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1 7 7 Vol. 7 No Journal of Food Safety and Quality Jul., * 于泓, 胡青, 张甦, 孙健, 冯睿, 毛秀红, 季申 (, ) 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 ~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 , 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 : (14DZ ) Fund: Supported by Science and Technology Commission of Shanghai Municipality Project (14DZ ) * 通讯作者 :,,, [email protected] *Corresponding author: JI Shen, Chief Pharmacist, Ph.D Supervisor, Shanghai Institute for Food and Drug Control, No Zhangheng Road, Pudong New District, Shanghai , China. [email protected]

2 7, : 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], [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,, : , : 95.2%) (acipimox,, : ) (benfluorex, Sigma, : 087F0342) (benzafibrate,, : ) (pantethine,, : , : 79.3%) (fenofibrate,, : ) (furazabol,, : ) (fluvastatin,, : , : 97.0%) (ciprofibrate, Toronto Research Chemicals TRC, : 4-BLL-169-1, : 98.0%) (gemfibrozil,, : , : 99.5%) (lovastatin,, : , : 99.4%) (clofibrate Dr. Ehrenstorfer GmbH, : C , : 99.8%) (mevastatin, TRC, : 25-SSR-104-1, : 98.0%) (rravastatin,, : , : 96.7%) (probucol,, : ) (rosuvastatin, TRC, : 11-YM-77-1) (simvastatin,, : ) (nicotinic acid,, : , : 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, 实验方法 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): ev 21 (extracted ion chromatogram, EIC), 1, 1

3 Fig ( : 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 [M+H] , , , , (acipimox) C 6 H 6 O 3 N [M+H] , (benfluorex ) C 19 H 20 F 3 NO [M+H] , , , , (benzafibrate) C 19 H 20 ClNO [M+H] , , , , (pantethine) C 22 H 42 N 4 O 8 S [M+H] , , , , (fenofibrate) C 20 H 21 ClO [M+H] , , (furazabol) C 20 H 30 N 2 O [M+H] (fluvastatin) C 24 H 25 FNO [M+H] , , (ciprofibrate) C 13 H 14 Cl 2 O [M-H] , (gemfibrozil) C 15 H 22 O [M-H] (lovastatin) C 24 H 36 O [M+H] , , , , , , , (clofibrate) C 12 H 15 ClO [M+H] , , , , (mevastatin) C 23 H 34 O [M+H] , , , (rravastatin ) C 23 H 36 O [M+Na] , , (probucol) C 31 H 48 O 2 S [M-H] , , (rosuvastatin ) C 22 H 27 FN 3 O 6 S [M+H] , , (simvastatin) C 25 H 38 O [M+H] , , , , , , (nicotinic acid) C 6 H 5 NO [M+H] , , (ezetimibe) C 24 H 21 F 2 NO [M+H] , , , (dehydro lovastatin) (lovastatin hydroxy acid) C 24 H 34 O [M+H] , , , , C 24 H 38 O [M+Na] , , ,

4 7, : 样品前处理 (1) / /,,,,, 50 ml,, ( 500 W 50 khz)30 min,,, r/min 10 min, (2), 50 ml, 30 ml( : 50% ), ( 500 W 50kHz)30 min,,, r/min 10 min, 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) 标准谱库的建立 Mass Hunter PCDL Manager( Agilent ), (collision-induced dissociation, CID),, 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, , 2.2.3,, 检出限,, ,, 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) 重复性试验 2.2.2,, 21,, 6, 21 (n=6), 3.4 稳定性试验, h, 21

5 RSD 0.54%~5.44% h 3.5 实际样品测定, 96 1 ( ), 4 讨论 (S/N=3),,,,,, ( 18),,,,,,,, 10 [3],,, 21,, [15] 参考文献 [1],. 185 [J]., 2016, 27(2): Zhao XL, Wang M. Analysis of 185 cases of statin-induced ADR in lipid-lowering therapy in our hospital [J]. J China Pharm, 2016, 27(2): [2] Gaist D, Rodríguez LAG, Huerta C, et al. Lipid-lowering drugs and risk of myopathy: a population-based follow-up study [J]. Epidemiology, 2001, 12(5): [3] 喆,,,. [J]., 2008, 28(12): Zhang Z, GaoQ, Che BQ, et al. HPLC-MS/MS determination of ten chemicales in traditional Chinese medicine and health food [J]. Chin J Pharm Anal, 2008, 28(12): [4],,,. Q-Orbitrap [J]., 2015, 35(8): Jiang SY, Guo CC, Shi F, et al. Application of Q-Orbitrap high resolution mass spectrometry to the detection of illegal adulterated antihyperlipidemics in herbal medicines and dietary supplements [J]. Chin J Pharm Anal, 2015, 35(8): [5],,,. UPLC-MS/MS 37 [J]., 2016, 36(5): Gong X, Lu L, Feng YL, et al. Determination of 37 drugs illegally added in weight-losing, lipid-lowering and laxative health foods by UPLC-MS/MS [J]. Chin J Pharm Anal, 2016, 36(5): [6],, 甦, [J]., 2009, 40(2): Hu Q, Cui YL, Zhang S, et al. Determination of acetic acid in compound amino acids injection by ion chromatography [J]. Chin J Pharm, 2009, 40(2): [7] 甦,,,. 48 [J]., 2015, 37(3): Zhang S, Hu Q, Sun J, et al. Simultaneous determination of 48 antibiotics in Chinese patent medicines and health foods by LC-ITMS [J]. Chin Tradit Pat Med, 2015, 37(3): [8], 甦,,. LC MS [J]., 2011, 21(8): Hu Q, Zhang S, Jian LH, et al. Identification of sibutramine and its derivatives added illegally into functional food by LC-MS [J]. Chin J Heal Lab Tech, 2011, 21(8): [9],, 甦,. [J]., 2009, 31(10): Hu Q, Cui YL, Zhang S, et al. Detection of chemical drugs added illegally into analgesic-antipyretic traditional Chinese medicines and health food by LC/MS [J]. Chin Tradit Pat Med, 2009, 31(10): [10],, 甦, [J]., 2008, 30(7): Hu Q, Cui YL, Zhang S, et al. 15 adulterated chemicals in antihypertension traditional Chinese patent medicine and health food by LC-ITMS [J]. Chin Tradit Pat Med, 2008, 30(7): [11],,, [J]., 2008, 28(8): Wang K, Hu Q, Cui YL, et al. Detection of 28 chemical drugs added illegally into slimming Traditonal Chinese Medicines and health food by liquid chromatography-ion trap mass spectrometry method [J]. Chin J Pharm Anal, 2008, 30(7): [12],,, [J]., 2008, 28(12): Hu Q, Cui YL, Wang K, et al. Detection of 16 anti-inflammatory and

6 7, : anti-histamine chemical drugs added illegally into traditional Chinese medicine and health food by liquid chromatography-ion trap mass spectrometry method [J]. Chin J Pharm Anal, 2008, 28(12): [13],,,. Q-TOF-MS [J]., 2014, (4): Sun J, Yu H, Hu Q, et al. Identification of a new sildenafil analogue based on Q-TOF-MS [J]. Acta Pharm Sin, 2014, (4): [14],, 甦, [J]., 2013, 48(15): Yu H, Hu Q, Zhang S, et al. Qualitative and quantitative analysis of 31 PDE-5 inhibitors in herbal products and dietary supplements by UPLC/Q-TOF-MS [J]. Chin Pharm J, 2013, 48(15): [15] Wang Z, Wu C, Wang G, et al. Novel strategy for the determination of illegal adulterants in health foods and herbal medicines using high-performance liquid chromatography with high -resolution mass spectrometry [J]. J Sep Sci, 2015, 38(6): 作者简介 ( 责任编辑 : 姚菲 ) 于泓, 药师, 主要研究方向为中药和保健食品质量和安全性检测 [email protected] 季申, 博士, 主任药师, 主要研究方向为中药质量标准及中药安全性研究 [email protected]