min. 30°C UV @ 254nm
LC_PH0489
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HPLC CHROMATOGRAMS | FOODS & FLAVORS
Preser vatives
Innovative food solutions at your fingertips! Visit Restek’s new website at
www.restek.com/fff for all your testing needs.
Carboxylic Acids (Pyruvic and Lactic) on Ultra Aqueous C18 Peak List: 1. pyruvic acid 2. lactic acid Sample: Inj.: Solvent: Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase: Flow: Temp.: Det.:
Conc. (mg/mL) 5 50 5µL 50mM potassium phosphate, pH 2.5
restek innovation!
Ultra Aqueous C18 9178565 150mm x 4.6mm 5µm 100Å
Excellent peak symmetry in 100% aqueous mobile phase!
50mM potassium phosphate, pH 2.5 1.0mL/min. 26°C UV @ 210nm
Carboxylic Acids on Ultra Aqueous C18
Peak List: 1. malonic acid 2. lactic acid 3. acetic acid 4. citric acid 5. succinic acid 6. fumaric acid
4
5 3 1
Sample: Inj.: Solvent:
10µL water
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra Aqueous C18 9178565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase: Flow: Temp.: Det.:
0
LC_0297
Conc. (µg/mL): 500 500 1,000 1,000 2,000 10
6
2
50mM potassium phosphate, pH 2.5: acetonitrile (99:1) 1.5mL/min. 25°C UV @ 210nm
1
2
3
4 min.
LC_0140
726 www.restek.com Website : www.chromtech.net.au E-mail :
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Page 727
HPLC CHROMATOGRAMS | FOODS & FLAVORS
Preser vatives, Sugars Phenolic Antioxidants on Pinnacle® II C18 Peak List: Conc. (ppm) 1. propyl gallate 168 2. TBHQ 182 3. 2-BHA + 3-BHA 197 4. BHT 193
Sample: Inj.: Solvent:
10µL methanol
Column: Cat.#: Dimensions: Particle size: Pore size:
Pinnacle® II C18 9214565 150mm x 4.6mm 5µm 110Å
Conditions: Mobile phase:
1
A: 1% acetic acid B: methanol Time B (min.) (%) 0 50 4 50 10 90 25 90 26 50 1.0mL/min. 30°C UV @ 280nm
3
Flow: Temp.: Det.:
2
4
Topical and timely insights from top chromatographers.
Visit us at blog.restek.com
4
2
0
ChromaBLOGraphy
6
8
10
12
14
16
18 min.
LC_0198
Sugars on Pinnacle® II Amino (3μm)
Peak List: 1. fructose 2. glucose 3. sucrose 4. maltose 5. lactose
Conc. (mg/mL) 2.0 2.1 4.0 4.5 4.4
Sample: Inj.: Conc.:
Sample: Inj.: Solvent:
5µL mobile phase
Column: Cat.#: Dimensions: Particle size: Pore size:
Pinnacle® II Amino 9217365 150mm x 4.6mm 3µm 110Å
Conditions: Mobile phase: Flow: Temp.: Det.:
LC_0223
Vanilla Bean Extract on Pinnacle® DB C18 (Fast LC)
Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase:
water:acetonitrile (25:75, v/v) 1.5mL/min. 35°C refractive index @ 35°C
Flow: Temp.: Det.:
5µL 20% v/v commercial extract in water
Peak List 1. vanillin
Ret. Time (min.) 2.00
Pinnacle® DB C18 9414555 50mm x 4.6mm 5µm 140Å 0.2% v/v phosphoric acid in water:methanol (80:20 v/v) 2mL/min. 27°C UV @ 254nm
LC_0252
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HPLC CHROMATOGRAMS | FOODS & FLAVORS
Flavor Compounds Vanillin on Ultra C8 Sample: Inj.: Conc.:
Vanillin and Ethyl Vanillin on Ultra C8 Peak List: 1. vanillin
Solvent:
10mL 5% solution of vanilla extract 40% ethanol
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra C8 9103565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phases: A: 1.2% acetic acid in water B: methanol Time (min.) %B 0.0 20.0 5.0 20.0 15.0 40.0 20.0 40.0 25.0 20.0 Flow: 1.0mL/min. Temp.: 28°C Det.: UV @ 254nm
0
2
4
6
8
10
12
Peak List: 1. vanillin 2. ethyl vanillin
1
Conc. (mg/mL) 0.12 0.04
Sample: Inj.: Solvent:
10µL 40% ethanol
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra C8 9103565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phases: A: 1.2% acetic acid B: methanol Time (min.) %B 0.0 20.0 5.0 20.0 15.0 40.0 20.0 40.0 25.0 20.0 Flow: 1.0mL/min. Temp.: 28°C Det.: UV @ 254nm
14
16
18 min.
0
2
4
6
LC_0149
12
14
16
18 min.
Peak List: 1. ethanol 2. unknown 3. capsaicin 4. dihydrocapsaicin
Sample: Inj.: Conc.: Solvent:
5µL 100µg/mL methanol
Column: Cat.#: Dimensions: Particle size: Pore size:
Pinnacle II Cyano 9216565 150mm x 4.6mm 5µm 110Å
Conditions: Mobile phase: Flow: Temp.: Det.:
water:acetonitrile (50:50 v/v) 1.0mL/min. ambient UV/Vis @ 345nm
®
Sample: Inj.: Conc.: Solvent: Column: Cat.#: Dimensions: Particle size: Pore size:
1
Conditions: Mobile phase:
Flow: Temp.: Det.:
2
10
Capsaicinoids: Heat Level Assay for Habañero Nuggets on Pinnacle® II C18
Peak List: 1. piperine
1
8
2
LC_0148
Black Pepper: Piperine on Pinnacle® II Cyano
0
1
3
4 LC_0181
5
6
10µL habañero nugget extract 0.8g sample in 200mL ethanol Pinnacle® II C18 9214565 150mm x 4.6mm 5µm 110Å A: 1% acetic acid in water (v/v) B: acetonitrile Time (min.) % B 0 50 2 50 9 80 12 80 13 50 1.0mL/min. ambient UV @ 280nm
7 min. LC_0160
728 www.restek.com Website : www.chromtech.net.au E-mail :
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HPLC CHROMATOGRAMS | FOODS & FLAVORS
Dietar y Supplements Melatonin Ultra II® Aqueous C18
NEW! Peak List 1. melatonin
Ret. Time (min.) 3.117
Sample: Inj.: 1µL Conc.: 100µg/mL Sample diluent: water:methanol (60:40) Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra II® Aqueous C18 9608565 150mm x 4.6mm 5µm 100Å
Conditions: Instrument: Mobile phase: Flow: Temp.: Det.: Pressure:
Shimadzu Prominence UFLCXR water:methanol (60:40) 2.0mL/min. 40°C UV @ 222nm 240bar
LC_PH0506
Organic Acids Ultra II® Aqueous C18
Peak List 1. tartaric acid 2. quinic acid 3. malic acid 4. citric acid 5. fumaric acid
Ret. Time (min.) 2.061 2.210 2.596 4.177 5.853
NEW!
Sample: Inj.: Conc.:
5µL 10µg/mL fumaric acid, 2,000µg/mL each other acids Sample diluent: water Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra II® Aqueous C18 9608565 150mm x 4.6mm 5µm 100Å
Conditions: Instrument: Mobile phase: Flow: Temp.: Det.:
Shimadzu Prominence UFLCXR 100% 20mM potassium phosphate (pH2.5) 1.0mL/min. 30°C UV @ 226nm
LC_PH0498
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HPLC CHROMATOGRAMS | FOODS & FLAVORS
Dietar y Supplements Allicin in Garlic Tablets on Pinnacle® II C18 Sample: Inj.: Conc.:
Peak List: 1. allicin
Phenethyl Glucosinolate on Ultra Aqueous C18
Solvent:
20µL 1.2mg/mL garlic tablet in water deionized water
Column: Cat.#: Dimensions: Particle size: Pore size:
Pinnacle® II C18 9214565 150mm x 4.6mm 5µm 110Å
Conditions: Mobile phase: Flow: Temp.: Det.:
Peak List: 1. phenethyl glucosinolate
water:methanol (50:50 v/v) 0.8mL/min. 30°C UV @ 240nm
Sample: Inj.: Conc.: Solvent:
10µL 1,000µg/mL water
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra Aqueous C18 9178565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase: A: 50mM potassium phosphate, pH 2.5 B: acetonitrile Time (min.): % B 0.0 0 10 75 11 0 16 0 Flow: 1.0mL/min. Temp.: ambient Det.: UV @ 210nm
1
0
2
4
6
8
min. LC_0166
LC_0174
Phenolics in Echinacea on Pinnacle® II C18 Sample: Inj.: Conc.: Solvent: Sample temp.:
10µL 6.7mg/mL Echinacea capsule contents in sample diluent ethanol:water (70:30, v/v) 25°C
Column: Cat.#: Dimensions: Particle size: Pore size:
Pinnacle® II C18 9214565 150mm x 4.6mm 5µm 110Å
Conditions: Mobile phase:
Flow: Temp.: Det.:
Tocopherols in Soy Oil on Pinnacle® II Silica
Peak List: 1. caftaric acid 2. chlorogenic acid 3. echinacoside 4. cichoric acid
Peak List: 1. α-tocopherol 2. β-tocopherol 3. γ-tocopherol 4. δ-tocopherol Sample: Inj.: Conc.: Solvent:
10µL approx. 1.25% soy oil hexane
Column: Cat.#: Dimensions: Particle size: Pore size:
A: 0.1% phosphoric acid B: acetonitrile Time (min.) %B 0 10 13 22 14 40 14.5 40 15 10 20 10 1.5mL/min. 35°C UV @ 330nm
3
Pinnacle® II Silica 9210565 150mm x 4.6mm 5µm 110Å
Conditions: Mobile phase:
isopropyl alcohol:hexane (0.5:99.5) 0.6mL/min. 30°C UV @ 295nm
Flow: Temp.: Det.:
4 1 2
0 LC_0195
2
4
6
8
10
12
14
min.
LC_0197
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HPLC CHROMATOGRAMS | FOODS & FLAVORS
Vitamins & Dietar y Supplements Vitamin D on Ultra Aqueous C18 (LC/MS/MS)
NEW!
Sample: Inj.: Conc.:
20µL extracted serum sample
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra Aqueous C18 9178352 50mm x 2.1mm 3µm 100Å
Conditions: Instrument: Mobile phase:
Flow: Temp.: Det.: Ion source: Mode: Dwell time: Compound d6-25-OH D3 (IS) 25-OH D3 25-OH D2 25-OH D3 25-OH D2 25-OH D2
Shimadzu Prominence® UFLCXR A: 0.1% formic acid in water B: 0.1% formic acid in methanol Time (min.) %B 0.00 50 2.5 100 3.5 100 3.6 50 5.00 50 700µL/min. 40°C Applied Biosystems 3200 QTRAP™ LC/MS/MS system TurboIonSpray®, APCI+ MRM 100msec. Q1 389.3 383.3 395.3 383.3 395.3 395.3
Q3 211.2 211.2 229.2 229.2 269.2 119.0
Declustering potential (V) 68 68 55 68 55 55
Ret. Time (min.) 3.00 3.00 3.04 3.00 3.04 3.04
LC_CF0492
for more info Visit www.restek.com/cat003 to view our "5 Minute Analysis of Vitamin D in Serum by LC/MS/MS" article.
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HPLC CHROMATOGRAMS | FOODS & FLAVORS
Vitamins & Dietar y Supplements Vitamins (Fat Soluble) on Ultra C18 Peak List: Conc. (mg/mL) 1. solvent front n/a 2. menadione (vitamin K3) 0.45 3. all-trans-retinol (vitamin A) 0.34 4. vitamin D3 0.4 5. unknown n/a 6. α tocopherol (vitamin E) 2.4 7. α tocopherol acetate (vitamin E acetate) 2.4 8. unknown n/a 9. phylloquinone (vitamin K1) 0.84
Vitamins Thiamin (Vitamin B1) and Ascorbic Acid (Vitamin C) on Ultra IBD
Sample: Solvent:
diethyl ether
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra C18 9174575 250mm x 4.6mm 5µm 100Å
Conditions: Mobile phase:
2
4
Temp.: Det.:
Peak List: Conc. (mg/mL) 1. thiamin 2.5 2. ascorbic acid 2.5
acetonitrile:methanol (90:10, v/v) Time (min.) Flow (mL/min.) 1.00 1.00 5.00 1.00 5.01 2.00 50.0 2.00 30°C UV @ 280nm
Sample: Inj.: Solvent:
1.0µL mobile phase
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra IBD 9175565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase: Flow: Temp.: Det.:
50mM potassium phosphate, pH 2.5 1.0mL/min. ambient UV @ 254nm
3 6 7 9
1 5
0
10
20
8
30 LC_0144
40
min. LC_0227
Restek is your Complete Source for
Analyzing Pesticides in Dietary Supplements For more information on dietary supplements, visit
www.restek.com/dietary
732 www.restek.com Website : www.chromtech.net.au E-mail :
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Page 733
HPLC CHROMATOGRAMS | FOODS & FLAVORS
Vitamins & Dietar y Supplements Fat Soluble Vitamins Ultra II® C18
NEW! Peak List 1. vitamin K2 2. vitamin D2 3. vitamin D3 4. vitamin E acetate 5. vitamin K1
Ret. Time (min.) 0.905 1.224 1.305 1.828 2.169
Sample: Inj.: 1µL Conc.: 100µg/mL each component Sample diluent: acetone
1
2
3
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra II® C18 9604853 50mm x 3.0mm 2.2µm 100Å
Conditions: Instrument: Mobile phase: Flow: Temp.: Det.:
Shimadzu Prominence UFLCXR acetonitrile:methanol (85:15) 1.5mL/min. ambient UV @ 230nm
5 4
0.00
0.25
0.50
0.75
1.00
1.25
1.50
1.75
2.00
2.25
2.50
min.
LC_PH0492
Vitamins B1, B2, B3, and B6 Ultra C8 Peak List 1. vitamin B3 (nicotinamide) 2. vitamin B6 (pyridoxine) 3. vitamin B2 (riboflavin) 4. vitamin B1 (thiamine)
NEW!
Ret. Time (min.) 2.732 5.615 8.719 19.004
Sample: Inj.: 5µL Conc.: 50µg/mL each component Sample diluent: water:methanol (80:20) Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Instrument: Mobile phase: Flow: Temp.: Det.:
Ultra C8 9103575 250mm x 4.6mm 5µm 100Å Shimadzu Prominence UFLCXR 0.008M sodium 1-hexanesulfonate:methanol:glacial acetic acid: triethylamine (817.5:175:7.5:0.2) 2.0mL/min. 50°C UV @ 270nm
LC_PH0497
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Page 734
HPLC CHROMATOGRAMS | FOODS & FLAVORS
Vitamins & Dietar y Supplements Water Soluble B Vitamins Ultra II® Aqueous C18
NEW!
Sample: Inj.: 1µL Conc.: 100µg/mL each component Sample diluent: water:methanol (70:30) Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Instrument: Mobile phase:
Flow: Temp.: Det.: Pressure:
Ultra II® Aqueous C18 9608565 150mm x 4.6mm 5µm 100Å
Peak List 1. vitamin B3 2. vitamin B6 3. vitamin B5 4. vitamin B9 5. vitamin B12 6. vitamin B2 7. vitamin B7
Ret. Time (min.) 2.472 2.764 3.804 4.126 4.261 4.329 4.472
Shimadzu Prominence UFLCXR A. 10mM potassium phosphate (pH 2.5) B. acetonitrile Time (min.) %B 0 2 1.5 2 5 75 1.5mL/min. 40°C UV @ 210nm 105bar
LC_PH0503
Vitamin C Ultra II® Aqueous C18
NEW!
Peak List Ret. Time (min.) 1. ascorbic acid (vitamin C) 1.759 Sample: Inj.: 2µL Conc.: 100µg/mL Sample diluent: water Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra II® Aqueous C18 9608565 150mm x 4.6mm 5µm 100Å
Conditions: Instrument: Mobile phase: Flow: Temp.: Det.: Pressure:
Shimadzu Prominence UFLCXR 100% 10mM potassium phosphate (pH2.5) 1.5mL/min. 40°C UV @ 242nm 95bar
LC_PH0504
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Page 735
HPLC CHROMATOGRAMS | FOODS & FLAVORS
Vitamins & Dietar y Supplements Vitamin A Compounds Ultra II® Aromax
NEW! Peak List 1. retinol 2. β-carotene
Ret. Time (min.) 0.832 0.950
Sample: Inj.: 2µL Conc.: 100µg/mL each component Sample diluent: acetone
UVat 325 (in red) UVat 480 (in black)
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra II® Aromax 9607853 50mm x 3.0mm 2.2µm 100Å
Conditions: Instrument: Mobile phase: Flow: Temp.: Det.: Pressure:
Shimadzu Prominence UFLCXR methanol:acetonitrile (99:1) 0.8mL/min. ambient UV @ 325nm and 480nm 105bar
LC_PH0505
Chromatogram Search Tool Search by compound name, synonym, CAS # or keyword
www.restek.com/chromatograms
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HPLC CHROMATOGRAMS | PHARMACEUTICAL
Famotidine Famotidine and USP Impurities Ultra II® Aromax
Sample: Inj.: Conc.:
Fast! 5μm
Column: Cat.#: Dimensions: Particle size: Pore size:
Peak List 1. impurity A 2. famotidine 3. impurity D 4. impurity C 5. impurity B
NEW!
10µL 100µg/mL famotidine, 10µg/mL each impurity Sample diluent: methanol
Ret. Time (min.) 3.779 8.264 9.180 9.911 14.018
Conditions: Instrument: Mobile phase:
Flow: Temp.: Pressure: Det.:
Ultra II® Aromax 9607565 150mm x 4.6mm 5µm 100Å Shimadzu Prominence UFLCXR 20mM potassium phosphate (pH 2.5):methanol Time (min.) %B 0 5 15 35 2.0mL/min. 40°C 140bar UV @ 268nm
15 min.
LC_PH0495
Sample: Inj.: 5µL Conc.: 100µg/mL famotidine, 10µg/mL each impurity Sample diluent: methanol
Faster! 3μm Peak List 1. impurity A 2. famotidine 3. impurity D 4. impurity C 5. impurity B
Column: Cat.#: Dimensions: Particle size: Pore size:
Ret. Time (min.) 2.094 4.460 4.937 5.334 7.480
Conditions: Instrument: Mobile phase:
Flow: Temp.: Pressure: Det.:
Ultra II® Aromax 9607313 100mm x 3.2mm 3µm 100Å Shimadzu Prominence UFLCXR 20mM potassium phosphate (pH 2.5):methanol Time (min.) %B 0 5 8 35 1.2mL/min. 40°C 345bar UV @ 268nm
8 min.
LC_PH0494
Sample: Inj.: 2µL Conc.: 100µg/mL famotidine, 10µg/mL each impurity Sample diluent: methanol
Fastest! 2.2μm Peak List 1. impurity A 2. famotidine 3. impurity D 4. impurity C 5. impurity B
Column: Cat.#: Dimensions: Particle size: Pore size:
Ret. Time (min.) 0.892 2.082 2.292 2.517 3.540
Conditions: Instrument: Mobile phase:
Flow: Temp.: Pressure: Det.:
Ultra II® Aromax 9607853 50mm x 3.0mm 2.2µm 100Å Shimadzu Prominence UFLCXR 20mM potassium phosphate (pH 2.5):methanol Time (min.) %B 0 5 4 35 1.2mL/min. 40°C 480bar UV @ 268nm
4 min.
LC_PH0493
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HPLC CHROMATOGRAMS | PHARMACEUTICAL
Potential Genotoxic Impurities, Analgesics Potential Genotoxic Impurities Ultra II® Aromax
NEW!
Peak List 1. ethyl benzenesulfonate 2. isopropyl toluenesulfonate 3. n-propyl toluenesulfonate 4. n-butyl toluenesulfonate *All M/Z are [M+NH4]+
M/Z* 204 232 232 246
Sample: Inj.: 100µL Conc.: 10ng/mL each PGI in 1mg/mL dorzolamide Sample diluent: mobile phase Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Instrument: Mobile phase: Flow: Temp.: Det.:
Ultra II® Aromax 9607312 100mm x 2.1mm 3µm 100Å Agilent 1100 A: 0.2% ammonium acetate in water B: 0.2% ammonium acetate in methanol 0.3mL/min. ambient LECO TOF-MS
LC_PH0482B
Hydroxybenzoic Acids on Ultra Aqueous C18
1. 2. 3. 4. 5. 6. 7.
3,5-dihydroxybenzoic acid 2,5-dihydroxybenzoic acid 4-hydroxybenzoic acid 3-hydroxybenzoic acid 2,4-dihydroxybenzoic acid benzoic acid salicylic acid
Sample: Inj.: 5µL Conc.: 50µg/mL each component Sample diluent: mobile phase Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase:
Flow: Temp.: Det.:
Ultra Aqueous C18 9178565 150mm x 4.6mm 5µm 100Å A: 20mM potassium phosphate (pH 2.5); B: acetonitrile Time (min.) %B 0.00 20 5.00 50 10.00 50 1.0mL/min. ambient UV @ 210nm
LC_PH0400
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HPLC CHROMATOGRAMS | PHARMACEUTICAL
Analgesics Analgesic Acetaminophen and Narcotic Analgesics on Ultra C18 Peak List: Conc. (µg/mL) U. unknown unknown 1. morphine sulfate 204 2. acetaminophen 92 3. codeine phosphate 216 4. oxycodone HCl 206 5. hydrocodone bitartrate 218
Sample: Inj.: Sample: Solvent:
4.0µL raw material mix mobile phase
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra C18 9174575 250mm x 4.6mm 5µm 100Å
Narcotic Analgesic Morphine Sulfate Raw Material USP 25: Resolution Solution on Pinnacle® II C18 Peak List: Conc. (µg/mL) 1. unknown unknown 2. phenol 0.15 3. morphine sulfate 0.24 Sample: Inj.: Solvent:
Conditions: Mobile phase:
Flow: Temp.: Det.:
A: 10mm potassium phosphate, pH 2.8 B: acetonitrile:methanol, (90:10, v/v) (85A:15B, v/v) 1.0mL/min. 35°C UV @ 235nm
20µL mobile phase
Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase:
Flow: Temp.: Det.:
Pinnacle® II C18 9214575-700 250mm x 4.6mm 5µm 110Å A: 0.73g 1-heptane sulfonic acid and 10mL glacial acetic acid, diluted to 720mL with water, pH = 2.33 B: methanol (72A:28B, v/v) 1.00mL/min. 26.5°C UV @ 284nm
LC_0209
LC_0221
Narcotic Analgesic Hydrocodone Bitartrate by USP 25 on Pinnacle® II Silica Peak List: 1. hydrocodone bitartrate Sample: Inj.: Conc.: Sample: Solvent:
Ret. Time (min.) 3.1
Inj. % RSD (n = 5) 0.37%
20µL 1µg/mL assay standard mobile phase
Column: Cat.#: Dimensions: Particle size: Pore size:
Flow: Temp.: Det.:
Peak List: Conc. (µg/mL) 1. codeine phosphate 13 2. oxycodone HCl 9 Sample: Inj.: Sample: Solvent:
Pinnacle® II Silica 9210575 250mm x 4.6mm 5µm 110Å
Conditions: Mobile phase:
Narcotic Analgesic Oxycodone HCl Raw Material by USP 25: Resolution Solution on Ultra C8
Conditions: Mobile phase:
Flow: Temp.: Det.:
RRT 0.7 1.0
Tailing 1.1 0.99
Res. — 3.3
10µL resolution solution mobile phase
Column: Cat.#: Dimensions: Particle size: Pore size:
A: 800mL acetonitrile; 4mL water; 1mL diethylamine B: methanol (55A:45B, v/v) 1.5mL/min. 27°C UV @ 280nm
Ret. Time (min.) 19.3 26.8
Ultra C8 9103564 150mm x 4.0mm 5µm 100Å A: 5mM hexane sulfonic acid with 2mL TEA and 3mL of phosphoric acid per liter. pH adjusted to 2.5 B: methanol (90A:10B, v/v) 1.5mL/min. 40°C UV @ 206nm
Impurities resolved from primary peak.
0
1
2
3
4
min.
LC_0211
LC_0213
738 www.restek.com Website : www.chromtech.net.au E-mail :
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HPLC CHROMATOGRAMS | PHARMACEUTICAL
Antiarrhythmics, Antiasthmatics Antiarrhythmic Calcium Channel Blockers on Ultra Cyano Sample: Inj.: Conc.: Solvent:
Peak List: 1. diltiazem 2. nifedipine impurity 3. verapamil 4. nifedipine 5. nicardipine
Peak List: 1. uracil (marker) 2. furosemide
5µL 100mg/mL acetonitrile:water (1:1)
Column: Cat.#: Dimensions: Particle size: Pore size:
Sample: Inj.: Conc.: Solvent:
Ultra Cyano 9106565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase: 1
Antiarrhythmic Furosemide on Ultra IBD
20mM potassium phosphate monobasic, pH 3.0:acetonitrile (70:30, v/v) 1.2mL/min. 30°C (±1°C) 235nm
Flow rate: Temp.: Det.:
Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase:
Flow rate: Temp.: Det.:
4
5µL 1000µg/mL acetonitrile:water (60:40, v/v)
Impurities in Furosemide (enlargement of baseline)
Ultra IBD 9175565 150mm x 4.6mm 5µm 100Å 20mM ammonium acetate, pH 4.5: acetonitrile (70:30, v/v) 2.0mL/min. 27°C UV @ 280nm
5 3
2
0
1
2
3
4
5
6
7
8
9 min.
LC_0062
LC_0071
Antiasthmatic Beclomethasone on Ultra C18
Peak List: 1. beclomethasone 2. deschlorobeclomethasone 1
Sample: Inj.: Conc.: Solvent:
10µL 79.5µg/mL (beclomethasone) water:acetonitrile (10:90, v/v)
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra C18 9174565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase: Flow: Temp.: Det.:
water:acetonitrile (70:30, v/v) 1.0mL/min. 25°C UV @ 238nm
Chromatogram Search Tool Search by compound name, synonym, CAS # or keyword
www.restek.com/chromatograms
2
0
5
10
15
20 min.
LC_0120
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Page 740
HPLC CHROMATOGRAMS | PHARMACEUTICAL
Antibiotics Antibiotic Cephalexin on Ultra IBD
Antibiotic Penicillin V on Ultra IBD
1 1
Peak List: 1. penicillin V
Peak List: 1. cephalexin 2. cephradine Sample: Inj.: Conc.: Solvent:
Sample: Inj.: Conc.: Solvent:
10µL 500µg/mL acetonitrile:water (10:90, v/v)
Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase:
Flow rate: Temp.: Det.:
2
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra IBD 9175565 150mm x 4.6mm 5µm 100Å 10mM ammonium formate, pH 2.5: acetonitrile (90:10, v/v) 1.2mL/min. 30°C UV @ 270nm
4
6
8
4
5
6
7
Peak List 1. oxytetracycline 2. degradation peak 3. tetracycline 4. degradation peak 5. chlortetracycline 6. doxycycline
Peak List: 1. cephaloridine Sample: Inj.: Conc.: Solvent:
8
min.
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra IBD 9175565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase:
Conditions: Mobile phase:
acetonitrile: 20mM ammonium phosphate, pH 4.0 (20:80, v/v) 1.2mL/min. 27°C UV @ 254nm
Flow: Temp.: Det.:
6
7
8
Ret. Time (min.) 4.59 5.19 6.32 11.16 15.81 19.66
Sample: Inj.: 30µL Conc.: 100µg/mL each component Sample diluent: methanol
5µL 1mg/mL acetonitrile:water (50:50, v/v)
Column: Cat.#: Dimensions: Particle size: Pore size:
5
3
Tetracycline Mix on Allure® Biphenyl
1
4
2
LC_0096
Antibiotic Cephaloridine on Ultra IBD (Reversed Phase)
3
1
10 min.
LC_0094
2
10mM ammonium formate, pH 2.5: acetonitrile (95:5, v/v) 1.2mL/min. 30°C UV @ 270nm
Flow rate: Temp.: Det.:
0 2
1
Ultra IBD 9175565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase:
0
0
2.5µL 1.2mg/mL acetonitrile:water (10:90,v/v)
Flow: Temp.: Det.:
Allure® Biphenyl 9166565 150mm x 4.6mm 5µm 60Å 20mM ammonium phosphate (pH 2.5):acetonitrile, 80:20 1mL/min. ambient UV @ 254nm
min.
LC_PH0347
LC_0102
740 www.restek.com Website : www.chromtech.net.au E-mail :
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HPLC CHROMATOGRAMS | PHARMACEUTICAL
Antibiotics, Anti-Diabetic, Cor ticosteroids Antibiotic Amoxicillin on Ultra IBD
Anti-Diabetic Glyburide (Glybenclamide) on Ultra IBD 2
1
Peak List: 1. amoxicillin Sample: Inj.: Conc.: Solvent:
Peak List: 1. unknown 2. glyburide
5µL 1.5mg/mL acetonitrile:water (10:90, v/v)
Column: Cat.#: Dimensions: Particle size: Pore size:
Sample: Inj.: Conc.: Solvent:
Ultra IBD 9175565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase:
Column: Cat.#: Dimensions: Particle size: Pore size:
10mM ammonium formate, pH 2.5: acetonitrile (95:5, v/v) 1.2mL/min. 30°C UV @ 270nm
Flow rate: Temp.: Det.:
1µL 500µg/mL water:methanol:acetonitrile (1:1:2, v/v)
Conditions: Mobile phase: Flow rate: Temp.: Det.:
Ultra IBD 9175565 150mm x 4.6mm 5µm 100Å water with 0.1% (v/v) acetic acid: acetonitrile (95:05, v/v) 1.0mL/min. 27°C UV @ 254nm
1
0
1
2
3
4
5
6
min. 0
LC_0095
5µL 100µg/mL mobile phase
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra C18 9174565 150mm x 4.6mm 5µm 100Å
Flow: Temp.: Det.:
4
Peak List 1. hydrocortisone 2. prednisone 3. cortisone 4. dexamethasone 5. corticosterone 6. cortisone acetate 7. desoxycorticosterone
5
min.
3
Ret. Time (min.) 4.19 4.79 5.08 6.37 9.01 15.75 25.94
Sample: Inj.: 5µL Conc.: 100µg/mL each component Sample diluent: methanol
5
Conditions: Mobile phase:
3
Corticosteroids on Allure® Biphenyl
2
Sample: Inj.: Conc.: Solvent:
2
LC_0069
Corticosteroids on Ultra C18 Peak List: 1. uracil (void marker) 2. triamcinolone 3. hydrocortisone 4. dexamethasone 5. corticosterone 6. deoxycorticosterone
1
4
6
water:methanol (30:70, v/v) 1.0mL/min. 30°C UV @ 220nm
Column: Cat.#: Dimensions: Particle size: Pore size:
Allure® Biphenyl 9166565 150mm x 4.6mm 5µm 60Å
Conditions: Mobile phase: Flow: Temp.: Det.:
water:acetonitrile, 60:40 1mL/min. ambient UV @ 254nm
1
0
1
2
3
4
5
LC_PH0330
6 min.
LC_0108
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HPLC CHROMATOGRAMS | PHARMACEUTICAL
Steroids Endogenous Steroids on Allure® Biphenyl
Peak List 1. β-estradiol 2. testosterone 3. progesterone
Contraceptive Hormones on Allure® Biphenyl Peak List 1. β-estradiol 2. ethynyl estradiol 3. norethindrone 4. norgestrel
Ret. Time (min.) 3.76 5.08 14.05
Sample: Inj.: 5µL Conc.: 100µg/mL each component Sample diluent: methanol Column: Cat.#: Dimensions: Particle size: Pore size:
Allure® Biphenyl 9166565 150mm x 4.6mm 5µm 60Å
Conditions: Mobile phase: Flow: Temp.: Det.:
water:acetonitrile, 40:60 1mL/min. ambient UV @ 220nm
Ret. Time (min.) 3.80 4.45 6.15 8.41
Sample: Inj.: Conc.:
5µL 100µg/mL each component Sample diluent: methanol Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase: Flow: Temp.: Det.:
Allure® Biphenyl 9166565 150mm x 4.6mm 5µm 60Å water:acetonitrile, 40:60 1mL/min. ambient UV @ 220nm
LC_PH0328 LC_PH0334
Hormones on 1.9μm Pinnacle® DB Biphenyl
Get More!
Peak List: 1. estriol 2. 17β-estradiol 3. 17α-estradiol 4. ethynyl estradiol 5. testosterone 6. estrone 7. norethindrone
Pharmaceutical Solutions Online www.restek.com/pharma
LC_PH0453
Sample: Inj.: 1µL Conc.: 100µg/mL each component Sample diluent: water:methanol (50:50) Column: Cat.#: Dimensions: Particle size: Pore size:
1.9µm Pinnacle® DB Biphenyl 9409252 50mm x 2.1mm 1.9µm 140Å
Conditions: Mobile phase:
Flow: Temp.: Det.:
A: water B: methanol Time(min) %B 0 30 1 30 3 70 0.8 mL/min. 30ºC UV @ 220nm
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HPLC CHROMATOGRAMS | PHARMACEUTICAL
Lactulose Concentrate, Xanthines, Admixed Drugs Laxative Lactulose Concentrate by USP 25: Resolution Solution on Pinnacle® II Amino Peak List: 1. fructose 2. galactose 3. epi-lactose 4. lactulose 5. lactose
Conc. (mg/mL)
Ret. Time (min.)
RRT
Res.
0.4 6.4 unknown 40 4.8
4.6 6.1 10.0 10.5 12.7
0.44 0.58 0.95 1.00 1.21
— — — 1.1 2.9
Xanthines on Pinnacle® DB C18 1
Inj. % RSD (n = 5) — — — 0.3% —
2
Pinnacle® II Amino 9217365 150mm x 4.6mm 3µm 110Å
Conditions: Mobile phase: Flow: Temp.: Det.:
10mM sodium phosphate monobasic in water, pH 4.8: acetonitrile (22:78, v/v) 1.3mL/min. 40°C refractive index @ 40°C
3 4
0
1
2
4
3
5
2µL mobile phase
Column: Cat.#: Dimensions: Particle size: Pore size:
Pinnacle® DB C18 9414565 150mm x 4.6mm 5µm 140Å
Conditions: Mobile phase: 20mM KH2PO4, pH 3.0: acetonitrile (90:10 v/v) Flow: 1.0mL/min. Temp.: 27°C Det.: UV @ 254nm
Sample: Inj.: 20µL resolution standard Solvent: acetonitrile:water (50:50) Column: Cat.#: Dimensions: Particle size: Pore size:
Sample: Inj.: Solvent:
6
7
min.
LC_0241
Peak List: Conc. (µg/mL) 1. theobromine 225 2. theophylline 418 3. β-hydroxyethyltheophylline 418 4. caffeine 400 LC_0222
Admixtures: Hydrocodone Bitartrate/Acetaminophen by USP 25 on Ultra C18 Peak List:
Conc. (µg/mL) 1. acetaminophen 0.76 2. hydrocodone bitartrate 0.0076
Sample: Inj.: Sample:
Ret. Time (min.) 3.304 6.664
RRT 1.0 2.0
Solvent:
25µL 500mg acetaminophen with 5mg hydrocodone bitartrate (tablet) mobile phase
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra C18 9174575 250mm x 4.6mm 5µm 100Å
Conditions: Mobile phase:
Flow: Temp.: Det.:
6.8g potassium monobasic with 0.2mL TEA per liter: acetonitrile (85:15, v/v) 1.5mL/min. 27°C UV @ 210nm (hydrocodone bitartrate) and 295nm (acetaminophen).
Admixtures: Expectorant/Antitussive Guaifenesin and Narcotic Analgesic Codeine on Ultra Phenyl
Tailing Resolution Inj. % RSD (n = 5) 1.1 — 1.1 1.1 17.0 0.9
2 1
Peak List: 1. codeine 2. guaifenesin Sample: Inj.: Solvent:
5µL methanol:water (50:50)
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra Phenyl 9105565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase:
10mM ammonium formate, pH 2.5: acetonitrile (80:20, v/v) 1.2mL/min. 25°C UV @ 280nm
Flow rate: Temp.: Det.:
0 LC_0212
Conc. (µg/mL) 825 375
1
2
3
4
5
6
min.
LC_0088
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HPLC CHROMATOGRAMS | BIOCHEMICAL/CLINICAL
Nucleotides, Nucleosides, Nucleic Acid Bases Nucleotides, Nucleosides, and Nitrogenous Bases on Ultra IBD Sample: Inj.: Solvent:
Peak List: Conc. (µg/mL) 1. CDP 146 2. CMP 113 3. ATP 117 5 4. ADP 145 5. cytidine 155 6. TMP 157 7. AMP 125 8. guanine 36 9. guanosine 103 10. adenine 38 11. adenosine 115
4
6
Uracil, Uridine, UTP, UDP, and UMP on Ultra IBD
Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase: Flow: Temp.: Det.:
7
3
20µL 20mM ammonium acetate, pH 5.8 Ultra IBD 9175565 150mm x 4.6mm 5µm 100Å
Peak List: 1. UTP 2. UDP 3. UMP 4. uracil 5. uridine
5 4
2
Sample: Inj.: Solvent:
20mM ammonium acetate, pH 5.8:methanol (97.5:2.5) 1.0mL/min. 35°C UV @ 260nm
20µL 20mM ammonium acetate, pH 5.8
Column: Cat.#: Dimensions: Particle size: Pore size: 1
Conditions: Mobile phase:
2 1
Flow: Temp.: Det.:
3 11
Conc. (µg/mL) 360 360 284 90 230
Ultra IBD 9175565 150mm x 4.6mm 5µm 100Å 20mM ammonium acetate, pH 5.8: methanol (97.5:2.5) 1.0mL/min. 35°C UV @ 260nm
10 8 9
0
2
4
6
8
10
12
14
16
18
min.
0
2
4 LC_0133
LC_0128
Nucleic Acid Bases on Allure® PFP Propyl
Thymine, Thymidine, TTP, TDP, and TMP on Ultra IBD
Peak List: 1. cytosine 2. uracil 3. guanine 4. thymine 5. adenine
Peak List: Conc. (µg/mL) 1. TTP 224 2. TDP 152 3. TMP 346 4. thymine 88 5. thymidine 318
3
Sample: Inj.: Solvent:
4 5
6 min.
Sample: Inj.: Conc. Solvent:
20µL 20mM ammonium acetate, pH 5.8
Column: Cat.#: Dimensions: Particle size: Pore size:
2 3
Column: Cat.#: Dimensions: Particle size: Pore size:
2
20mM ammonium acetate, pH 5.0: methanol (85:15, v/v) 1.0mL/min. ambient UV @ 260nm
Flow: Temp.: Det.:
20mM ammonium acetate, pH 5.8: methanol (97.5:2.5) 1.0mL/min. 35°C UV @ 260nm
Flow: Temp.: Det.:
1
Allure® PFP Propyl 9169565 150mm x 4.6mm 5µm 60Å
Conditions: Mobile phase:
Ultra IBD 9175565 150mm x 4.6mm 5µm 100Å
Conditions: Mobile phase:
1.0µL 100µg/mL each 20 mM ammonium acetate, pH 5.0 Note: 1-3 drops of ammonium hydroxide added to keep guanine in solution
4
5
1
0
2
4
6
8
10 LC_0132
12
14
16 min.
0
1
2
3
4
5
6
7
8
9 min.
LC_0137
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Page 745
HPLC CHROMATOGRAMS | BIOCHEMICAL/CLINICAL
Catecholamines, Purines/Pyrimidines, Amino Acids, Peptides Catecholamines on Allure® PFP Propyl
Purines/Pyrimidines on Ultra PFP 1
Peak List Retention Time (min.) 1. norepinephrine 3.50 2. levodopa 5.43 3. epinephrine 7.27 4. tyrosine 7.77 5. dopamine 11.3
3
Sample: Inj.: Conc.: Sample diluent: Sample temp.:
10µL 40µg/mL each component mobile phase ambient
Column: Cat.#: Dimensions: Particle size: Pore size:
Allure PFP Propyl 9169565 150mm x 4.6mm 5µm 60Å
Conditions: Mobile phase: Flow: Temp.: Det.:
Peak List: 1. cytosine 2. 5-methylcytosine 3. uracil 4. uric acid 5. hypoxanthine 6. xanthine 7. thymine 8. purine 9. theobromine 10. ethyluracil 11. 6-methylpurine 12. 1,7-dimethylxanthine 13. theophylline 14. propyluracil
5 6
®
7 8
2
50mM ammonium phosphate (pH 3.0, adjusted with formic acid) 1mL/min. 35°C UV @ 266nm
9
Sample: Inj.: Conc.: Solvent:
10
4
10µL 100µg/mL each mobile phase
11 12
0
5
10
15
14
13
20
25 min.
LC_0136
Column: Cat.#: Dimensions: Particle size: Pore size: Conditions: Mobile phase: Flow: Temp.: Det.:
LC_BA0349
Sample: Inj.: Solvent:
20µL mobile phase A
Column: Cat.#: Dimensions: Particle size: Pore size:
Ultra Aqueous C18 9178565 150mm x 4.6mm 5µm 100Å
1. 2. 3. 4. 5. 6. 7.
3
* mfr. D
mfr. C
mfr. F 4 1 2
0.0
4
6 LC_0146
8
10
min.
2.0
4.0
6.0
8.0
10.0
Sample: Inj.:
20µL peptide test mix
Column: Cat.#: Dimensions: Particle size: Pore size:
Viva C18 9514562 150mm x 2.1mm 5µm 300Å
5 6
3
2
Viva
1
2
val-tyr val-tyr-val meth enkephalin val-4-angiotensin III angiotensin II, III leucine enkephalin angiotensin I
mfr. H
Conditions: Mobile phases: A: 50mM potassium phosphate, pH 2.5 B: acetonitrile 5% - 20% B: 0-5 min. 20% - 5% B: 5-6 min. Hold at 5% B: 6-13 min. Flow: 1.0mL/min. Temp.: 30°C Det.: UV @ 254nm
0
0.001% formic acid in water: methanol (50:50, v/v) 1.0mL/min. ambient UV @ 260nm
Peptide Mix on Viva C18
Aromatic Amino Acids on Ultra Aqueous C18 Peak List: Conc. (mg/mL) 1. tyrosine 0.2 2. phenylalanine 1.6 3. tryptophan 0.4
Ultra PFP 9176575 250mm x 4.6mm 5µm 100Å
12.0
14.0
16.0
7 18.0 20.0 LC_BC0386
Conditions: Mobile phase:
A: 0.08% trifluoroacetic acid in water (pH 2.0) B: 0.08% TFA in acetonitrile, 10% B to 40% B in 20 min. Flow: 0.2mL/min. Det.: UV @ 214nm * High levels of trace metals (~100ppm) in the silica enhance this separation.
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HPLC CHROMATOGRAMS | BIOCHEMICAL/CLINICAL
Proteins, Oxytocin Protein Mix on Viva C18
Note excellent peak shapes and resolution on the Viva column! Viva C18 (cat.# 9514562)
Peak List 1. ribonuclease A 2. cytochrome c 3. holo-transferrin 4. apomyoglobin
Manufacturer A
Ret. Time (min.) 11.31 14.65 16.32 20.34
Manufacturer B
Sample: Inj.: 20µL Conc.: 0.08mg/mL each protein Sample diluent: 0.10% TFA in water / 0.10% TFA in acetonitrile, 80:20, v/v Sample temp.: 25°C
Manufacturer C
Columns: Dimensions: Particle size: Pore size: Conditions: Mobile phase:
Manufacturer D Flow: Temp.: Det.:
Wide Pore C18 150mm x 2.1mm 5µm 300Å A: 0.10% TFA in water, B: 0.10% TFA in acetonitrile, 20% B to 70% B in 30 min. 0.20mL/min. 25ºC (or ambient) UV @ 214nm
LC_0324
PEGylated Oxytocin on Viva C18 (LC/MS)
free literature Viva Wide Pore HPLC Columns Exceptional Performance for Peptides, Proteins, or Other Large Molecules Download your free copy from www.restek.com.
LC_PH0467
Sample: Inj.: Conc.: Sample diluent:
oxytocin PEGylation reaction products 20µL 300pmoles/µL 0.1% formic acid in water (v:v)
Column: Cat.#: Dimensions: Particle size: Pore size:
Viva C18 9514561 150mm x 1.0mm 5µm 300Å
Conditions: Mobile phase: Flow: Temp.: Det.: Interface: Ion mode: Temp.: Capillary: Cone:
0.1% formic acid in water:0.1%formic acid in acetonitrile, 60:40 0.1mL/min. ambient Micromass Quattro II ESI positive 200°C 2.25kV 40V
Flyer lit. cat.# 59939
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HPLC CHROMATOGRAMS | CLINICAL/FORENSIC
Pain Management Drugs Pain Management Drugs in Urine on Ultra II® Biphenyl (LC/MS/MS)
NEW! Peaks RT (min.) 1. Acetaminophen 1.18 2. Morphine 1.79 3. Codeine 3.21 4. Oxycodone 3.46 5. Hydrocodone 3.60 6. Fentanyl 6.61 7. Buprenorphine 6.66 8. Lorazepam 6.79 9. Diazepam 8.06 10. Methadone 8.21
MRM1 152/110 286/157 300/165 316/298 300/199 337/132 468/187 321/275 285/154 310/77
MRM2 152/65 286/181 300/215 316/241 300/171 337/216 468/84 321/229 285/193 310/223
Recommended for opiates.
LC_CF0516
Column Dimensions: Particle Size: Pore Size: Temp.: Sample Conc.: Inj. Vol.: Mobile Phase
Detector Model #: Ion Source: Ion Mode: Curtain Gas: Gas 1: Gas 2: Source Temp.: Source Voltage: Mode: Dwell Time: Instrument
Ultra II® Biphenyl (cat.# 9609352) 50 mm x 2.1 mm ID 3 µm 100 Å 40 °C 5 ng/mL in 1:10 urine:mobile phase 5 µL A: 0.1% formic acid in water B: 0.1% formic acid in methanol Time (min.) Flow (mL/min.) 0 0.5 10 0.5 10.1 0.5 12 0.5
%A 90 0 90 90
%B 10 100 10 10
Acknowledgement
Applied Biosystems/MDS Sciex LC/MS/MS API-5000™ TurboIonSpray® ESI+ 25 psi (172.4 kPa) 60 psi (413.7 kPa) 40 psi (275.8 kPa) 550 °C 2000 V MRM 50 ms Applied Biosystems/MDS Sciex LC/MS/MS System Special thanks to Applied Biosystems for providing instrument time.
For more information on Biphenyl columns, visit
www.restek/biphenyl www.restek.com Website : www.chromtech.net.au E-mail :
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Page 748
HPLC CHROMATOGRAMS | CLINICAL/FORENSIC
Opiates, THC Opiates on Allure® PFP Propyl (LC/MS/MS)
Conditions: Instrument: Mobile phase:
1. morphine 2. oxymorphone 3. hydromorphone 4. codeine 5. 6-monoacetylmorphine 6. oxycodone 7. hydrocodone
Shimadzu Prominence HPLC A: 0.1% formic acid in water B: 0.1% formic acid in 80:20, methanol:acetonitrile Time: 0.0 3.00 6.00 6.10 8.10
%B 10 50 50 10 Stop pumps
Sample: Inj.: Conc.: Solvent:
opiates 10µL 25ug/mL methanol
Flow: Temp.:
0.40mL/min. 30°C
Column: Cat.#: Dimensions: Particle size: Pore size:
Allure® PFP Propyl 9169552 50mm x 2.1mm 5µm 60Å
Det.: Ion source: IonSpray voltage: Gas 1: Gas 2: Source temp.:
Applied Biosystems/MDS Sciex API 3200™ MS/MS system Electrospray, positive 5,500 65psi (448kPa) 45psi (310kPa) 600°C
Compound morphine morphine hydromorphone hydromorphone oxymorphone oxymorphone codeine codeine hydrocodone hydrocodone oxycodone oxycodone 6-monoacetylmorphine 6-monoacetylmorphine
LC_PH0454
Q1 286 286 286 286 302 302 300 300 300 300 316 316 328 328
Mass Spectrometer Experiments: Declustering Collision Q3 Potential (V) Energy (V) 152 46 79 165 46 51 185 46 41 157 46 55 227 36 37 198 36 55 152 46 85 115 46 89 199 46 39 128 46 69 240 31 39 256 31 33 211 51 55 193 51 35
THC and Metabolites on Ultra II® Biphenyl (LC/MS/MS)
NEW!
Peaks 1. Cannabidiol 2. THC-COOH-gluc 3. THC-COOH 4. Cannabinol 5. Δ9-THC
MRM1 (Q1/Q3) 315/123 345/299 345/299 311/223 315/193
Column Dimensions: Particle Size: Pore Size: Temp.:
Ultra II® Biphenyl (cat.# 9609853) 50 mm x 3.0 mm ID 2.2 µm 100 Å 35 °C
Sample Diluent: Conc.: Inj. Vol.: Mobile Phase
Acetonitrile 50 ng/mL 20 µL A: 0.1% formic acid in water B: 0.1% formic acid in methanol
MRM2 (Q1/Q3) 315/193 345/193 345/193 311/178 315/123
Time (min.) 0 0.50 3.50 4.50 4.60 6.00
%B 50 50 100 100 50 50
Flow:
0.7 mL/min.
Detector Model #: Ion Source: Ion Mode: Ion Spray Voltage: Curtain Gas: Gas 1: Gas 2: Source Temp.: Mode: Dwell Time: Instrument
Applied Biosystems/MDS Sciex LC/MS/MS 3200 Q-Trap TurboIonSpray® ESI+ 5 kV 25 psi (172.4 kPa) 50 psi (344.7 kPa) 60 psi (413.7 kPa) 550 °C MRM 50 ms Applied Biosystems/MDS Sciex LC/MS/MS System
LC_CF0512
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HPLC CHROMATOGRAMS | CLINICAL/FORENSIC
Alcohol Metabolites, Diuretics Alcohol Metabolites on Ultra II® Biphenyl (LC/MS)
NEW!
Sample: Inj.: 5µL Conc.: 5µg/mL each component Sample diluent: urine diluted 1:10 with mobile phase
Peak List 1. matrix 2. ethyl glucuronide 3. ethyl sulfate
m/z 125 221 125
k' — 2.75 4.02
Column: Cat.#: Dimensions: Particle size: Pore size:
Intensity — 220,408 236,693
Conditions: Instrument: Mobile phase:
Flow: Temp.: Det.: Ion source: Mode: Source temp.: Source voltage:
Ultra II® Biphenyl 9609552 50mm x 2.1mm 5µm 100Å Shimadzu Prominence UFLCXR A: 5mM dihexylammonium acetate in water B: 5mM dihexylammonium acetate in methanol Time (min.) %B 0.0 40 2.5 40 2.6 100 5.4 100 5.5 40 7.5 40 0.3mL/min. 40°C Shimadzu LCMS-2010EV ESISIM 200°C -4,000V
LC_CF0500
Diuretics on Ultra II® Biphenyl (LC/MS/MS, Negative Run) MRM1 (Q1/Q3) 1. Acetazolamide 221/83 2. Chlorothiazide 294/214 3. Hydrochlorothiazide 296/78 4. Bendroflumethiazide 420/289 5. Furosemide 329/285
Diuretics in Urine on Ultra II® Biphenyl (LC/MS/MS, Positive Run) MRM2 (Q1/Q3) 221/80 294/179 296/269 420/328 329/205
MRM1 (Q1/Q3) 1. Amiloride 230/171 2. Triamterene 254/237 3. Indapamine 366/132 4. Ethacrynic acid 303/179 5. Bumetanide 365/240
MRM3 (Q1/Q3) 221/58 294/215 296/205 420/197 329/78
MRM2 (Q1/Q3) 230/116 254/104 366/91 303/194 365/156
MRM3 (Q1/Q3) 230/60 254/141 366/117 303/257 365/284
NEW!
Good retention for hydrophilic compounds! Ion Mode: ESI+ Ion Spray Voltage: 5.5 kV
Ion Mode: ESIIon Spray Voltage: -4.5 kV
LC_CF0510
Column Dimensions: Particle Size: Pore Size: Temp.:
Ultra II Biphenyl (cat.# 9609352) 50 mm x 2.1 mm ID 3 µm 100 Å 40°C
Sample Conc.: mobile phase Inj. Vol.:
500 ng/mL diuretics in urine, diluted 10x in
®
LC_CF0509
Mobile Phase A: 0.1% formic acid in water B: 0.1% formic acid in methanol Time (min.) Flow (mL/min.) 0 0.5 6.00 0.5 6.1 0.5 8.00 0.5
%A 90 0 90 90
%B 10 100 10 10
Detector Model #: Ion Source: Curtain Gas: Gas 1: Gas 2: Source Temp.: Mode: Dwell Time:
Applied Biosystems/MDS Sciex LC/MS/MS API 5000 TurboIonSpray® 25 psi (172.4 kPa) 60 psi (413.7 kPa) 40 psi (275.8 kPa) 550°C MRM 50 ms
5 µL
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HPLC CHROMATOGRAMS | CLINICAL/FORENSIC
Amphetamines, Cocaine Amphetamines on Ultra II® Biphenyl (LC/MS)
NEW! Peaks 1. Amphetamine 2. Methamphetamine 3. MDA 4. MDMA 5. MDEA
RT (min.) 2.812 3.432 3.537 4.092 4.679
Column Dimensions: Particle Size: Pore Size: Temp.: Sample Diluent: Conc.: Inj. Vol.: Mobile Phase
m/z 135.95 149.95 179.95 193.95 207.95
Ultra II® Biphenyl (cat.# 9609552) 50 mm x 2.1 mm ID 5 µm 100 Å 30 °C mobile phase 2 µg/mL 5 µL A: 0.1% formic acid in water B: 0.1% formic acid in methanol Time (min.) Flow (mL/min.) 0 0.3 10 0.3 10.1 0.3 12.0 0.3
%A 80 5 80 80
%B 20 95 20 20
Detector Shimadzu 2010EV Acquisition Type: Scan Scan Speed: 2000 amu/sec. Heat Block: 200 °C Interface: ESI+ Interface Temp.: 250 °C Scan Range: 100 - 250 amu Event Time: 0.1 sec. Instrument Shimadzu UFLCXR Notes Data were collected in scan mode. An extracted ion chromatogram is shown. LC_CF0518
For more information on Biphenyl columns, visit
www.restek/biphenyl
Cocaine and Ecgonine Methyl Ester on Allure® PFP Propyl (LC/MS/MS) Conditions: Mobile phase:
Peak List: 1. EME (ecgonine methyl ester) 2. COC (cocaine)
Flow: Column temp.: Det.:
Sample: Inj.: Conc.: Solvent: Temp.:
10µL 1µg/mL water 4°C
Column: Cat.#: Dimensions: Particle size: Pore size:
Allure® PFP Propyl 9169532 30mm x 2.1mm 5µm 60Å
Interface: Interface temp.: Ion mode: ESI probe voltage: Orifice: Ring: Collision gas: Collision gas pressure: Collision gas energy: Electron multiplier: Auxilary gas flow: Nebulizer gas setting: Curtain gas setting:
1
XIC of +MRM (3 pairs) for 200.0 / 182.0 amu
0
0.5
1.0
1.5
5mM ammonium formate, pH 3.0: acetonitrile (10:90, v/v) 0.6mL/min. ambient Applied Biosystems/MDS SCIEX API 3200™ MS/MS system turbo ion spray, ESI 150°C positive 5,000V + 71V + 265V nitrogen 2.2 mTorr 28 eV (COC) 26 eV (EME) 2,100 volts 7,000cc/min. 15lb/in.2 12lb/in.2
Get More! Clinical, Forensic & Toxicology Solutions Online www.restek.com/CFT
1.30e3 cps
2.0
2.5
3.0
3.5
4.0
4.5
min.
2
XIC of +MRM (3 pairs) for 304.0 / 182.0 amu
0 0.5 1.0 1.5 2.0 2.5 Data Courtesy of Shane Needham, Pfizer Inc.
2.35e3 cps
3.0
3.5
4.0
4.5
min.
LC_0126
750 www.restek.com Website : www.chromtech.net.au E-mail :
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HPLC CHROMATOGRAMS | CLINICAL/FORENSIC
Benzodiazepines Benzodiazepines on Allure® PFP Propyl (LC/MS/MS)
Sample: Inj.: Conc.: Solvent:
benzodiazepines 20µL NA NA
Column: Cat.#: Dimensions: Particle size: Pore size:
Allure® PFP Propyl 9169552 50mm x 2.1mm 5µm 60Å
Analyze 27 benzodiazepines in less than 10 minutes!
Conditions: Instrument: Shimadzu Prominence HPLC Mobile phase: A: 0.1% formic acid and 1mM ammonium formate in water B: 0.1% formic acid and 1mM ammonium formate in acetonitrile Time (min.) 0.0 10.00 15.00 15.50 17.50 Flow: Temp.: Det.: Ion Source: IonSpray Voltage: Gas 1: Gas 2: Source Temp.:
Flow (µL/min.) 500 1000 1000 500 500
%B 10 90 90 10 10
see gradient table 40°C Applied Biosystems/MDS Sciex API 3200™ MS/MS system Electrospray, positive NA NA NA 500°C
Data courtesy of: Applied Biosystems MDS Sciex
LC_PH0463
MRM Transitions, retention times, and LOQ values.
Bar color indicates shared precursor ions. Note compounds with shared precursor ions are baseline resolved on the Allure® PFP Propyl column, as shown by retention time comparison. Data courtesy of Applied Biosystems MDS Sciex.
Compound Name
Retention Time (min.) 7-aminonitrazepam 3.2 7-aminoclonazepam 3.3 7-aminoflunitrazepam 3.8 Bromazepam 3.8 α-hydroxyalprazolam 4.1 α-hydroxytriazolam 4.1 Oxazepam 4.2 Lorazepam 4.3 Estazolam 4.4 Zaleplon 4.4 2-hydroxyethylflurazepam 4.5 Desmethylflunitrazepam 4.5 Nitrazepam 4.6 Clonazepam 4.7 Desalkylflurazepam 4.7 Temazepam 4.7 Triazolam 4.7 Alprazolam 4.8 Lormetazepam 4.8 Clobazam 4.9 Flunitrazepam 5.0 Nordiazepam 5.0 Zolpiclone 5.4 D5-Diazepam 5.4 Diazepam 5.5 Chlordiazepoxide 6.0 Prazepam 6.1 Zolpidem 7.4 Midazolam 7.9 Flurazepam 8.5 Medazepam 9.0 D3-Doxepine 9.1
Precursor Ion (amu) 252.1 286.1 284.1 316.0/318.0 325.1 359.0 287.0 321.0/323.1 295.0 306.2 333.1 300.1 282.0 316.0 289.1 301.1/303.1 343.0 309.1 335.0/337.1 301.1 314.0 271.1 389.1 290.1 285.0 300.1 325.1 308.1 326.1 388.2 271.0 283.0
MRM 1 (amu) 121.1 121.0 135.1 182.1 297.2 239.2 241.1 275.0 205.0 236.3 211.2 254.2 236.1 270.2 140.1 255.1 238.9 205.1 289.0 259.1 268.1 140.2 244.8 198.2 193.2 227.1 271.1 235.1 291.3 315.1 91.1 107.1
MRM 2 (amu) 94.0 222.2 226.0 182.1 204.9 176.0 268.9 277.0 267.1 264.2 109.0 198.2 180.2 214.0 226.1 257.2 314.9 281.1 291.1 224.3 239.1 164.9 217.0 154.1 283.2 140.0 236.1 222.0 317.1 207.3 -
DP 51 46 51 51 51 61 41 41 51 56 56 56 71 56 71 35 61 56 41 46 56 46 16 55 55 36 81 56 56 36 46 41
CE (MRM 1) 35 41 39 45 31 63 27 31 53 35 51 35 35 41 41 30 53 53 29 29 35 37 25 41 41 31 31 39 33 27 41 35
CE LOQ (MRM 2) (ng/mL) 53 1.0 35 0.5 49 0.5 45 5.0 59 2.0 37 5.0 19 10.0 27 5.0 31 2.0 27 0.5 41 1.0 51 2.0 51 2.0 51 2.0 39 2.0 30 5.0 37 1.0 35 1.0 29 2.0 47 1.0 49 1.0 35 2.0 41 1.0 37 1.0 21 5.0 53 2.0 35 0.2 63 0.5 27 0.1 39 2.0 -
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