超高效液相色谱-串联质谱法同时测定保健食品中15种非法添加降糖化学药物
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摘要
目的建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)同时测定保健食品中15种非法添加降糖化学药物的方法。方法样品采用乙腈超声提取,采用UPLC-MS/MS进行检测,以Atlantis?dC15 (2.1 mm×150 mm, 3μm)为色谱柱, 0.1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱程序为0～3min,90%A;3～15min,90%～60%A;15～28min,60%～0%A;28～29min, 0%A; 29～30 min, 0～90%A; 30～31 min, 90%A);体积流量0.3 mL/min,柱温40℃,进样体积5μL。选择ESI离子源,多反应监测模式(multiple reaction monitoring, MRM),通过比较MRM通道中样品峰与对照品峰的分子离子峰,二级碎片离子峰等信息确定添加的化学药。结果 15种化学药物的线性范围宽,相关性好,r≥0.995,相对标准偏差(relative standard deviation, RSD)为1.58%～5.12%(n=6);加标浓度为20、100、200 ng/mL时,回收率为81.0%～120.1%,检出限为0.010～0.120μg/g,定量限在0.034～0.39μg/g。应用该方法对48批样品进行了检测,其中有2批次样品中检出盐酸苯乙双胍和格列本脲。结论本方法专属性强,灵敏度高,适合用于保健食品中非法添加15种降糖化学药物的快速检测。
Objective To establish a method for simultaneous determination of 15 kinds of illegally added hypoglycemic chemicals in health foods by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). Methods The sample was ultrasonically extracted with acetonitrile and detected by UPLC-MS/MS, using Atlantis?dC15(2.1 mm × 150 mm, 3 μm) as the column and 0.1% aqueous formic acid(A)-acetonitrile(B) as mobile phase. The gradient elution program was 0-3 min, 90%A; 3-15 min, 90%-60%A; 15-28 min, 60%-0%A; 28-29 min, 0%A; 29-30 min, 0～90%A; 30-31 min, 90%A. The volume flow rate was 0.3 mL/min,the column temperature was 40 ℃, and the injection volume was 5 μL. Results The linear range of 15 chemical drugs was wide and the correlation was good, r≥0.995, and the relative standard deviation(RSD) was 1.58%-5.12%(n=6). The recoveries at 3 spiked(20, 100, 200 ng/mL) were 81.0%-120.1%, the limits of detection were 0.010-0.120μg/g, and the limits of quantification were 0.034-0.39 μg/g. The 48 batches of samples were tested by this method,and phenformin hydrochloride and glibenclamide were detected in 2 batches. Conclusion This method has strong specificity and high sensitivity, which is suitable for rapid detection of illegally adding 15 kinds of hypoglycemic chemicals in health foods.
Objective To establish a method for simultaneous determination of 15 kinds of illegally added hypoglycemic chemicals in health foods by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). Methods The sample was ultrasonically extracted with acetonitrile and detected by UPLC-MS/MS, using Atlantis?dC15(2.1 mm × 150 mm, 3 μm) as the column and 0.1% aqueous formic acid(A)-acetonitrile(B) as mobile phase. The gradient elution program was 0-3 min, 90%A; 3-15 min, 90%-60%A; 15-28 min, 60%-0%A; 28-29 min, 0%A; 29-30 min, 0～90%A; 30-31 min, 90%A. The volume flow rate was 0.3 mL/min,the column temperature was 40 ℃, and the injection volume was 5 μL. Results The linear range of 15 chemical drugs was wide and the correlation was good, r≥0.995, and the relative standard deviation(RSD) was 1.58%-5.12%(n=6). The recoveries at 3 spiked(20, 100, 200 ng/mL) were 81.0%-120.1%, the limits of detection were 0.010-0.120μg/g, and the limits of quantification were 0.034-0.39 μg/g. The 48 batches of samples were tested by this method,and phenformin hydrochloride and glibenclamide were detected in 2 batches. Conclusion This method has strong specificity and high sensitivity, which is suitable for rapid detection of illegally adding 15 kinds of hypoglycemic chemicals in health foods.
引文
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[8]殷帅,吴公平,雷玉萍.液质联用法检测中成药和保健食品中添加辅助降血糖类药品的研究[J].药物分析杂志,2010,30(12):2400-2403.Yin S,Wu GP,Lei YP.HPLC-MS/MS determination of auxiliary hypoglycemic agents in traditional Chinese medicine and healthy foods[J].Chin J Pharm Anal,2010,30(12):2400-2403.
[9]吴杨,张圆,闵春艳,等.辅助降血糖类保健食品非法添加化学物质检测方法研究[J].中国执业药师,2014,11(10):11-14.Wu Y,Zhang Y,Min CY,et al.Study on the detection method for illegally added chemicals in auxiliary hypoglycemic health food[J].China Licens Pharm,2014,11(10):11-14.
[10]张喆,高青,余倩,等.中药制剂及保健品中违禁添加9种化学降糖药的HPLC-MS/MS定性检测[J].中国医药工业杂志,2007,38(1):39-43.Zhang Z,Gao Q,Yu Q,et al.Detection of 9 anti-diabetic agens in traditional chinese medicines and health care products by HPLC-MS/MS[J].Chin J Pharm,2007,38(1):39-43.
[11]隋海山,王立娟,戚威.HPLC-MS/MS法检测中成药和食品中违法添加的10种降糖类药物[J].中国医药指南,2016,22(1):36-37.Sui HS,Wang LJ,Qi W.Detection of anti-diabetic agents in chinese patent medicines and food by HPLC-MS/MS[J].Guid China Med,2016,22(1):36-37.
[12]罗疆南,曹玲,谭力,等.HPLC-MS/MS检测中药及保健品中添加的11种降糖类化学药物[J].中国药学杂志,2010,45(11):68-70.Lu JN,Cao L,Tan L,et al.Determination of 11 anti-diabetics mixed illegally into traditional Chinese medicines and health care products by HPLC-MS/MS[J].Chin Pharm J,2010,45(11):68-70.
[13]黄湘鹭,王静文,曹进,等.降血糖类保健食品中非法添加的8种药物检测及实例分析[J].食品科学,2014,35(10):149-152.Huang XL,Wang JW,Cao J,et al.Determination of 8 drugs in antidiabetic supplements:A case study[J].Food Sci,2014,35(10):149-152.
[14]魏清芳,王嘉林.液相色谱-串联四极杆质谱测定中药及保健品中非法添加8种化学降糖药[J].中国药事,2011,25(1):70-72.Wei QF,Wang JL.Detection of 8 kinds of antidiabetics illegally added in traditional chinese medicinal preparations and health foods by the liquid chromatography tandem with quadrupole mass spectrometry method[J].Chin Pharm Aff,2011,25(1):70-72.
[15]朱峰,阮丽萍,马永建,等.超高效液相色谱-串联质谱联用法同时检测降糖类和减肥类保健品中20种非法添加的化学降糖药物[J].色谱,2014,32(1):13-20.Zhu F,Ruan LP,Ma YJ,et al.Simultaneous detecmination of 20 illeyally added anti-diabetic chemical components in hypoglycemic and weight-reducing health foods by ultra-high performance liquid chromatography tandem mass spectrometry[J].Chin J Chromatogr,2014,32(1):13-20.
[2]李荣,卫星华,董靖.HPLC-DAD法测定中成药及保健食品中非法添加16种降糖类化学药物的研究[J].辽宁中医杂志,2018,45(5):1022-1025.Li R,Wei XH,Dong J.Determination of 16 antidiabetic agents illegally added in traditional chinese medicines and health food by HPLC-DAD[J].Liaoning J TCM,2018,45(5):1022-1025.
[3]励炯,曹青文,王姣斐,等.基于实心核颗粒色谱技术结合HPLC法快速测定保健食品中非法添加13种降糖化学成分[J].中草药,2017,48(13):2666-2669.Li J,Cao QW,Wang JF,et al.Detection of 13 anti-diabetic drugs illegally added in health food by HPLC with solid nuclear particle chromatographic column[J].Trad Herb Drug,2017,48(13):2666-2669.
[4]尚士博,安康,何漾,等.高效液相色谱法同时测定泡茶中的4种非法添加化学药物[J].食品安全质量检测学报,2018,9(16):4296-4300.Shang SB,An K,He Y,et al.Simultaneous determination of 4 kinds of illegal added of chemical drugs in teabags by high performance liquid chromatography[J].J Food Saf Qual,2018,9(16):4296-4300.
[5]朱俐,冯雪,尹利辉.离子迁移谱法快速筛查保健食品中非法添加降糖类药品[J].分析科学学报,2018,34(2):165-169.Zhu L,Feng X,Yin LH.Rapid screening of illegally added hypoglycemic drugs in health foods by ion migration spectrometry[J].J Anal Sci,2018,34(2):165-169.
[6]Fernandes RS,Costa FSL,Valderrama P,et al.Non-destructive detection of adulterated tablets of glibenclamide using NIR and solid-phase fluorescence spectroscopy and chemometric methods[J].J Pharm Biomed Anal,2012,(66):85-90.
[7]马双成,魏锋.保健食品安全性检测[M].北京:人民卫生出版社,2011.Ma SC,Wei F.Safety inspection of health food[M].Beijing:People's Medical Publishing House,2011.
[8]殷帅,吴公平,雷玉萍.液质联用法检测中成药和保健食品中添加辅助降血糖类药品的研究[J].药物分析杂志,2010,30(12):2400-2403.Yin S,Wu GP,Lei YP.HPLC-MS/MS determination of auxiliary hypoglycemic agents in traditional Chinese medicine and healthy foods[J].Chin J Pharm Anal,2010,30(12):2400-2403.
[9]吴杨,张圆,闵春艳,等.辅助降血糖类保健食品非法添加化学物质检测方法研究[J].中国执业药师,2014,11(10):11-14.Wu Y,Zhang Y,Min CY,et al.Study on the detection method for illegally added chemicals in auxiliary hypoglycemic health food[J].China Licens Pharm,2014,11(10):11-14.
[10]张喆,高青,余倩,等.中药制剂及保健品中违禁添加9种化学降糖药的HPLC-MS/MS定性检测[J].中国医药工业杂志,2007,38(1):39-43.Zhang Z,Gao Q,Yu Q,et al.Detection of 9 anti-diabetic agens in traditional chinese medicines and health care products by HPLC-MS/MS[J].Chin J Pharm,2007,38(1):39-43.
[11]隋海山,王立娟,戚威.HPLC-MS/MS法检测中成药和食品中违法添加的10种降糖类药物[J].中国医药指南,2016,22(1):36-37.Sui HS,Wang LJ,Qi W.Detection of anti-diabetic agents in chinese patent medicines and food by HPLC-MS/MS[J].Guid China Med,2016,22(1):36-37.
[12]罗疆南,曹玲,谭力,等.HPLC-MS/MS检测中药及保健品中添加的11种降糖类化学药物[J].中国药学杂志,2010,45(11):68-70.Lu JN,Cao L,Tan L,et al.Determination of 11 anti-diabetics mixed illegally into traditional Chinese medicines and health care products by HPLC-MS/MS[J].Chin Pharm J,2010,45(11):68-70.
[13]黄湘鹭,王静文,曹进,等.降血糖类保健食品中非法添加的8种药物检测及实例分析[J].食品科学,2014,35(10):149-152.Huang XL,Wang JW,Cao J,et al.Determination of 8 drugs in antidiabetic supplements:A case study[J].Food Sci,2014,35(10):149-152.
[14]魏清芳,王嘉林.液相色谱-串联四极杆质谱测定中药及保健品中非法添加8种化学降糖药[J].中国药事,2011,25(1):70-72.Wei QF,Wang JL.Detection of 8 kinds of antidiabetics illegally added in traditional chinese medicinal preparations and health foods by the liquid chromatography tandem with quadrupole mass spectrometry method[J].Chin Pharm Aff,2011,25(1):70-72.
[15]朱峰,阮丽萍,马永建,等.超高效液相色谱-串联质谱联用法同时检测降糖类和减肥类保健品中20种非法添加的化学降糖药物[J].色谱,2014,32(1):13-20.Zhu F,Ruan LP,Ma YJ,et al.Simultaneous detecmination of 20 illeyally added anti-diabetic chemical components in hypoglycemic and weight-reducing health foods by ultra-high performance liquid chromatography tandem mass spectrometry[J].Chin J Chromatogr,2014,32(1):13-20.