目的:采用化学分析及基因检测等技术建立风寒咳嗽丸中水稻源性成分的检测方法,制定相应的新标准,以规范药品生产企业的非法行为,保障用药安全。方法:采用高效液相色谱法对不同来源的风寒咳嗽丸中的成分进行比对,并对其淀粉含量进行测定;采用PCR方法对不同来源的样品进行基因筛查,并优化样品前处理方法和DNA提取过程,对方法的检出限、精密度和重复性进行考察。结果:不同来源的样品指纹图谱明显不同,淀粉含量差异很大。PCR方法证明了问题样品掺入了水稻源性成分,该方法检测限为1 g·kg-1,精密度及重复性良好;实验结果表明2批次问题样品和1批次问题混合粉均掺入了水稻源性成分。结论:建立了风寒咳嗽丸中掺伪水稻源性成分的PCR检验检测标准,弥补了此类掺伪手段在监管中的漏洞。
Objective: In order to regulate the illegal behavior of drug manufacturers and ensure the safety of drug use, chemical analysis and gene detection techniques were used to establish the detection method of rice components in Fenghan Kesou pills, and the corresponding new standards were established. Methods: The composition of Fenghan Kesou pills from different sources was compared by HPLC, and the content of starch was determined. PCR method was used to screen the genes of samples from different sources, and the sample pre-treatment method and DNA extraction process were optimized. The detection limit, precision and repeatability of the method were investigated. Results: The fingerprints of samples from different sources were obviously different, and the starch content varied a lot. PCR method proved that the samples were adulterated with rice-derived ingredients, and the detection limit of this method was 1 g·kg-1, with good precision and repeatability. The experimental results showed that the two batches of suspicious samples and one batch of suspicious mixed powder were mixed with rice-derived ingredients. Conclusion: The PCR detection standard of adulterated rice-derived ingredients in Fenghan Kesou pills is established to make up the loopholes in supervision of such adulterated means.
[1] 中国食品药品检定研究院. 国家药品抽检年报(2016)[EB/OL]. (2017-10-09)https://www.nifdc. org. cn//nifdc/bshff/gjchj/20171009132201938.html
National Institutes for Food and Drug Control. Annual Report of National Drug Sampling Test. (2016)[EB/OL]. (2017-10-09)https://www.nifdc. org. cn//nifdc/bshff/gjchj/201710091 32201938.html
[2] 中国食品药品检定研究院. 国家药品抽检年报(2017)[EB/OL]. (2018-06-05)https://www.nifdc. org. cn//nifdc/bshff/gjchj/20180605132401655.html
National Institutes for Food and Drug Control. Annual Report of National Drug Sampling Test. (2017)[EB/OL]. (2018-06-05)https://www.nifdc. org. cn//nifdc/bshff/gjchj/201806051 32401655.html
[3] 中国食品药品检定研究院. 国家药品抽检年报(2018)[EB/OL]. (2019-08-07) https://www.nifdc. org. cn//nifdc/bshff/gjchj/20190807102557426.html
National Institutes for Food and Drug Control. Annual Report of National Drug Sampling Test. (2018)[EB/OL]. (2019-08-07) https://www.nifdc.org.cn//nifdc/bshff/gjchj/20190807102557426.html
[4] 中国食品药品检定研究院. 国家药品抽检年报(2019)[EB/OL]. (2020-03-23) https://www.nifdc.org.cn//nifdc/bshff/gjchj/20200323143900758.html
National Institutes for Food and Drug Control. Annual Report of National Drug Sampling Test. (2019)[EB/OL]. (2020-03-23) https://www.nifdc.org.cn//nifdc/bshff/gjchj/20200323143900758.html
[5] 中国食品药品检定研究院. 国家药品抽检年报(2020)[EB/OL]. (2021-03-19) https://www.nifdc.org.cn//nifdc/bshff/gjchj/gjchjtzgg/202103231108143186. htmL
National Institutes for Food and Drug Control. Annual Report of National Drug Sampling Test. (2020)[EB/OL]. (2021-03-19) https://www.nifdc.org.cn//nifdc/bshff/gjchj/gjchjtzgg/202103231108143186.html
[6] 中国食品药品检定研究院. 国家药品抽检年报(2021)[EB/OL]. (2022-03-18) https://www.nifdc.org.cn//nifdc/bshff/gjchj/gjchjtzgg/2022031815002228792.html
National Institutes for Food and Drug Control. Annual Report of National Drug Sampling Test. (2021)[EB/OL]. (2022-03-18) https://www.nifdc.org.cn//nifdc/bshff/gjchj/gjchjtzgg/2022031815002228792.html
[7] 刘文,王翀,朱炯. 国家药品抽检中补充检验方法和检验项目的应用现状与思考[J]. 药物评价研究,2020,43(10):1939
LIU W, WANG C, ZHU J. Current situation and review on application of supplementary testing methods and items in national drug sampling and testing[J]. Drug Eval Res, 2020, 43(10):1939
[8] 程显隆,李明华,郭晓晗,等.中药补充检验方法研究及在市场监管中的作用[J]. 中国食品药品监管,2022(3):74
CHENG XL, LI MH, GUO XH, et al. Study of supplementary testing methods for traditional chinese medicine and their application in market supervision[J]. China Food Drug Admin, 2022(3):74
[9] 马双成,王莹,魏锋.中药质量控制未来发展方向的思考[J]. 中国药学杂志,2021,56(16):1273
MA SC, WANG Y, WEI F. Ideas on future development direction of quality control of traditional Chinese medicine[J]. Chin Pharm J, 2021, 56(16):1273
[10] 何轶,李静,余坤子,等.某抗风湿中药医院制剂中添加结晶样物质的检测[J]. 中国药学杂志,2020,55(7):549
HE Y, LI J, YU KZ, et al. Analysis of crystalline substances in a TCM hospital preparation for treatment of Rheumatism[J]. Chin Pharm J, 2020, 55(7):549
[11] 王菲菲,任秀,李静,等. 关于中成药水蜜丸制剂掺伪米粉监管方法的研究[J]. 中国药事,2021,35(12):1364
WANG FF, REN X, LI J, et al. Research on the supervision method about the detection of adulterated rice in Chinses patent medicines[J]. Chin Pharm Aff, 2021, 35(12):1364
[12] 中华人民共和国药典2020年版.一部[S]. 2020:727
ChP 2020. Vol Ⅰ [S]. 2020:727
[13] 王峰,何轶,于健东,等.牛黄清胃丸中麦冬掺伪情况研究[J]. 中国药学杂志,2019,54(16):1332
WANG F, HE Y, YU JD, et al. Adulteration situation of Ophiopogonis Radix in Niuhuang Qingwei pills[J]. Chin Pharm J, 2019, 54(16):1332
[14] 付凌燕,闵春艳,汪祺,等.市售西红花药材掺伪染色检测方法的实验研究[J]. 药物分析杂志,2012,32(01):74
FU LY, MIN CY, WANG Q, et al. Experimental research on the detection methods of Croci Stigma adulteration on the markets[J]. Chin J Pharm Anal, 2012, 32(1):74
[15] 聂黎行,张烨,张南平,等.显微技术在牛黄清心丸(局方)质量控制中的应用[J]. 中国中药杂志,2016,41(20):3741
NIE LX, ZHANG Y, ZHANG NP, et al. Application of microscopic spectroscopy in quality control of Niuhuang Qingxin pills[J]. China J Chin Mater Med, 2016, 41(20):3741
[16] 闫建功,王亚丹,戴忠,等. 酶解-HPLC-ELSD法测定防风通圣丸中淀粉含量[J]. 中国食品药品监管,2021,212(9):122
YAN JG, WANG YD, DAI Z, et al. Determination of starch in Fangfeng Tongsheng pills by enzymatic hydrolysis-HPLC-ELSD method[J]. China Food Drug Admin, 2021, 212(09):122
[17] 王菲菲,任秀,李静,等.九味羌活丸和香砂养胃丸制剂米粉掺伪检测方法的建立和质量标准的制定[J]. 暨南大学学报(自然科学与医学版),2022,43(1):97
WANG FF, REN X, LI J, et al. Establishment of quality standards and test method of rice adulteration in Jiuweiqianghuo Wan and Xiangshayangwei Wan[J]. J Jinan Univ (Nat Sci Med), 2022, 43(1):97
[18] 刘晶晶,王菲菲,任秀,等.基于特异性聚合酶链式反应方法鉴别九味羌活丸中水稻源性成分掺伪[J]. 中国药学杂志,2021,56(23):1894
LIU JJ, WANG FF, REN X, et al. Identification of adulteration of rice-derived ingredients in Jiuwei Qianghuo pills with specific PCR method [J]. Chin Pharm J, 2021, 56(23):1894
[19] 王菲菲,刘杰,何风艳,等.薄荷属3种植物及变种的性状鉴别、化学成分分析及DNA条形码研究比较[J]. 药学学报,2019,54(11):2083
WANG FF, LIU J, HE FY, et al. Comparative study for morphological identification, chemical component analysis, and DNA barcode determination of three plants and their varieties in Genus Mentha[J]. Acta Pharm Sin, 2019, 54(11):2083
