LC-MS ON HPLC COLUMNS

Transcription

1 LC-MS PLC CLUMS

2 o.ti9e LC-ESI-MS application 75 x 2 mm 2 1 nilvadipine 1 2 nisoldipine min Cadenza CD-C18, 75 x 2. mm 6 acetonitrile,.2 ml/min, 4 C egative-esi TI9E-ZJ6

3 o.ti54e LC-MS/MS igh Throughput Analysis LC beta-estradiol 1 pg/ul, 1uL injection min [LC] Cadenza CD-C18, 5 x 2 mm methanol / water = 75 / 25.2 ml/min, 4 C [MS] API 4 ESI-negative MS 3 C m/z 271 beta-estradiol MS/MS Courtesy of TAKARA SUZ C., LTD. TI54E-AI19

4 o.ti55e LC and FAB Mass Analyses of D-Amino Acid-containing Peptides LC LC FAB-MS FAB-MS [LC] Cadenza CD-C18, 75 x 4.6 mm 5-45 of methanol/acetonitrile (6:4) in 5 mm phosphate, p ml/min Courtesy of A. Sugihara, saka Municipal Technical Research Institute TI55E-AI27

5 o.ti76e LC-MS Application of Myoglobin Tryptic Digest mau UV at 215 nm Ints [MS] Esquire 3plus ( Ion Trap MS, positive mode ) BPC MS Time [min] Cadenza CD-C18, 15 x 2 mm A:.1 TFA in water B:.1 TFA in acetonitrile 2-5 B ( - 5 min ).2 ml/min, 45 C Myoglobin (orse eart), Tryptic digest Courtesy of D.igo, Bruker Daltonics K.K. TI76E-BI1

6 o.ti85e igh Throughput LC-MS Application for Local Anesthetic and Antiarrhythmic Drugs C3 C 2 CC 2 C C3 1 atenolol Int. 16e3 15e3 14e3 13e3 12e3 11e3 1e3 9e3 8e3 7e3 Shimadzu LCMS-21A ( ESI, positive, SIM ) C2 C 2 C 2 C2C2 C2C3 C2C3 C3 C 2 CC 2 C C3 C3 C 2 CC 2 C C3 procaine pindolol metoprolol 6e3 C2C2C3 5e3 4e3 3e3 5 C3 C 2 CC 2 C C3 propranolol 2e3 1e min 6 C3 C 2 CC 2 C C3 C 2 C C2 alprenolol 7 C C2C2 C3 C3 tetracaine (C2)3C3 Cadenza CD-C18, 3 x 2 mm A: 1mM ammonium acetate B: acetonitrile - 9 B ( - 1 min), 9 B (1-2 min).5 ml/min, 4 C 1 pg/ul, 5uL C3 C3 C C3 C3 C C2C3 C2 C2C3 C2C2 C2C3 C2C3 (C2)3C3 C C2(C2)2C3 lidocaine dibucaine bupivacaine TI85E-BJ2

7 o.ti91e LC-MS Application for Endocrine Disruptor API4 (TAKARA BI IC. ) C octylphenol (Mw ) 5 pg min Cadenza CD-C18, 1 x 2 mm 8 methanol,.2 ml/min, 4 C, 5µL(1pg/µL) API4: ESI, MRM egative, Q1/Q3 : 25 / 16 Courtesy of J.Watanabe, TAKARA BI IC. TI91E-CF4

8 o.ti92e LC-MS Application for Substances in Ginkgo Leaves API4 (TAKARA BI IC. ) 3C R2 ginkgolid A Mw ginkgolid B Mw ginkgolid C Mw R1 R2 R1 C3 C3 C3 C A B 5 fg min Cadenza CD-C18, 1 x 2 mm 6 methanol,.2 ml/min, 4 C, 5µL(1fg/µL) API4: ESI, MRM egative Q1/Q3: ginkgolid A 47/351 ginkgolid B 423/367 ginkgolid C 439/383 Courtesy of J.Watanabe, TAKARA BI IC. TI92E-CF4

9 o.ti93e LC-MS Application for Bug Repellent Active Ingredient API3 (TAKARA BI IC. ) 3C 3C C3 DEET (,-diethyl-m-toluamide) Mw fg min Cadenza CD-C18, 5 x 2 mm.1 formic acid / 6 methanol,.2 ml/min, 4 C, 5µL (2fg/µL) API3: ESI, MRM Positive Q1/Q3: 192/119 Courtesy of J.Watanabe, TAKARA BI IC. TI93E-CF4

10 o.ti94e LC-MS Application for Capsaicins API4 (TAKARA BI IC. ) C 3 C 3 1 pg Me capsaicin Mw. 35 Me C 3 C dihydrocapsaicin Mw min Cadenza CD-C18, 1 x 2 mm.1 acetic acid / 75 methanol,.2 ml/min, 4 C, 5µL (2fg/µL) API4: ESI, MRM Positive Q1/Q3: capsaicin 36/137 dihydrocapsaicin 38/137 Courtesy of J.Watanabe, TAKARA BI IC. TI94E-CF5

11 o.ti135e Volicitin-related Compounds from the ral Secretion of Caterpillars 1 3 MS UV R 3 4 R' R =, R =, 1 3 C C2 5 6 R' =, 2 R' =, 4 C C2 R Cadenza CD-C18, 75 x 3 mm A:.5 acetic acid in water B:.8 acetic acid in acetonitrile 2-95B ( -4min ).2 ml/min, 6 C, 1 µl egative ESI mass, UV at 2 nm R =, 5 R =, 7 6 Courtesy of Dr. aoki Mori, Kyoto Univ. TI135E-CG25

12 o.ti189e - Solanine and - Chaconine from Potato -D-Gal -D-Glu -D-Glu -L-Rha -L-Rha -L-Rha - solanine - chaconine Ion Intensity (cpm) PLC-ESI-MS m/z m/z solanine - chaconine min Cadenza CD-C18, 75 x 2 mm.1 trifluoroacetic acid / acetonitrile = 75 / 35.2 ml/min, 35 deg.c, LCMS-21A, ESI positive Courtesy of Dr. Fumio MATSUDA and Prof. isashi MIYAGAWA, Department of Agriculture, Kyoto University. TI189E-D25

13 o.ti25e LC-MS Application for Antiseptic Substance Cl Cl chlorhexidine Mw API4 (TAKARA BI IC. ) pg Time, min Max cps. 1.4e5 1.4e5 1.3e5 1.2e5 Chlorhexidine Results.rdb (C): "Linear" Regression ("1 / x" weighting): y = 1.43e+4 x (r =.9999) 1.1e5 1.e5 9.e4 r = e4 7.e4 6.e4 5.e4 4.e4 3.e4 2.e4 1.e Concentration, ng/ml Unison UK-C8, 5 x 2 mm.1 formic acid / acetonitrile = 2 / 1,.2 ml/min, 5uL(5 fg/ul) API4: ESI, MRM Positive, Q1/Q3 : 55 / 336 Courtesy of J.Watanabe, TAKARA BI IC. TI25E-DK3

14 o.ti26e LC-MS Application for Calcium Antagonist API4 (TAKARA BI IC. ) 1.35 Max cps fg S C3 C 3 C 3 Cl 2 1 C 3 diltiazem Cl Mw Time, min Results.rdb (Diltiazem): "Linear" Regression ("1 / x" weighting): y = 5.98e+5 x (r =.9999) 3.e6 2.8e6 2.6e6 2.4e6 2.2e6 2.e6 1.8e6 r = e6 1.4e6 1.2e6 1.e6 8.e5 6.e5 4.e5 2.e Concentration, ng/ml Unison UK-C8, 5 x 2 mm.1 formic acid / acetonitrile = 2 / 1,.25 ml/min, 5 ul(5 fg/ul) API4: ESI, MRM Positive, Q1/Q3 : 415 / 178 Courtesy of J.Watanabe, TAKARA BI IC. TI26E-DK3

15 o.ti27e LC-MS Application for Coenzyme Q1 API4 (TAKARA BI IC. ) Max cps. C 3 12 Me fg Me C Coenzyme Q1 Mw Time, min CoenzymeQ1 Result.rdb (CoenzymeQ1): "Linear" Regression ("1 / x" weighting): y = 6.66e+3 x (r =.9998) 1.3e6 1.2e6 1.1e6 1.e6 9.e5 8.e5 r = e5 6.e5 5.e5 4.e5 3.e5 2.e5 1.e Concentration, ng/ml Unison UK-C8, 75 x 3 mm.1 formic acid / acetonitrile / isopropanol = 1 / 5 / 5.8 ml/min, 1 ul(5 fg/ul) API4: APCI, MRM Positive, Q1/Q3 : / 197 Courtesy of J.Watanabe, TAKARA BI IC. TI27E-DL1

16 o.ti312e LC-MS/MS Application of Water Soluble Vitamins ist-1x-2 1 ist-1x-2 1 ist-1x-2 1 ist-1x : MRM of 2 Channels ES > e : MRM of 2 Channels ES > e5 5: MRM of 2 Channels ES > e3 5: MRM of 2 Channels ES+ 22 > 9 5.6e5 ist-1x : MRM of 2 Channels ES+ 123 > 8 1.8e6 ist-1x : MRM of 2 Channels ES+ 17 > e6 ist-1x : MRM of 1 Channel ES+ 177 > e5 ist-1x : MRM of 1 Channel ES+ TIC 1.77e Time Unison UK-C18, 25 x 2 mm A:.1 C in water B:.1 C in acetonitrile B (-5min), -5B (5-15min), 5-95B (15-17min), B (17-25min).25mL/min, ambient, 5uL ESI-MS, MRM Positive Courtesy of Dr. Pei Chen, USDA, ARS, BRC, Food Composition Lab - Beltsville, MD, USA TI312E-GD13

17 o.ti338e LC-MS/MS Application of Marajuana and it s Metabolite in Blood TC delta-9tetrahydrocannabinol TC-C Unison US-C18, 5 x 2 mm A:.1 C in water, B:.1 C in acetonitrile 5-9B in 5 min,.5 ml/min, 28 deg. C 32 Q Trap -2.3 min, ESI egative, min, ESI Positive Whole blood, SPE extraction LD TC =.1ng/mL LD TC-C =.5ng/mL Courtesy of Massachusetts State Crime Lab., USA TI338E-GG7

18 o.ti365e LC-MS/MS Application of piates in Whole Blood : morphine 2: oxymorphone 3: hydromorphone 4: codeine 5: oxycodone 6: 6-acetyl morphine 7: hydrocodone 4 1 Unison US-C18, 5 x 2 mm A:.1 C in Water B:.1 C in Acetonitrile 5B (-.5min) 5-25B (.5-2.mm) 25-9B ( min) 9B ( min).6 ml/min, 28 deg. C 32 Q-Trap, ESI Positive Whole blood, SPE Extraction LD =.5ng/mL; LQ = 1.ng/mL Courtesy of Massachusetts State Crime Lab., USA TI365E-GJ9

19 o.ti396e LC-MS/MS Application - rganophosphorous Pesticides methamidophos 3C 2 P S C3.5 ppb Unison UK-C18, 5 x 2 mm Mobile phase:.1 C in 5 Me flow rate:.25 ml/min Column temp.: 4 deg.c Ionization: ESI positive SRM transition: m/z 142 -> m/z 94, CID 16V dichlorvos P 3C C C Cl Cl C3 1 ppb Unison UK-C18, 5 x 2 mm Mobile phase:.1 C in 4 Me flow rate:.25 ml/min Column temp.: 4 deg.c Ionization: ESI positive SRM transition: m/z 221 -> m/z 19, CID 18V Courtesy of Dr. T. ayama, Environmental Science Center, Foundation for Kyushu Environmental and ccupational ealth, Japan TI396E-B27

20 o.ti397e LC-MS/MS Application - Steroid ormones and Metabolites Intensity, cps Intensity, cps 1.5e4 1.4e4 1.3e4 1.2e4 1.1e4 1.e e alphahydroxyprogesterone Q1/Q3 = 331/97 25 pg testosterone Q1/Q3 = 289/97 1 pg Time, min C3 C estriol Q1/Q3 = 287/171 1 pg Time, min 5alpha-dihydrotestosterone Q1/Q3 = 291/255 3 pg progesterone Q1/Q3 = 315/19 1 pg Unison UK-C8, 5 x 2 mm.1 C aq. ->.1 C in AC.2 ml/min, 1uL API4 (TAKARA BI IC.) ESI, MRM positive C3 C estrone Q1/Q3 = 269/145.1 pg 17beta-estradiol Q1/Q3 = 271/145 1 pg Unison UK-C8, 5 x 2 mm 1mM Ac4 -> acetonitrile.2 ml/min, 1uL API4 (TAKARA BI IC.) ESI, MRM negative Intensity, cps 1.15e4 1.1e4 1.e C3 C pregnanetriol Q1/Q3 = 354[M+4]+ / 31 3 fg C Time, min C3 pregnanediol Q1/Q3 = 338[M+4]+ / fg Unison UK-C8, 5 x 2 mm 1mM Ac4 -> methanol.2 ml/min, 1uL API4 (TAKARA BI IC.) ESI, MRM positive Courtesy of TAKARA BI IC. TI397E-C7

21 o.ti411e LC-MS/MS Application -Tetracycline Antibiotics 2859_TC_ MRM of 6 Channels ES+ TIC 3.4e _TC_2 MRM of 6 Channels ES > e _TC_2 MRM of 6 Channels ES > e _TC_2 MRM of 6 Channels ES > e3 CTC first ion _TC_2 MRM of 6 Channels ES > e3 CTC second ion TC first ion TC second ion Cl oxytetracycline (TC) chlortetracycline (CTC) _TC_2 MRM of 6 Channels ES > 41.1 TC first ion 1.74e _TC_2 MRM of 6 Channels ES > e4 TC second ion Time tetracycline (TC) 2 Cadenza CW-C18, 1 x 2 mm + Guard column A:.1 C aq., B:.1 C in AC 5-95B ( - 7 min ), 95-5B ( 7-8 min ), 5B ( 8-17 min ).2 ml/min, 4 deg.c, 1 ul ( 2 ppb ), ESI, positive Courtesy of Mr.. MAEDA, ASSCIATI F MEAT SCIECE & TECLGY ISTITUTE, JAPA TI411E-F24

22 o.ti438e LC-MS/MS: Melamine and Related Compounds melamine 2. ammeline 3. cyanuric acid 4. ammelide Positive 1 1 ppb egative 4 3 ppb 2 6 ppb 3 2 ppb Unison UK-Amino, 15 x 3 mm A: acetonitrile, B: 1mM ammonium acetate +.1 acetic acid 25B (Isocratic).4 ml/min (5.9MPa), 4 deg.c, 5uL TI438E-J3

23 o.ti44e A proteomics study on human breast cancer cell lines by fluorogenic derivatization-liquid chromatography/tandem mass spectrometry DAABD-proteins Breast cancer cell _ 1 Breast cancer cell _ 2 Breast cancer cell _ 3 Breast cancer cell _ 4 Breast cancer cell _ 5 Breast cancer cell _ 6 Breast cancer cell _ 7 ormal mammary epitherial cell Ref-1: Biomedical Chromatography, 28, 22 (11), Ref-2: Journal of Proteome Research, 27, 6, Intrada WP-RP, 25 x 4.6 mm A:.15 TFA in (acetonitrile /isopropanol /water = 9 /1 /9) B:.15 TFA in (acetonitrile /isopropanol /water = 69 /1 /3) C:.2 TFA in (acetonitrile /isopropanol /water = 9 /1 /9) 5B,1C(-5min), 5-3B,1-35C (5-3min), 3B,35C(3-65min) 3-35B,35C(65-7min), 35-38B,35C(7-13min) 38-44B,35-55C(13-25min), 44B,55C(25-3min) 44-47B,55-53B(3-33min), 47-6B,53-4C(33-48min) 6-7B,4-3C(48-52min), 7-9B,3-1C(52-57min).55 ml/min, 6 deg.c, FLD( Ex.395nm, Em.55nm) Courtesy of Kazuhiro Imai, Ph.D. Dhc., Professor & Dean, Musashino University, Japan TI44E-J15

24 o.ti463e LC-MS/MS Application for Duloxetine and Ethyl Morphine in Whole Blood XIC of +MRM (4 pairs): 298.2/44.1 amu from Sample 3 (1.) of 19.wiff (Turbo Spray) Max cps. I... I... I... I Time, min XIC of +MRM (4 pairs): 298.2/44.1 amu from Sample 3 (1.) of 19.wiff (Turbo Spray) Max cps Time, min XIC of +MRM (4 pairs): 298.2/154.3 amu from Sample 3 (1.) of 19.wiff (Turbo Spray) Max cps Time, min XIC of +MRM (4 pairs): 314.2/152.2 amu from Sample 3 (1.) of 19.wiff (Turbo Spray) Max cps Ethyl morphine Time, min XIC of +MRM (4 pairs): 314.2/128.3 amu from Sample 3 (1.) of 19.wiff (Turbo Spray) Max cps. TIC Duloxetine Duloxetine I Ethyl morphine Time, min Unison US-C18, 5 x 2 mm A:.1 C in water, B:.1 C in acetonitrile 5B (-.5min), 5-9B (.5-4.5min), 5B (4.5-6min).5mL/min (LD.5 ng/ml, LQ.1 ng/ml) Whole Blood, SPE Extraction 32 Q-Trap: ESI, MRM Positive Duloxetine: m/z > 44.1 and Ethyl Morphine (I.S.): m/z > and Ref) Rapid Quantification of Duloxetine in Blood and Urine by LC-ESI-MS/MS Albert Elian, Massachussetts State Police Crime Lab, USA Society of Forensic Toxicologists (SFT), 28, USA Courtesy of Massachusetts State Police Crime Lab., USA TI463E-L1

25 o.ti464e LC-MS/MS Application for 25-ydroxy Vitamin D3 XIC of +MRM (1 pair): 383.3/211.1 amu from Sample 5 (fresh std 5ng/ml opt conditions_inj 2uL) of Vit D3 stds_imtakt e Max. 5.6e4 cps. 5.e4 4.5e4 3C 3C C3 C3 4.e4 3.5e4 3.e4 C2 2.5e4 2.e4 25-ydroxy Vitamin D3 (Calcifediol) 1.5e4 1.e Time, min Cadenza CD-C18, 15 x 4.6 mm A: water / methanol = 75 / 25 B: acetonitrile / ethyl acetate = 7 / 3 4B (-1min), 4-95B (1-8min), 95B (8-15min), 4B (15-22min).7mL/min, ambient Applied Biosystems 4 Q trap Ionization: APCI Positive mode LQ: 1ng/mL Courtesy of Penn State ershey Medical Center, USA TI464E-L1

26 o.ti477e LC-MS application for phospholipids :/2:4-PI :/18:1-PC 2 1: TF MS ES e3 1: TF MS ES e4 C2CR1 R2 CC C2 P Phosphatidylinositol (PI) :/18:2-PC 3 1: TF MS ES e3 C2CR1 R2 CC C2 P Phosphatidylcholine (PC) C2C2(C3) Fused Core TM 1: TF MS ES e4 1: TF MS ES e4 1: TF MS ES- TIC 1.4e a 4.19 b :1/18:1-PE (2.7um), DS, 1 x 2.1 mm Time 1: TF MS ES TIC TIC, 5 x 1 4 FS 5.5e a 16:/18:1-PG b d18:1/c16:-sm 5 6 TIC, 1 x 1 5 FS C2CR1 R2 CC C2 P Phosphatidylethanolamine (PE) C2C23 C2CR1 R2 CC C2 P C2CC2 Phosphatidylglycerol (PG) C R1 R2C C C2 P C2C2(C3)3 Sphingomyelin (SM) Scan Cadenza CW-C18, 1 x 2 mm 5mM C4 in (acetonitrile / methanol = 1 / 1).2mL/min, 2deg.C ESI-TFMS, egative Courtesy of Yasuko Yoshioka, Inst. ealth Biosciences, Univ. of Tokushima, Japan TI477E-IA8

27 o.ti487e LC-MS/MS Application for Sphingolipids (Ceramide) XIC of +EMS: to amu from Sample 1 (C16 Ceramide) of March_11_9_p... Max. 3.3e8 cps. 3.3e8 3.e8 2.5e8 16: Ceramide (C16 Ceramide) -Palmitoylsphingosine e8 1.5e8 +EMS: to min from Sample 1 (C16 Ceramide) of March_11_9_pos_EM e6 9.e6 8.e6 7.e6 6.e Max. 9.6e6 cps. 5.e e8 5.e7. 4.e6 3.e6 2.e6 1.e m/z, amu Time, min Cadenza CW-C18, 15 x 2 mm A: [water / C (1:.2 v/v)] containing 7mM ammonium formate B: [methanol / C (1:.2 v/v)] containing 5mM ammonium formate 7-9B (-1min), 9B (1-1.5min), 9-1B ( min), 1B (2.5-6min), 1-4B (6-6.5min), 4B (6.5-1min).2mL/min -.28mL/min (-6min),.2mL/min (6-1min) 48deg.C, 5uL (5ug / ml) Applied Biosystems QTrap, ESI Positive [M-2+] Courtesy of Jennifer Ro and Joe Williams Department of Evolution, Ecology, and rganismal Biology, hio State University - USA TI487E-IC2

28 o.ti488e LC-MS/MS Application for Sphingolipids (Sphinganine) XIC of +EMS: 32. to 34.9 amu from Sample 1 (sphinganine) of March_11_9_pos... Max. 2.3e8 cps. 2.3e8 2.2e8 2.e Sphinganine (Dihydrosphingosine) 2 1.8e8 1.6e8 1.4e8 1.2e8 1.e8 8.e7 +EMS: to min from Sample 1 (sphinganine) of March_11_9_pos_EMS_ e6 3.e6 2.5e6 2.e6 1.5e6 6.e7 1.e e7 2.e e m/z, amu Time, min Cadenza CW-C18, 15 x 2 mm A: [water / C (1:.2 v/v)] containing 7mM ammonium formate B: [methanol / C (1:.2 v/v)] containing 5mM ammonium formate 7-9B (-1min), 9B (1-1.5min), 9-1B ( min), 1B (2.5-6min), 1-4B (6-6.5min), 4B (6.5-1min).2mL/min -.28mL/min (-6min),.2mL/min (6-1min) 48deg.C, 5uL (5ug / ml) Applied Biosystems QTrap, ESI Positive Courtesy of Jennifer Ro and Joe Williams Department of Evolution, Ecology, and rganismal Biology, hio State University - USA TI488E-IC2

29 o.ti489e LC-MS/MS Application for Sphingolipids (Sphingosine) XIC of +EMS: to amu from Sample 1 (sphingosine) of March_11_9_pos... Max. 3.e8 cps. 3.e8 2.5e Sphingosine 2 2.e8 +EMS: to min from Sample 1 (sphingosine) of March_11_9_pos_EMS_... 3.e Max. 3.1e6 cps. 1.5e8 2.5e6 2.e6 1.e8 1.5e6 1.e e e m/z, amu Time, min Cadenza CW-C18, 15 x 2 mm A: [water / C (1:.2 v/v)] containing 7mM ammonium formate B: [methanol / C (1:.2 v/v)] containing 5mM ammonium formate 7-9B (-1min), 9B (1-1.5min), 9-1B ( min), 1B (2.5-6min), 1-4B (6-6.5min), 4B (6.5-1min).2mL/min -.28mL/min (-6min),.2mL/min (6-1min) 48deg.C, 5uL (5ug / ml) Applied Biosystems QTrap, ESI Positive [M-2+] Courtesy of Jennifer Ro and Joe Williams Department of Evolution, Ecology, and rganismal Biology, hio State University - USA TI489E-IC2

30 o.ti49e LC-MS/MS Application for Sphingolipids (Sphingosine-1-P4) XIC of -EMS: to 38.9 amu from Sample 1 (S1P) of March_13_9_neg_EMS_... Max. 1.5e6 cps. 1.5e6 1.4e6 1.2e6 1.e6 8.e5 6.e5 4.e EMS: to min from Sample 1 (S1P) of March_13_9_neg_EMS_S1P.wiff e4 2.4e4 2.2e4 2.e4 1.8e4 1.6e4 1.4e4 1.2e4 1.e Max. 2.7e4 cps m/z, amu P + 3 Sphingosine-1-Phosphate 2.e Time, min Cadenza CW-C18, 15 x 2 mm A: [water / C (1:.2 v/v)] containing 7mM ammonium formate B: [methanol / C (1:.2 v/v)] containing 5mM ammonium formate 7-9B (-1min), 9B (1-1.5min), 9-1B ( min), 1B (2.5-6min), 1-4B (6-6.5min), 4B (6.5-1min).2mL/min -.28mL/min (-6min),.2mL/min (6-1min) 48deg.C, 5uL (3ug / ml) Applied Biosystems QTrap, ESI egative Courtesy of Jennifer Ro and Joe Williams Department of Evolution, Ecology, and rganismal Biology, hio State University - USA TI49E-IC2

31 o.ti491e LC-MS/MS Application for Sphingolipids (Sphinganine-1-P4) XIC of -EMS: to amu from Sample 2 (dhs1p) of March_13_9_neg_EMS... Max. 4.6e6 cps. 4.5e6 4.e6 3.5e6 3.e6 2.5e6 2.e6 1.5e6 1.e6 5.e EMS: to 4.86 min from Sample 2 (dhs1p) of March_13_9_neg_EMS_S1P m/z, amu Time, min 1.4e5 1.2e5 1.e5 8.e4 6.e4 4.e4 2.e P + 3 Sphinganine-1-Phosphate (Dihydrosphingosine-1-phosphate) Cadenza CW-C18, 15 x 2 mm A: [water / C (1:.2 v/v)] containing 7mM ammonium formate B: [methanol / C (1:.2 v/v)] containing 5mM ammonium formate 7-9B (-1min), 9B (1-1.5min), 9-1B ( min), 1B (2.5-6min), 1-4B (6-6.5min), 4B (6.5-1min).2mL/min -.28mL/min (-6min),.2mL/min (6-1min) 48deg.C, 5uL (5ug / ml) Applied Biosystems QTrap, ESI egative Courtesy of Jennifer Ro and Joe Williams Department of Evolution, Ecology, and rganismal Biology, hio State University - USA TI491E-IC2

32 o.ti493e LC-Q-TF-MS application for high-throughput non-targeted metabolomics with a hybrid DS column Column switching PLC waste PLC 1 2 QTF-MS take 1 µl urine centrifuge with.1 µm membrane filter umber of urine samples analyzed: up to 1, ea proteins depleted here! Time igh-throughput LC-MS analytical platform (~ 1 samples/day) Cadenza S-C18, 1 x 2 mm A:.1 formic acid in 5 acetonitrile B:.1 formic acid in 95 acetonitrile 1B (-3min), 1-9B (3-12min), 9-1B (12-14min).32mL/min, 4deg.C, 5-1uL Q-TF-MS, Switching time: 3min Ref) KISToday, Vol.2, o.1, 15-17, January 29 Courtesy of Man-o Choi, Ph.D. Life Sciences Division, Korea Institute of Science and Technology TI493E-IC21

33 o.ti515e LC-MS/MS application for saccharides Intensity, cps Fructose API5 (TAKARA BI IC. ) Glucose 1 pg/ul Time, min Area, counts Area, counts 3.4e6 3.e6 2.e6 1.e6. 4.8e6 4.e6 3.e6 2.e6 1.e6 y = 6.77e+3 x e+3 (r =.9998) ("1 / x" weighting) C2 D-glucose Concentration, pg/ul y = 9.68e+3 x (r =.999) ("1 / x" weighting) Concentration, pg/ul Unison UK-Amino, 15 x 2 mm acetonitrile / water = 9 / 1,.4 ml/min, 1 ul API5: ESI, MRM egative, Q1/Q3=179. / 89. 2C C 2 D-fructose Courtesy of TAKARA BI IC., Japan TI515E-ID28

34 o.ti516e LC-MS/MS application for saccharides Intensity, cps 4.6e4 4.e4 3.e4 2.e4 1.e4 fructose API5 (TAKARA BI IC. ) sucrose 1 pg/ul Area, counts. 3.2e6 3.e6 2.8e6 2.6e6 2.4e6 2.2e6 2.e6 1.8e6 1.6e6 1.4e6 1.2e6 1.e6 8.e5 6.e5 4.e5 2.e Time, min y = 6.42e+4 x ( r =.9995) ("1 / x" wei ghti ng) 2C C 2 D-fructose Concent r ation, ng/ml Area, counts 3.e6 2.8e6 2.6e6 2.4e6 2.2e6 2.e6 1.8e6 1.6e6 1.4e6 1.2e6 1.e6 8.e5 6.e5 4.e5 2.e5. y = 2.97e+4 x (r =.9984) ("1 / x" wei ghti ng) C2 C2 C 2 sucrose Concent r ation, ng/ml Unison UK-Amino, 15 x 2 mm acetonitrile / water = 83 / 17,.4 ml/min, 1 ul API5: ESI, MRM egative Q1/Q3: fructose 179. / 89., sucrose / 89. Courtesy of TAKARA BI IC., Japan TI516E-IE7

35 o.ti6e Analysis of Water-soluble Vitamins using LCMS (+):265.1 (+):17.2 (+):124.2(4.) (+):123.2(2.) (+):22.2(2.5) (+):678.6(8.) (+):377.2(4.) (+):245.1(2.5) (-):44.1(18.) C 3C 2 C2 C2 C C C2 2 + C3 S C2C2 thiamine pyridoxine nicotinic acid nicotinamide C2 3C C 3C 2CC2 3C 3C 3C C 2CC2C2 2CC2 C C Co+ C2C2 C3 C3 C2C2 C pantothenic acid C2C2C2 C3 C3 C3 C2C2C2 - C2C2CC2 C P 3C C2 C3 C3 cyanocobalamin min Shimadzu LCMS-22 (ESI, positive/negative, SIM) Scherzo SM-C18, 15 x 2 mm A: 5 mmol/l ammonium formate +.1 formic acid - water B: acetonitrile - 55 B (-1 min), B(1.1-2 min) 1, pg/ul, 1uL 7 C2C C 3C 3C 8 S 2 C2 9 CC2 C riboflavin biotin C C C C2C2 C folic acid Imtakt USA (info@imtaktusa.com) orth America Imtakt Corp./JAPA (info@imtakt.com) ther Countries TI6E-JF8

36 o.ti68e LC-MS(MRM) Application For Monosaccharides Intensity, cps 4.2e4 4.e4 3.8e4 3.6e4 3.4e4 3.2e4 3.e4 2.8e4 2.6e4 2.4e4 2.2e4 2.e4 1.8e4 1.6e4 1.4e4 1.2e4 1.e a b c d e f g (a) (b) (c) (d) (e) (f) (g) (h) (i) Max. 2.6e4 cps. Fucose Xylose -acetylgalactosamine -acetylglucosamine Mannose Galactose Glucose -acetylneuraminic acid -glycolylneuraminic acid Time, min h i Unison UK-Amino, 25 x 2 mm A: [.1 w/w ammonium acetate in water] B: [1 acetonitrile] B ( min), 89 B ( min), 89-3 B (17-26 min), 3 B ( 26-3 min), 91 B (3-4 min) 178 µl/min 55 deg.c, 1 µl (1 pg / µl in 91 B) Applied Biosystems QTrap 4; ESI egative; [M+C3C2] - precursor to [M-] - product for (a-g); and [M-] - parent to m/z 87 and 116 for h and i, respectively Ref) Rapid Communications in Mass Spectrometry Vol 24 Issue 11 (June 21) Courtesy of Loubna ammad, Dakota Derryberry, Yazen Jmeian, and Yehia Mechref METACyt Biochemical Analysis Center, Department of Chemistry, Indiana University 8 E. Kirkwood Ave, Bloomington, Indiana 4745 USA Imtakt USA (info@imtaktusa.com) orth America Imtakt Corp./JAPA (info@imtakt.com) ther Countries TI68E-JE18

37 o.ti645e LC-MS/MS Application: Simultaneous determination of phytotoxins in urine α-amanitin Phalloidin Colchicine Scopolamine Mesaconitine α-solanine Aconitine Jesaconitine α-chaconine ypaconitine Atropine Scherzo SM-C18, 15 x 2 mm 1mM ammonium formate / methanol = 1 / 9.2 ml/min, 4 deg.c, 1uL ESI-MS (MRM, positive) Ref.) Simultaneous Determination of phytotoxins in urine with LC/MS/MS Kouji Tachino, Michiko Fujiwara, Izumi Miura Courtesy of TACI Kouji, Yamaguchi Prefectual Institute of Public ealth and Environment, JAPA Imtakt USA (info@imtaktusa.com) orth America Imtakt Corp./JAPA (info@imtakt.com) ther Countries TI645E-KB22

38 o.ti652e Exploring different C18 stationary phases for metabolite profiling Best retention of polar compounds XBridge C18 (15 x 1. mm) 1. alanine 2.,-dimethylglycine 3. serine 4. fumaric acid 5. succinic acid 6. cysteine 7. oxaloacetic acid 8. Gly-Gly 9. malic acid 1. alpha-ketoglutarate 11. citric acid methylhippuric acid 13. gamma-d-glutamylglycine 14. salbutomal 15. AMP 16. ribo-avin 17. Phe-Gly-Phe-Gly 18. s-(p-nitrobenzyl)glutathione 19. s-(p-azidophenacyl)glutathione 2. Taurocholic acid 21. -beta-d-glucuronosyl-naphthol AS-BI 22. Leucine enkephalin 23. Arg-Pro-Pro-Gly-Phe (Bradykinin fragment 1-5) 24. Reserpine 25. GSSG 26. Arg-Lys-Asp-Val-Tyr (Thymopoietin II fragment 32-36) 27. Coenzyme A 28. Acetyl CoA 29. Ala-Ser-Thr-Thr-Thr-Asn-Tyr-Thr (peptide T) 3. Arg-Pro-ProGly-Phe-Ser-Pro-Phe (bradykinin fragment 1-8) 31. Tyr-Tyr-Tyr-Tyr-Tyr-Tyr Imtakt USA (info@imtaktusa.com) orth America Scherzo SM-C18, 15 x 2 mm A:.1 formic acid in water B:.1 formic acid in acetonitrile The linear gradient elution used started at 1 A (time -5 min) and finished at 1 B (35-4 min). 25 ul/min, room temp. ESI-MS in positive and negative ionization mode Ref.) Expanding Coverage of the Metabolome for Global Metabolite Profiling scar Yanes, Ralf Tautenhahn, Gary J. Patti, and Gary Siuzdak Analytical Chemistry, 211, 83(6), pp Courtesy of scar Yanes, Ph.D. Metabolomics Platform of the Spanish Biomedical Research Centre in Diabetes and Associated Metabolic Disorders (CIBERDEM), University Rovira i Virgili, SPAI Imtakt Corp./JAPA (info@imtakt.com) ther Countries TI652E-KD4

39 o.ti654e LC-MS/MS analysis of perfluorinated organic compounds (PFCs) in seawater PEpA (2ppt) MPFxA PFxA (53ppt) L-PFBS (<1ppt) PFPeA (<1ppt) MPFBA PFBA (<5ppt) MPFA PFA (4ppt) MPFA (I.S.) MPFA PFA (12ppt) MPFxS L-PFxS (<1ppt) Standard MPFUdA PFDA PFUdA (<1ppt) MPFDA PFDA (<1ppt) MPFS L-PFS (2ppt) PFxDA PFTeDA PFTrDA MPFDoA PFDoA L-PFDS Unison UK-C18, 25 x 2 mm A: 1mM ammonium acetate, B: acetonitrile 2-95B (-2min), 95B (2-28min), 2B (28-38min).2 ml/min (14.2MPa), 4 C 1uL ( <1-53ppt in seawater, 1ppb in surrogate or standard solution) ESI-MS/MS, negative Imtakt USA (info@imtaktusa.com) orth America Courtesy of YAGI Masahiro, Kobe Institute of ealth, JAPA Imtakt Corp./JAPA (info@imtakt.com) TI654E-KE2 ther Countries

40 o.ti655e LC-MS/MS analysis of nitrofurans metabolites (AZ, AMZ, AD) AMZ d > AZ d > > > 13.8 AMZ > AZ > > > AD 13C3 AD > > AMZ 3-amino-5-morpholinomethyl- 2-oxazolidinone > AZ 3-amino-2-oxazolidinone Cadenza CD-C18, 15 x 2 mm A:.1 Ac B: acetonitrile 2-8B (-15min), Runtime 19min.2 ml/min, 4 C 5uL (AZ, AMZ, AD 1. ppb, IS 5.ppb) ESI, MRM, positive Courtesy of MAEDA aoyuki, ASSCIATI F MEAT SCIECE & TECLGY ISTITUTE, JAPA Imtakt USA (info@imtaktusa.com) orth America 2 AD 1-aminohydantoin Imtakt Corp./JAPA (info@imtakt.com) ther Countries TI655E-KD14

41 o.ti656e LC-MS/MS analysis of clenbuterol (CBR) CBR d9 CBR 1 CBR 2 CBR 3 Cadenza CD-C18, 15 x 2 mm A:.1 formic acid in water B:.1 formic acid in acetonitrile 5-95B (-7min), 95B (7-1min) Runtime 15min.2 ml/min, 4 C 5uL (CBR.5ppb, IS.5ppb) ESI, MRM, positive 2 Cl Cl clenbuterol (CBR) C 3 C 3 C 3 Courtesy of MAEDA aoyuki, ASSCIATI F MEAT SCIECE & TECLGY ISTITUTE, JAPA Imtakt USA (info@imtaktusa.com) orth America Imtakt Corp./JAPA (info@imtakt.com) ther Countries TI656E-KD14

42 o.ti657e LC-MS/MS analysis of [leuco]malachite green (MG, LMG) MGd5 1: MRM of 4 Channels ES > e : MRM of 4 Channels ES > e MG 2: MRM of 4 Channels ES+ 337 > e4 1: MRM of 4 Channels ES > e : MRM of 4 Channels ES > e Time (min) malachite green (MG) Cadenza CD-C18 15 x 2 mm A: 1mM C4 / AC = 9 /1 B: acetonitrile -1B (-2min) 1B (2-3min) Runtime 4min.2 ml/min, 4 C 5uL (MG, LMG 1ppb, IS 25ppb) ESI, MRM, positive LMGd6 2: MRM of 4 Channels ES+ 337 > e LMG 2: MRM of 4 Channels ES > e : MRM of 4 Channels ES > e Time (min) leucomalachite green (LMG) Courtesy of MAEDA aoyuki, ASSCIATI F MEAT SCIECE & TECLGY ISTITUTE, JAPA Imtakt USA (info@imtaktusa.com) orth America Imtakt Corp./JAPA (info@imtakt.com) ther Countries TI657E-KD15

43 o.ti658e LC-MS/MS analysis of chloramphenicol (CP) RT: CP 1ppb CP L: 1.62E3 Base Peak m/z= F: - c ESI SRM ms [ , ] MS CP_11 1 Relative Abundance CPd5 L: 7.54E2 Base Peak m/z= F: - c ESI SRM ms [ , ] MS CP_11 L: 1.98E3 Base Peak m/z= F: - c ESI SRM ms [ , ] MS CP_ L: 2.22E3 Base Peak m/z= F: - c ESI SRM ms [ , ] MS CP_ Time (min) Cadenza CD-C18, 15 x 2 mm 1mM C4 / AC = 7 / 3 Runtime 15min.2 ml/min, 4 C 5uL (CP 1.ppb, IS 5.ppb) ESI, SRM, negative + - Cl Cl chloramphenicol (CP) Courtesy of MAEDA aoyuki, ASSCIATI F MEAT SCIECE & TECLGY ISTITUTE, JAPA Imtakt USA (info@imtaktusa.com) orth America Imtakt Corp./JAPA (info@imtakt.com) ther Countries TI658E-KD15

44 o.ti664e LC-MS analysis of phosphorus-containing amino acid type herbicides and their metabolites 6 glyphosate (m/z: ) 15 AMPA (metabolite) (m/z: 11.7) Intensity 4 2 P Intensity 1 5 P min min 4 glufosinate (m/z: ) 8 MPPA (metabolite) (m/z: ) Intensity 3 2 P 3C 2 Intensity 6 4 P 3C min min Intensity bialaphos (m/z: ) P 3C min 2 C3 C3 Scherzo SM-C18, 3 x 3 mm A:.1 C B: acetonitrile 5-3B ( - 3 min).5 ml/min 1uL (1ug/mL) ESI (Jet Stream), Q-Tof, SIM, negative mode Imtakt USA (info@imtaktusa.com) orth America Imtakt Corp./JAPA (info@imtakt.com) ther Countries TI664E-KE2

45 o.ti665e LC-ICP-MS analysis of Cr(III) and Cr(VI) Cr 6+ Cr 3+ Intensity (-) µg L -1 1 µg L -1 1 µg L -1 1 µg L µg L µg L Retention time (min) Scherzo SM-C18, 5 x 3 mm A: phosphoric acid / a (25mM, p 7) B: 1mM phosphoric acid B (-1min), -1B (1-4min), 1B (4-7min) 1. ml/min 15uL [ICP-MS] ELA DRC-e (PerkinElmer) m/z 52 Imtakt USA (info@imtaktusa.com) orth America Imtakt Corp./JAPA (info@imtakt.com) ther Countries TI665E-KE21

46 o.ti676e T Ultra-high throughput LC-MS/MS analysis of drugs in blood plasma T Area, counts T Cadenza CD-C18 T, 5 x 2 mm A:.1 C in water, B:.1 C in acetonitrile 2µL (2µL plasma + 5µL acetonitrile) Shimadzu UFLC - AB Sciex API5 (ESI positive, MRM) FD(nifedipine) 347 > 254, WAR(warfarin) 39 >163, DCL(diclofenac) 296 > 214, CBP(2-amino-5-chlorobenzophenone) 232 > 126 Imtakt USA (info@imtaktusa.com) orth America Courtesy of Dr. IKEARA Tatsuya, Shionogi Co., Ltd., JAPA Imtakt Corp./JAPA (info@imtakt.com) ther Countries TI676E-KG16

47 o.ti685e Peptide mapping using LC-FTMS RT: Relative Abundance B (-6 min).2 ml/min B (-6 min).7 ml/min Relative Abundance Time (min) Presto FF-C18, Cadenza CD-C18, 15 x 3 mm A: water / formic acid / TFA = 1 /.5 /.2 B: acetonitrile / formic acid /TFA = 1 /.5 /.2 5 deg.c, 2uL (tryptic digest of alpha-casein) FTMS (Thermo Scientific Exactive), ESI-positive (Full ms) Courtesy of Dr. KUWATA Keiko, Kyoto University, JAPA Imtakt USA (info@imtaktusa.com) orth America Imtakt Corp./JAPA (info@imtakt.com) ther Countries TI685E-K12