Hydrocortisone Sodium Phosphate 3.5 1.0 0.5 1 μg/ml 3.0 2.5 25 ng/ml 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 min 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 min Fig.1 Chromatograms of Hydrocortisone Sodium Phosphate Instrument : Shimadzu NexeraXR+ LCMS2020 Column : Mastro TM C18(100 mml. 2.1 mmi.d., 3 µm) Typical C18(100 mml. 2.1 mmi.d., 3 µm) Flow Rate : 0.2 ml/min Detection : MS (SIM mode) Mobile Phase : A: 0.1% Formic Acid in Water B: 0.1% Formic Acid in MeCN 0.01 min ポンプ B.Conc 10% 5.00 min ポンプ B.Conc 80% 5.01 min ポンプ B.Conc 10% 6.50 min ポンプ B.Conc 10% Column Temp. : 40 Injection Vol. : 2 µl
Adenosine Triphosphate Adenosine Monophosphate 0.0 1.0 2.0 3.0 4.0 5.0 6.0 min 0.0 1.0 2.0 3.0 4.0 5.0 6.0 min Fig.2 Chromatograms of Nucleotides Peak Asymmetry Peak 1 Peak 2 1.39 1.14 5.10 1.49 Instrument Column Mobile Phase Flow Rate : Shimadzu Nexera : Mastro TM C18(100 mml. 2.1 mmi.d., 3 µm) Typical C18 (100 mml. 2.1 mmi.d., 3 µm) : 10 mm Ammonium Formate in Water : MeCN=99:1 : 0.2 ml/min Column Temp. : 40 Detection : SPD-M20A at (254 nm) Injection Vol. : 1 µl Fig.3
Fig.4 Chromatograms of Sugar Phospahates and Nucleotides Instruments : Shimadzu Nexera/LCMS-8050 Column : Mastro SP (2.1 mmi.d X 100 mml.,5 μm) Mobile phase A : 10 mm Ammonium acetate/mecn=90/10 B : 50 mm Ammonium acetate/mecn=80/20 Gradient B conc. : 0% B (0-4.0 min) 60% B (9.0 min) 100%B (11.0-20.0 min) 0%B (20.1-25.0 min) Oven Temp. : 40 Injection vol. : 3 µl Sample : 2.5 µm STD Mix T.Flow : 0.5 ml/min Nebulizing gas : 3.0 L/min Drying gas : 12 L/min DL Temp. : 250 BH Temp. : 400 Injection vol. : 3 µl セグメン ト#1 21,873 79,876 210,366 151,006 Methionine sulfone 2-Morpholinoethansulfonic acid Glucose 6-phosphate Ribose 5phosphate 180.10>79.15(-)@1 (1) 194.10>80.15(-)@2 (1) 259.00>97.10(-)@3 (1) 229.00>97.10(-)@4 (1) 164,146 289.00>97.10(-)@5 (1) Sedoheptulose 7-phosphate 170,327 40,854 97,411 69,439 Fructose 6-phosphate Glycerol 3-phosphate Glucose 1-phosphate Glyceroaldehyde 3-phosphate 259.00>97.10(-)@6 (1) 170.90>79.10(-)@7 (1) 258.90>79.10(-)@8 (1) 169.00>97.10(-)@9 (1) 26,920 198.90>97.10(-)@10 (1) Erythrose 4-phosphate 151,905 26,289 67,224 82,196 Ribulose 5-phosphate CMP Dihydroxyacetonephosphate UMP 229.00>97.10(-)@11 (1) 322.00>79.10(-)@12 (1) 169.00>97.10(-)@13 (1) 322.90>79.10(-)@14 (1) 57,987 361.90>79.10(-)@15 (1) GMP 129,243 63,354 193,379 373,078 23,121 24,843 58,584 86,086 47,954 61,741 63,139 173,838 18,372 11,988 28,845 24,690 51,203 31,425 TMP AMP cgmp camp CDP GDP UDP 3-Phosphoglycerate/2-Phosphoglycerate Fructose 1,6-bisphosphate ADP TDP Phosphoenolpyruvate CTP GTP UTP ATP TTP NAD 320.90>195.05(-)@16 (1) 346.00>79.10(-)@17 (1) 343.90>150.10(-)@18 (1) 327.90>134.20(-)@19 (1) 402.00>158.90(-)@20 (1) 441.90>159.05(-)@21 (1) 403.00>159.05(-)@22 (1) 185.10>97.10(-)@23 (1) 339.10>97.10(-)@24 (1) 425.90>159.00(-)@25 (1) 401.00>159.00(-)@26 (1) 167.10>79.10(-)@27 (1) 481.90>159.00(-)@28 (1) 521.90>159.00(-)@29 (1) 482.90>159.00(-)@30 (1) 505.90>159.05(-)@31 (1) 481.10>159.00(-)@32 (1) 662.00>539.95(-)@33 (1) 15,237 742.20>619.90(-)@34 (1) NADP 36,660 6,604 780 10,059 NADH NADPH Succinyl coenzyme A Acetyl coenzyme A 664.00>408.00(-)@35 (1) 744.20>408.15(-)@36 (1) 866.00>408.00(-)@37 (1) 808.00>408.00(-)@38 (1) 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 min Fig.5 Chromatograms of Sugar Phospahates and Nucleotides Instruments : Shimadzu Nexera/LCMS-8050 Column : Mastro SP (2.1 mmi.d.x 100 mml.,5 µm) Mobile phase A : 10 mm NH4HCO3/ MeCN=90/10 B : 20 mm NH4HCO3/ MeCN=70/30 Gradient B conc. : 0% (0-3 min) 100% (13.0-20.0 min) 0% (20.1-25.0 min) Oven Temp. : 40 Injection vol. : 2 µl Sample : 2.5 µm STD Mix Nebulizer Gas : 2.0 L/min Drying Gas : 10 L/min Heating Gas : 10 L/min Interface Temp. : 300 DL Temp. : 250 HB Temp. : 400 Dwell Time : 10 ms Pause Time : 1 ms Ionization mode : ESI/negative 注 ) アルカリ 条 件 下 ではカラム 寿 命 が 短 くなる 可 能 性 があります
3-ADON 15-ADON Fig.6 Chromatograms of 3-ADON/15-ADON,FB1 and FB2/FB3 using Mastro PFP,C18 and Typical PFP columns
NIV DON FUX NEO 15-ADON 3-ADON DAS HT-2 FB1 β-zal α-zal T-2 FB3 FB2 β-zel α-zel ZEN Fig.7 Chromatograms of 17 Mycotoxins Instruments Column : Nexera/LCMS-8060 : Mastro PFP (2.1 mmi.d.x 100 mml., 3 µm) Time program: B20%(0min)-40%(1-1.5min)-50%(1.51-5.5min) -65%(5.51min)-70%(9.5min)-95%(9.51-12min) Flow : 0.3 ml/min Mobile phase A: 10 mm Ammonium acetate in Water B: 2% Acetic acid in MeOH Sample : 50 ng/ml STD Mix