7 7 Vol. 7 No. 7 2016 7 Journal of Food Safety and Quality Jul., 2016 - - 21 * 于泓, 胡青, 张甦, 孙健, 冯睿, 毛秀红, 季申 (, 201203) 摘要 : 目的 - - (ultra performance liquid chromatography/ quadrupole-time-of-flight mass spectrometry, UPLC/Q-TOF-MS) 21 方法, Agilent Poroshell 120 SB-C 18, 0.1%, 0.4 ml/min; Q-TOF-MS, (electronic spray ion, ESI), 结果 21 0.05~25 ng 96, 1, 结论, 关键词 : - - ; ; ; ; Determination of 21 kinds of lipid-lowering drugs in herbal products and dietary supplements by ultra performance liquid chromatography/ quadrupole-time-of-flight mass spectrometry YU Hong, HU Qing, ZHANG Su, SUN Jian, FENG Rui, MAO Xiu-Hong, JI Shen * (Shanghai Institute for Food and Drug Control, Shanghai 201203, China) ABSTRACT: Objective To establish a method for determination of 21 kinds of lipid-lowering drugs in herbal products and dietary supplements by ultra performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS). Methods The samples were processed by ultrasonic extraction with methanol, and separated by Agilent Poroshell 120 SB-C 18 column with 0.1% formic acid and acetonitrile as mobile phase at the flow rate of 0.4 ml/min for gradient elution. Q-TOF-MS and electronic spray ion (ESI) both in positive and negative ionization mode were used in mass. Qualitative analysis was based on retention time, accurate mass, the elemental compositions and the product ions. Results The limits of detection (LOD) of 21 kinds of lipid-lowering drugs were 0.05~25 ng. Ninety-six batches of commercial herbal products and dietary supplements were detected, only 1 batch of tablet los simvastatin was detected, and the rest were not checked out. Conclusion The established method is sensitive, accurate and efficient, which is suitable for rapid determination of 21 kinds of lipid-lowering drugs in 基金项目 : (14DZ2294000) Fund: Supported by Science and Technology Commission of Shanghai Municipality Project (14DZ2294000) * 通讯作者 :,,, E-mail: jishen2013@163.com *Corresponding author: JI Shen, Chief Pharmacist, Ph.D Supervisor, Shanghai Institute for Food and Drug Control, No. 1500 Zhangheng Road, Pudong New District, Shanghai 201203, China. E-mail: jishen2013@163.com
7, : - - 21 2705 herbal products and dietary supplements. KEY WORDS: ultra performance liquid chromatography/quadrupole-time-of-flight mass spectrometry; lipid-lowering drugs; herbal products; dietary supplements; illegal additives 1 引言,,,,, [1,2] [2010]114 1 [2016]28 8, 6,, [3-6], - - 31 408 [6-12], - (ultra performance liquid chromatography/ quadrupole-time-of-flight mass spectrometry, UPLC/Q- TOF-MS) [13,14],, UPLC/Q-TOF-MS 21,, ( ) 2 材料与方法 2.1 仪器与材料 Agilent 1290 Infinity, Agilent 6530 Q-TOF ( Agilent ); Millipore Milli-Q Gradient A10 ( Millipore ); Sartorius CP225D ( Sartorius ); ( ) (LC-MS, Merck ), (HPLC, Merck ), (LC-MS, Fluka ); Agilent : (atorvastatin,, : 100590-200501, : 95.2%) (acipimox,, : 100750-200601) (benfluorex, Sigma, : 087F0342) (benzafibrate,, : 100732-200501) (pantethine,, : 100917-200701, : 79.3%) (fenofibrate,, : 100733-200401) (furazabol,, : 100387-200401) (fluvastatin,, : 100800-200701, : 97.0%) (ciprofibrate, Toronto Research Chemicals TRC, : 4-BLL-169-1, : 98.0%) (gemfibrozil,, : 100284-200602, : 99.5%) (lovastatin,, : 100600-200301, : 99.4%) (clofibrate Dr. Ehrenstorfer GmbH, : C11682000, : 99.8%) (mevastatin, TRC, : 25-SSR-104-1, : 98.0%) (rravastatin,, : 100689-200401, : 96.7%) (probucol,, : 100560-200301) (rosuvastatin, TRC, : 11-YM-77-1) (simvastatin,, : 100601-200502) (nicotinic acid,, : 100434-201112, : 99.9%) (ezetimibe, TRC, : 11-ANR-58-1, : 98.0%) (dehydro lovastatin, TRC, : 54-vkv-11-1, : 98.0%) (Lovastatin Hydroxy Acid, TRC : 18-MVI-65-2, : 96.0%) : ( ) ( ) ( ), 29, 10, 57 2.2 实验方法 2.2.1 仪器条件 (1) : Agilent Poroshell 120 SB-C 18 (3.0 mm 75 mm, 2.7 μm); : 30 C; : 1~5 μl; : A 0.1% -, B ; : 0~10 min: 5%~98%B; 10~13 min: 98%B; 13~14 min: 98%~5%B; 14~17 min: 5%B : 0.4 ml/min (2) : : ; m/z: 100~1200; : (N 2 ): 35 psi; (N 2 ): 12 L/min, 350 ; (N 2 ): 12 L/min, 450 ; 3.5 kv; 175 V; (collision-induced dissociation, CID): 10 20 40 ev 21 (extracted ion chromatogram, EIC), 1, 1
2706 7 Fig. 1 1 21 ( : 10 μg/ml) Extracted ion chromatogram of 21 kinds of compounds (mass window: 10 μg/ml) Table 1 表 1 21 种化合物的分子式 精确分子量 保留时间 碰撞电压和碎片离子 Molecular formula, precursor ions, retention time, CID pressures and fragment ions of 21 kinds of compounds (m/z) (min) CID (ev) 1 (atorvastatin) C 33 H 35 FN 2 O 5 559.2603[M+H]+ 7.10 10 466.2005,440.2216,292.1481,276.1171,250.1015 2 (acipimox) C 6 H 6 O 3 N 2 155.0451[M+H]+ 1.33 10 137.0349,109.0402 3 (benfluorex ) C 19 H 20 F 3 NO 2 352.1519[M+H]+ 5.87 20 230.1147,187.0726,159.0411,149.0593,105.0334 4 (benzafibrate) C 19 H 20 ClNO 4 362.1071[M+H]+ 6.33 10 316.1065,276.0744,161.0925,138.9911,121.0617 5 (pantethine) C 22 H 42 N 4 O 8 S 2 555.2517[M+H]+ 3.10 10 537.2407,425.1882,295.1250,147.0585,118.0319 6 (fenofibrate) C 20 H 21 ClO 4 361.1201[M+H]+ 9.40 10 233.0389,138.9970,121.0308 7 (furazabol) C 20 H 30 N 2 O 2 331.238[M+H]+ 8.43 40 119.0833 8 (fluvastatin) C 24 H 25 FNO 4 412.1919[M+H]+ 7.01 10 394.1774,266.1318,224.0866 9 (ciprofibrate) C 13 H 14 Cl 2 O 3 287.0247[M-H]- 7.10 10 200.9861,85.0296 10 (gemfibrozil) C 15 H 22 O 3 249.1496[M-H]- 8.21 10 121.0653 11 (lovastatin) C 24 H 36 O 5 405.2636[M+H]+ 8.41 5 303.1940, 285.1836, 267.1741, 243.1737, 225.1637, 199.1476, 173.1315, 143.0848 12 (clofibrate) C 12 H 15 ClO 3 243.0782[M+H]+ 8.21 10 197.0337, 169.0400, 141.0102, 115.0718, 59.0485 13 (mevastatin) C 23 H 34 O 5 391.2479[M+H]+ 8.03 10 229.1579, 211.1475, 185.1318, 159.1161 14 (rravastatin ) C 23 H 36 O 7 447.2353[M+Na]+ 4.95 20 429.2209, 327.1548, 143.0849 15 (probucol) C 31 H 48 O 2 S 2 515.3023[M-H]- 12.11 10 277.1624, 236.1239, 211.1004 16 (rosuvastatin ) C 22 H 27 FN 3 O 6 S 482.1756[M+H]+ 5.93 20 300.1496, 270.13959, 258.1390 17 (simvastatin) C 25 H 38 O 5 419.2792[M+H]+ 8.94 10 303.1953, 285.1854, 267.1732, 243.1735, 255.1633, 199.1478, 173.1322 18 (nicotinic acid) C 6 H 5 NO 2 124.0393[M+H]+ 1.20 20 80.0495, 78.0339, 53.0389 19 (ezetimibe) C 24 H 21 F 2 NO 3 410.156[M+H]+ 6.71 10 392.1449, 161.0594, 133.0648, 105.0700 20 21 (dehydro lovastatin) (lovastatin hydroxy acid) C 24 H 34 O 4 387.253[M+H]+ 9.59 10 285.1833, 239.1785, 199.1470, 173.1313, 143.0856 C 24 H 38 O 6 445.2561[M+Na]+ 7.54 20 319.1900, 101.0604, 85.0291, 59.0137
7, : - - 21 2707 2.2.2 样品前处理 (1) / /,,,,, 50 ml,, ( 500 W 50 khz)30 min,,, 14000 r/min 10 min, (2), 50 ml, 30 ml( : 50% ), ( 500 W 50kHz)30 min,,, 14000 r/min 10 min, 2.2.3 标准溶液的配制 20 mg, 20 ml,, ( 1 mg/ml) 1 ml, 100 ml,, ( 10 μg/ml), ( 1~1000 ng/ml) 2.2.4 标准谱库的建立 21 21 Mass Hunter PCDL Manager( Agilent ), (collision-induced dissociation, CID),, 1 2.2.5 检出结果判定,, 10 μg/ml, 0.5 min : (1) <5 μg/ml;(2) (Rt<0.2 min); (3), 3 结果与分析 3.1 专属性试验 21, : ( ) ( ), 2.2.2, 2.2.1, 2.2.4 21, 2.2.3,, 21 21 3.2 检出限,, 2.2.2 2.2.4,, 0.05~25 ng, 2, 1 Table 1 表 2 21 种化合物的检出限 (n=6) Limits of detection (LODs) of 21 kinds of compounds (n=6) NO. LOD(ng) LOD(ng) 1 0.25 0.25 2 2.5 2.5 3 0.05 0.05 4 0.25 0.25 5 0.05 0.05 6 0.05 0.05 7 2.5 2.5 8 0.5 0.5 9 0.5 0.5 10 2.5 2.5 11 0.05 0.05 12 25 25 13 0.05 0.05 14 0.5 0.5 15 2.5 2.5 16 0.05 0.05 17 0.05 0.05 18 0.25 2.5 19 0.25 0.25 20 0.25 0.25 21 0.5 0.5 3.3 重复性试验 2.2.2,, 21,, 6, 21 (n=6), 3.4 稳定性试验, 1 2 4 8 16 24 h, 21
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