高效液相色谱-串联质谱法测定盐酸二甲双胍及其制剂中痕量<i>N</i>-亚硝基二甲胺

引用本文: 郭常川, 刘琦, 张雷, 郑静, 汪勇, 杨书娟, 褚志杰, 牛冲, 徐玉文. 高效液相色谱-串联质谱法测定盐酸二甲双胍及其制剂中痕量N-亚硝基二甲胺[J]. 色谱, 2020, 38(11): 1288-1293. doi: 10.3724/SP.J.1123.2020.03008 shu

Citation: GUO Changchuan, LIU Qi, ZHANG Lei, ZHENG Jing, WANG Yong, YANG Shujuan, CHU Zhijie, NIU Chong, XU Yuwen. Determination of N-nitrosodimethylamine in metformin hydrochloride and its preparations by high performance liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Chromatography, 2020, 38(11): 1288-1293. doi: 10.3724/SP.J.1123.2020.03008 shu

高效液相色谱-串联质谱法测定盐酸二甲双胍及其制剂中痕量N-亚硝基二甲胺

郭常川 ^1, ,
刘琦 ^1, ,
张雷 ^1, ,
郑静 ^1, ,
汪勇 ^2, ,
杨书娟 ^1, ,
褚志杰 ^3, ,
牛冲 ^1, ,
徐玉文 ^{1,
,}

1.
山东省食品药品检验研究院, 山东济南 250101;
2.
岛津企业管理(中国)有限公司, 北京 100020;
3.
山东省中医院, 山东济南 250014

通讯作者: 徐玉文, E-mail:xuyuwen250101@126.com

基金项目:
国家自然科学基金项目（81573606，81973486）.

摘要: 建立了高效液相色谱-串联质谱（HPLC-MS/MS）检测盐酸二甲双胍原料和制剂中N-亚硝基二甲胺（NDMA）含量的方法。样品以水为提取溶剂，经涡旋混匀、恒温振荡、高速离心、微孔过滤后进行HPLC-MS/MS分析。采用ACE EXCEL 3 C18-AR色谱柱（150 mm×4.6 mm，3 μm）分离，流动相为均含0.1%甲酸的水和甲醇溶液，梯度洗脱，流速0.8 mL/min，柱温40℃，自动进样器温度10℃。采用阀切换技术保护质谱仪，设置六通阀切换使保留时间2.85~7.00 min的流动相进入质谱，其余时间流动相进入废液。质谱部分采用大气压化学电离（APCI）源，在正离子、MRM模式下扫描，雾化器流量为3 L/min，加热器流量为10 L/min，接口温度为300℃，脱溶剂管温度为250℃，加热块温度为400℃，干燥器流量为10 L/min。NDMA定量离子对为m/z 75.0→43.1，碰撞能量（CE）为-17.0 eV，定性离子对为m/z 75.0→58.2，CE为-16.0 eV。采用外标法定量。对方法进行了详细的方法学验证，结果表明，该法专属性良好，溶剂和辅料对NDMA测定无干扰。NDMA峰面积与其质量浓度在1.00~100.00 ng/mL范围内呈现良好的线性关系，相关系数（r）>0.9999；低、中、高3个水平下NDMA的回收率为94.55%~114.67%，RSD为4.73%~13.46%；检出限和定量限分别为0.20 ng/mL和1.00 ng/mL；NDMA在自动进样器放置0、8、24 h的峰面积RSD为2.08%。使用该方法对113批盐酸二甲双胍原料和制剂供试品中的NDMA进行测定，发现原料药中NDMA检出量不超限，但有8批二甲双胍制剂超过了限度。该法灵敏、准确，操作简便，可用于盐酸二甲双胍原料及制剂中的NDMA检测。

关键词:

English

Determination of N-nitrosodimethylamine in metformin hydrochloride and its preparations by high performance liquid chromatography-tandem mass spectrometry

1.
Shandong Institute for Food and Drug Control, Jinan 250101, China;
2.
Shimadzu(China) Co. Ltd., Beijing 100020, China;
3.
Shandong Provincial Hospital of Traditional Chinese Medicine, Jinan 250014, China

Corresponding author: XU Yuwen, xuyuwen250101@126.com

Received Date: 12 March 2020
Fund Project:
National Natural Science Foundation of China (Nos. 81573606, 81973486).

Abstract: A method was established for the determination of N-nitrosodimethylamine (NDMA) in metformin hydrochloride active pharmaceutical ingredient (API) and preparation samples by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Water was used as the extraction solvent for the metformin hydrochloride API and preparation samples. The samples were analyzed by HPLC-MS/MS after vortex mixing, constant temperature shaking, high speed centrifugation and microfiltration. An ACE EXCEL 3 C18-AR column (150 mm×4.6 mm, 3 μm) was used for chromatographic separation. The mobile phases were water and methanol both containing 0.1% formic acid with gradient elution. The flow rate, column temperature, and autosampler temperature were set as 0.8 mL/min, 40℃, and 10℃, respectively. The valve switching technique was used to protect the mass spectrometer, while six-way valve switching was adopted to allow the mobile phase with a retention time of 2.85-7.00 min to enter the mass spectrometer and the mobile phase with other retention times to enter the waste liquid. For the mass spectrometer, an atmospheric pressure chemical ionization (APCI) ion source was used in positive ion MRM scanning mode. The other conditions were as follows:atomizer flow, 3 L/min; heater flow, 10 L/min, interface temperature, 300℃; desolvation line (DL) temperature, 250℃; heating block temperature, 400℃; and dryer flow, 10 L/min. The quantitative transition of NDMA was m/z 75.0→43.1 with a collision energy of-17.0 eV, while the qualitative transition was m/z 75.0→58.2 with a collision energy of-16.0 eV. The external standard method was utilized for quantitative analysis. The established method was validated in detail by investigating the specificity, linear range, limit of detection, limit of quantification, recovery, precision, and stability. This method showed good specificity, since the solvents and excipients did not interfere with the determination of NDMA. A good linear relationship was observed the NDMA peak area and the mass concentrations in the range of 1.00-100.00 ng/mL with an excellent correlation coefficient (r>0.9999). The limit of detection and limit of quantification in solution were 0.20 ng/mL and 1.00 ng/mL, respectively. The recoveries of NDMA at low, medium, and high spiked levels ranged from 94.55% to 114.67%, and the RSDs ranged from 4.73% to 13.46%, indicating good accuracy and precision for the quantification of NDMA. Stability tests showed that NDMA was stable when placed in the autosampler for 0, 8, 24 h, since the RSD of the peak area was as low as 2.08%. The validated method was then applied to the determination of NDMA in metformin hydrochloride raw materials and preparations (tablets, capsules or enteric tablets). The detected amount of NDMA in the API did not exceed the limit in 113 batches of samples, but NDMA was detected and exceeded the limit in eight batches of preparations. This method is sensitive, accurate, and easy to operate, and it can be used for the determination of NDMA in metformin hydrochloride raw materials and preparation samples.

Key words:

high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS)
/ trace determination
/ N-nitrosodimethylamine (NDMA)
/ metformin

1. [1] International Council for Harmonization (ICH). ICH Harmonised Guideline, Assessment and Control of DNA Reactive (Mutagenic) Impurities in Pharmaceuticals to Limit Potential Carcinogenic Risk M7 (R1). (2017-03-31). https://www.ema.europa.eu/en/documents/scientific-guideline/guideline-assessment-control-dna-reactive-mutagenic-impurities-pharmaceuticals-limit-potential_en.pdf
2. [2] International Agency for Research on Cancer (IARC). Agents Classified by the IARC Monographs.[2020-03-03]. http://monographs.iarc.fr/ENG/Classification/index.php
3. [3] Lin C H, Wang P H, Wang T H, et al. Nanoscale, 2020, 12(2):1075
4. [4] Ceto X, Saint C P, Chow C W K, et al. Sens Actuators B:Chem, 2016, 237:613
5. [5] Kodamatani H, Yamasaki H, Sakaguchi T, et al. J Chromatogr A, 2016, 1460:202
6. [6] Zhang W W, Zhao C H, Fu M, et al. Physical Testing and Chemical Analysis (Part B:Chemical Analysis), 2019, 55 (9):1028 张伟伟, 赵春华, 付萌, 等. 理化检验(化学分册), 2019, 55(9):1028
7. [7] Wu C H, Du G F, Li T, et al. Journal of Food Safety and Quality, 2018, 9(7):1695 吴翠华, 杜高发, 李婷, 等. 食品安全质量检测学报, 2018, 9(7):1695
8. [8] GB5009.26-2016
9. [9] Choi N R, Kim Y P, Ji W H, et al. Talanta, 2016, 148:69
10. [10] Liu H, Zhang Y C, Lu Y F, et al. Physical Testing and Chemical Analysis (Part B:Chemical Analysis), 2017, 53(8):917 刘鸿, 张怡春, 陆怡峰, 等. 理化检验(化学分册), 2017, 53(8):917
11. [11] Zhang F Y, Jiang W F, Wang B, et al. Chinese Journal of Health Laboratory Technology, 2018, 28(11):1306 张凤艳, 蒋万枫, 王蓓, 等. 中国卫生检验杂志, 2018, 28(11):1306
12. [12] Li D K, Zhang Y, Liu X P, et al. Chinese Journal of Chromatography, 2019, 37(2):216 李登昆, 张云, 刘祥萍, 等. 色谱, 2019, 37(2):216
13. [13] Zhu M M, Ye Q, Zhou T T, et al. Chinese Journal of Chromatography, 2019, 37(2):207 朱萌萌, 叶群, 周婷婷, 等. 色谱, 2019, 37(2):207
14. [14] Jiang J, Zou Y, Sun L P. Chinese Journal of Analysis Laboratory, 2020, 39(2):245 姜俊, 邹韵, 孙丽鹏. 分析试验室, 2020, 39(2):245
15. [15] Wu Z W, Du K, Wang L, et al. Chinese Journal of New Drugs, 2019, 28(20):2478 吴兆伟, 杜凯, 王琳, 等. 中国新药杂志, 2019, 28(20):2478
16. [16] Xian R Q, Gong L P, Xing S, et al. Chinese Journal of Pharmaceutical Analysis, 2019, 39(8):1501 咸瑞卿, 巩丽萍, 邢晟, 等. 药物分析杂志, 2019, 39(8):1501
17. [17] Sun C Y, Ji Y H, Qin K M, et al. Chinese Journal of Chromatography, 2019, 37(12):1297 孙春艳, 纪颖鹤, 秦昆明, 等. 色谱, 2019, 37(12):1297
18. [18] Zhou Y F, Wang F H, Wang Z L, et al. Chinese Journal of Chromatography, 2018, 36(2):159 周艳芬, 王芳焕, 王泽岚, 等. 色谱, 2018, 36(2):159
19. [19] National Medical Products Administration. Technical Guidelines for Research on Nitrosamine Impurities in Chemical Drugs (for Trial Implementation). (2020-05-08). http://www.nmpa.gov.cn/WS04/CL2138/377043.html 国家药品监督管理局. 化学药物中亚硝胺类杂质研究技术指导原则(试行). (2020-05-08). http://www.nmpa.gov.cn/WS04/CL2138/377043.html

扫一扫看文章

计量

PDF下载量: 2
文章访问数: 72
HTML全文浏览量: 6

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

高效液相色谱-串联质谱法测定盐酸二甲双胍及其制剂中痕量N-亚硝基二甲胺

通讯作者: 徐玉文, E-mail:xuyuwen250101@126.com

English

Determination of N-nitrosodimethylamine in metformin hydrochloride and its preparations by high performance liquid chromatography-tandem mass spectrometry

Corresponding author: XU Yuwen, xuyuwen250101@126.com

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

高效液相色谱-串联质谱法测定盐酸二甲双胍及其制剂中痕量N-亚硝基二甲胺

通讯作者: 徐玉文, E-mail:xuyuwen250101@126.com

English

Determination of N-nitrosodimethylamine in metformin hydrochloride and its preparations by high performance liquid chromatography-tandem mass spectrometry

Corresponding author: XU Yuwen, xuyuwen250101@126.com

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs