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
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摘要
目的建立高效液相色谱法测定具有调节血糖功能袋泡茶中4种非法添加化学药物(二甲双胍、格列吡嗪、格列本脲、格列美脲)的含量的分析方法。方法采集7种调节血糖功能袋泡茶类保健食品,经甲醇提取,过0.22μm滤膜上机。采用Shim-Pack VP-ODS色谱柱(4.6 mm×250 mm,4.6μm);以甲醇(A),0.02 mol/L磷酸二氢铵水溶液(B)为流动相进行梯度洗脱,流速为1.0 m L/min,柱温25℃,外标法定量。结果二甲双胍、格列吡嗪、格列本脲、格列美脲检出限分别为0.033、0.040、0.041、0.037 mg/L,加标回收率为93.74%～104.31%,相对标准偏差均低于4.99%。7件样品中有1件检测出含格列吡嗪,其余降低血糖作用化学药物成分未检出。结论市面上存在添加化学药物成分的调节血糖功能袋泡茶类保健食品,应加强保健食品的监管。
Objective To establish a method for simultaneous determination the content of 4 illegal added chemicals(metformin, glipizide, glibenclamide and glimepiride) in teabags with blood sugar regulating function by high performance liquid chromatography. Methods Seven kinds of health foods of teabags with regulating blood sugar function were collected and extracted by methanol, then the supernatant was filtered through 0.22 μm membrane for analysis. The samples were separated with a Shim-Pack VP-ODS column(4.6 mm×250 mm, 4.6 μm) at the flow rate of 1.0 m L/min by gradient elution using methanol(solvent A) and 0.02 mol/L ammonium dihydrogen phosphate solution(solvent B), the temperature of the column was maintained at 25 ℃, and quantitated by external standard method. Results The limits of detection of metformin, glipizide, glibenclamide and glimetazide were 0.033, 0.040, 0.041 and 0.037 mg/L, respectively, the recoveries were 93.74%-104.31%, and RSD were less than 4.99%. Glipizide was detected in one of the seven samples, while the rest of the chemical ingredients for hypoglycemic drugs were not detected. Conclusion Partial teabags with the function of regulating blood sugar in the market add chemical drugs, so the supervision of health food should be strengthened.
Objective To establish a method for simultaneous determination the content of 4 illegal added chemicals(metformin, glipizide, glibenclamide and glimepiride) in teabags with blood sugar regulating function by high performance liquid chromatography. Methods Seven kinds of health foods of teabags with regulating blood sugar function were collected and extracted by methanol, then the supernatant was filtered through 0.22 μm membrane for analysis. The samples were separated with a Shim-Pack VP-ODS column(4.6 mm×250 mm, 4.6 μm) at the flow rate of 1.0 m L/min by gradient elution using methanol(solvent A) and 0.02 mol/L ammonium dihydrogen phosphate solution(solvent B), the temperature of the column was maintained at 25 ℃, and quantitated by external standard method. Results The limits of detection of metformin, glipizide, glibenclamide and glimetazide were 0.033, 0.040, 0.041 and 0.037 mg/L, respectively, the recoveries were 93.74%-104.31%, and RSD were less than 4.99%. Glipizide was detected in one of the seven samples, while the rest of the chemical ingredients for hypoglycemic drugs were not detected. Conclusion Partial teabags with the function of regulating blood sugar in the market add chemical drugs, so the supervision of health food should be strengthened.
引文
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[6]梁燕,王岳飞,谢争珍,等.茶桑混合袋泡茶降血糖作用的实验研究[J].茶叶科学,2008,(5):358-362.Liang Y,Wang YF,Xie ZZ,et al.Research on the effect of tea and mulberry mixture tea bag on blood glucose-decrease in diabetic model mice[J].J Tea Sci,2008,(5):358-362.
[7]王东波,唐华茂,葛献英,等.揭秘“三品一械”市场“十大”骗术[J].首都医药,2013,20(13):19-20.Wang DB,Tang HM,Ge XY,et al.The"top ten"scam in the"three pinkies"market[J].Capit Med,2013,20(13):19-20.
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[9]王俊苏,郗存显,陈冬东,等.HPLC法检测降血糖保健食品中的双胍类药物[J].中国药房,2013,24(1):68-71.Wang JS,Xi CX,Chen DD,et al.Determination of biguanides in anti-diabetic health foods by HPLC[J].Chin Pharm,2013,24(1):68-71.
[10]张振华,陶妙娟.四种中成药及保健品非法添加化学药物的快速筛查方法初探[J].安徽医药,2008,12(1):28-29.Zhang ZH,Tao MJ.A quick screening method for four kinds of Chinese patent medicines and health products illegally added chemical drugs[J].Anhui Med Pharm J,2008,12(1):28-29.
[11]田雁钰.参芪降糖颗粒中非法添加格列本脲的检测[J].中国药房,2008,19(27):2131-2132.Tian YY.Detection of glibenclamide mixed illegally in shenqi hypoglycemic granules[J].Chin Pharm,2008,19(27):2131-2132.
[12]刘福艳,谢元超,刘福强,等.液相色谱—质谱联用法检测2种中药降糖制剂中非法掺入的二甲双胍、苯乙双胍、格列吡嗪、格列本脲[J].药物分析杂志,2009,29(3):440-444.Liu FY,Xie YC,Liu FQ,et al.LC-MS determination of metformin,phenformin,glipizide and glyburide illegally mixed into two kind of antidiabetic preparations of traditional Chinese medicine[J].Chin J Pharm Anal,2009,29(3):440-444.
[13]夏瑞,张喆.液相色谱-质谱方法鉴别中药降糖药中的格列吡嗪[J].分析测试学报,2005,24(6):28.Xia R,Hang J.Identification of glipizide in chinese traditional medicine used to cure diabetes by HPLC–MS[J].J Instrum Anal,2005,24(6):28.
[14]姜建国,张西如.应用LC-MS技术检测降糖保健品中非法添加的格列美脲[J].中国实用医药,2008,(18):67-68.Jiang JG,Zhang XR.Detection of glimepiride mixed into drug by LC-MS[J].China Pract Med,2008,(18):67-68.
[15]张昊天,孙超,宋志铭.近红外聚类分析法快速检验中成药及保健品中的非法添加4种降糖类药物[J].黑龙江医药,2017,30(2):237-241.Zhang HT,Sun C,Song ZM.Rapid detection of 4 kinds of hypoglycemic drugs illegally added in Chinese patent medicine and health care products by near infrared cluster analysis[J].Heilongjiang Med Pharm,2017,30(2):237-241.
[16]施亚琴,姚静,张启明,等.薄层色谱法快速筛查降血糖中成药及保健食品中添加磺酰脲类化学降糖药[J].药物分析杂志,2007,27(1):36-39.Shi YQ,Yao J,Zhang QM,et al.TLC method for screening sulphonylureas in antidiabetic Chinese traditional patent medicines and health supplementary foods[J].Chin J Pharm Anal,2007,27(1):36-39.
[17]李亚琴,钱丽华,申兰慧.薄层色谱法快检降糖中药添加的格列齐特[J].药物分析杂志,2008,28(2):319-320.Li YQ,Qian LH,Shen LH.TLC for detection of geliqite added in antidiabetics traditional Chinese medicine[J].Chin J Pharm Anal,2008,28(2):319-320.
[18]Starek M,Krzek J,Tarsa M,et al.Determination of piroxicam and deg radation products in drugs by TLC[J].Chromatographia,2009,69(1):351-356.
[19]Ekiert RJ,Krzek J,Wtodzimierz R,et al.Evaluation of a TLC densito metric method for analysis of azole antifungal agents[J].Chromatographia,2008,67(3):995-998.
[20]张毅.济南市售降糖保健品质量调查及西药成分检测[D].济南:山东大学,2015.Zhang Y.Research into quality levels of hypoglycemic products in Jinan,and the content of adulterated western drugs in them[D].Jinan:Shandong University,2015.
[21]袁向华,鲁玉侠,蔡妙颜,等.蜂胶中盐酸二甲双胍、盐酸苯乙双胍和格列本脲HPLC测定方法的建立[J].食品研究与开发,2007,2(7):125-127.Yuan XH,Lv YX,Cai MY,et al.The foundation of mensuration method of metformin hydrochloride,phenformin hydrochloride and glibenclamid in propolis by HPLC[J].Food Res Dev,2007,2(7):125-127.
[22]郭栋,那顺朝克图,王江华,等.高效液相色谱法测定降糖中成药中添加的高极性化学降糖药物[J].色谱,2009,27(2):211-215.Guo D,Nashunchaoketu,Wang JH,et al.Simultaneous determination of four highly polar anti-diabetic drugs in Chinese traditional patent medicines using high performance liquid chromatography[J].Chin J Chromatogr,2009,27(2):211-215.
[23]郑文燕,李哲,陈梅,等.LC-MS/MS法检测中成药及保健品中减肥和降糖类的非法添加药物[J].沈阳药科大学学报,2018,35(1):58-66,82.Zheng WY,Li Z,Chen M,et al.Determination of anti-obesity and anti-diabetic chemicals illegally added into traditional Chinese medicines and health care products by LC-MS/MS[J].J Shenyang Pharm Univ,2018,35(1):58-66,82.
[24]郭继芬,陈笑艳,钟大放.6种口服降糖药的液相色谱-质谱分析[J].分析测试学报,2000,19(6):5-8.Guo JF,Chen XY,Zhong DF.Analysis of antidiabetics by liquid chromatography-mass spectrometry[J].J Instrum Anal,2000,19(6):5-8.
[25]董宇,孔璋,钟大放.液相色谱-质谱联用法检测中药降糖制剂中非法掺入的苯乙双胍和格列本脲[J].沈阳药科大学学报,2005,22(1):19-22.Dong Y,Kong Z,Zhong DF.Detection of phenformin and glyburide illegally mixed into traditional Chinese medicine antidiabetic preparations by the liquid chromatography-mass spectrometry method[J].J Shenyang Pharm Univ,2005,22(1):19-22.
[26]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):862-864.Luo 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):862-864.
[27]Patil P,Vaidya I.Overview on impurity profiling[J].Int J Pharm Res Sch,2013,2(2):54-65.
[28]Ruhland M,Tischler J.Determination of ergot alkaloids in feed by HPLC[J].Mycotoxin Res,2008,24(2):73-79.
[29]Kakadiya PR,Chandrashekhar TG,Ganguly S,et al.Low level determinations of methyl methanesulfonate and ethyl methanesulfonate impurities in emtricitabine active pharmaceutical Ingredient by LC/MS/MS using electrospray ionization[J].Anal Chem,2011,6:1-28.
[30]杨春平,李钊君,宁红,等.HPLC检测枇杷中氯吡脲含量的前处理方法[J].浙江大学学报(农业与生命科学版),2016,42(3):350-357.Yang CP,Li ZJ,Ning H,et al.Pretreatment method of forchlorfenuron residues in loquat by HPLC analysis[J].J Zhejiang Univ(Agric Life Sci),2016,42(3):350-357.
[2]Buse JB,Ginsberg HN,Bakris GL,et al.Primary prevention of cardiovascular diseases in people with diabetes mellitus:A scientific statement from the American Heart Association and the American Diabetes Association[J].Diabetes Care,2007,30(1):162-172.
[3]Hong X,Chen X,Chu J,et al.Multiple diabetic complications,as well as impaired physical and mental function,are associated with declining balance function in older persons with diabetes mellitus[J].Clin Interv Aging,2017,12:189-195.
[4]江秀慧.老年糖尿病并发症的临床分析[J].中国实用医药,2015,10(19):86-87.Jiang XH.Clinical analysis of complications of diabetes in the elderly[J].Chin Prac Med,2015,10(19):86-87.
[5]傅樱花,张富春.降血糖功能性食品的研究现状[J].食品研究与开发,2011,32(11):165-168.Fu YH,Zhang FC.Status of study on functional food with antidiabetic activity[J].Food Res Dev,2011,32(11):165-168.
[6]梁燕,王岳飞,谢争珍,等.茶桑混合袋泡茶降血糖作用的实验研究[J].茶叶科学,2008,(5):358-362.Liang Y,Wang YF,Xie ZZ,et al.Research on the effect of tea and mulberry mixture tea bag on blood glucose-decrease in diabetic model mice[J].J Tea Sci,2008,(5):358-362.
[7]王东波,唐华茂,葛献英,等.揭秘“三品一械”市场“十大”骗术[J].首都医药,2013,20(13):19-20.Wang DB,Tang HM,Ge XY,et al.The"top ten"scam in the"three pinkies"market[J].Capit Med,2013,20(13):19-20.
[8]国家食品药品监督管理局.保健食品中可能非法添加的物质[EB/OL].[2013-01-14].http://www.sfda.gov.cn/WS01/CL1162/76614.html.State Food and Drug Administration.Substances that may be illegally added to health food[EB/OL].[2013-01-14].http://www.sfda.gov.cn/WS01/CL1162/76614.html.
[9]王俊苏,郗存显,陈冬东,等.HPLC法检测降血糖保健食品中的双胍类药物[J].中国药房,2013,24(1):68-71.Wang JS,Xi CX,Chen DD,et al.Determination of biguanides in anti-diabetic health foods by HPLC[J].Chin Pharm,2013,24(1):68-71.
[10]张振华,陶妙娟.四种中成药及保健品非法添加化学药物的快速筛查方法初探[J].安徽医药,2008,12(1):28-29.Zhang ZH,Tao MJ.A quick screening method for four kinds of Chinese patent medicines and health products illegally added chemical drugs[J].Anhui Med Pharm J,2008,12(1):28-29.
[11]田雁钰.参芪降糖颗粒中非法添加格列本脲的检测[J].中国药房,2008,19(27):2131-2132.Tian YY.Detection of glibenclamide mixed illegally in shenqi hypoglycemic granules[J].Chin Pharm,2008,19(27):2131-2132.
[12]刘福艳,谢元超,刘福强,等.液相色谱—质谱联用法检测2种中药降糖制剂中非法掺入的二甲双胍、苯乙双胍、格列吡嗪、格列本脲[J].药物分析杂志,2009,29(3):440-444.Liu FY,Xie YC,Liu FQ,et al.LC-MS determination of metformin,phenformin,glipizide and glyburide illegally mixed into two kind of antidiabetic preparations of traditional Chinese medicine[J].Chin J Pharm Anal,2009,29(3):440-444.
[13]夏瑞,张喆.液相色谱-质谱方法鉴别中药降糖药中的格列吡嗪[J].分析测试学报,2005,24(6):28.Xia R,Hang J.Identification of glipizide in chinese traditional medicine used to cure diabetes by HPLC–MS[J].J Instrum Anal,2005,24(6):28.
[14]姜建国,张西如.应用LC-MS技术检测降糖保健品中非法添加的格列美脲[J].中国实用医药,2008,(18):67-68.Jiang JG,Zhang XR.Detection of glimepiride mixed into drug by LC-MS[J].China Pract Med,2008,(18):67-68.
[15]张昊天,孙超,宋志铭.近红外聚类分析法快速检验中成药及保健品中的非法添加4种降糖类药物[J].黑龙江医药,2017,30(2):237-241.Zhang HT,Sun C,Song ZM.Rapid detection of 4 kinds of hypoglycemic drugs illegally added in Chinese patent medicine and health care products by near infrared cluster analysis[J].Heilongjiang Med Pharm,2017,30(2):237-241.
[16]施亚琴,姚静,张启明,等.薄层色谱法快速筛查降血糖中成药及保健食品中添加磺酰脲类化学降糖药[J].药物分析杂志,2007,27(1):36-39.Shi YQ,Yao J,Zhang QM,et al.TLC method for screening sulphonylureas in antidiabetic Chinese traditional patent medicines and health supplementary foods[J].Chin J Pharm Anal,2007,27(1):36-39.
[17]李亚琴,钱丽华,申兰慧.薄层色谱法快检降糖中药添加的格列齐特[J].药物分析杂志,2008,28(2):319-320.Li YQ,Qian LH,Shen LH.TLC for detection of geliqite added in antidiabetics traditional Chinese medicine[J].Chin J Pharm Anal,2008,28(2):319-320.
[18]Starek M,Krzek J,Tarsa M,et al.Determination of piroxicam and deg radation products in drugs by TLC[J].Chromatographia,2009,69(1):351-356.
[19]Ekiert RJ,Krzek J,Wtodzimierz R,et al.Evaluation of a TLC densito metric method for analysis of azole antifungal agents[J].Chromatographia,2008,67(3):995-998.
[20]张毅.济南市售降糖保健品质量调查及西药成分检测[D].济南:山东大学,2015.Zhang Y.Research into quality levels of hypoglycemic products in Jinan,and the content of adulterated western drugs in them[D].Jinan:Shandong University,2015.
[21]袁向华,鲁玉侠,蔡妙颜,等.蜂胶中盐酸二甲双胍、盐酸苯乙双胍和格列本脲HPLC测定方法的建立[J].食品研究与开发,2007,2(7):125-127.Yuan XH,Lv YX,Cai MY,et al.The foundation of mensuration method of metformin hydrochloride,phenformin hydrochloride and glibenclamid in propolis by HPLC[J].Food Res Dev,2007,2(7):125-127.
[22]郭栋,那顺朝克图,王江华,等.高效液相色谱法测定降糖中成药中添加的高极性化学降糖药物[J].色谱,2009,27(2):211-215.Guo D,Nashunchaoketu,Wang JH,et al.Simultaneous determination of four highly polar anti-diabetic drugs in Chinese traditional patent medicines using high performance liquid chromatography[J].Chin J Chromatogr,2009,27(2):211-215.
[23]郑文燕,李哲,陈梅,等.LC-MS/MS法检测中成药及保健品中减肥和降糖类的非法添加药物[J].沈阳药科大学学报,2018,35(1):58-66,82.Zheng WY,Li Z,Chen M,et al.Determination of anti-obesity and anti-diabetic chemicals illegally added into traditional Chinese medicines and health care products by LC-MS/MS[J].J Shenyang Pharm Univ,2018,35(1):58-66,82.
[24]郭继芬,陈笑艳,钟大放.6种口服降糖药的液相色谱-质谱分析[J].分析测试学报,2000,19(6):5-8.Guo JF,Chen XY,Zhong DF.Analysis of antidiabetics by liquid chromatography-mass spectrometry[J].J Instrum Anal,2000,19(6):5-8.
[25]董宇,孔璋,钟大放.液相色谱-质谱联用法检测中药降糖制剂中非法掺入的苯乙双胍和格列本脲[J].沈阳药科大学学报,2005,22(1):19-22.Dong Y,Kong Z,Zhong DF.Detection of phenformin and glyburide illegally mixed into traditional Chinese medicine antidiabetic preparations by the liquid chromatography-mass spectrometry method[J].J Shenyang Pharm Univ,2005,22(1):19-22.
[26]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):862-864.Luo 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):862-864.
[27]Patil P,Vaidya I.Overview on impurity profiling[J].Int J Pharm Res Sch,2013,2(2):54-65.
[28]Ruhland M,Tischler J.Determination of ergot alkaloids in feed by HPLC[J].Mycotoxin Res,2008,24(2):73-79.
[29]Kakadiya PR,Chandrashekhar TG,Ganguly S,et al.Low level determinations of methyl methanesulfonate and ethyl methanesulfonate impurities in emtricitabine active pharmaceutical Ingredient by LC/MS/MS using electrospray ionization[J].Anal Chem,2011,6:1-28.
[30]杨春平,李钊君,宁红,等.HPLC检测枇杷中氯吡脲含量的前处理方法[J].浙江大学学报(农业与生命科学版),2016,42(3):350-357.Yang CP,Li ZJ,Ning H,et al.Pretreatment method of forchlorfenuron residues in loquat by HPLC analysis[J].J Zhejiang Univ(Agric Life Sci),2016,42(3):350-357.