衍生化HPLC法测定酒石酸伐尼克兰中甲醛和乙二醛的含量

管艺同; 胡芃薇; 邹文宇; 陆宇婷; 宋敏; 杭太俊

doi:10.11665/j.issn.1000-5048.20210310

衍生化HPLC法测定酒石酸伐尼克兰中甲醛和乙二醛的含量

中国药科大学药物分析系，南京 210009

基金项目: 江苏省食品药品检验监督研究院化学药品杂质谱研究重点实验室开放课题资助项目(No.NMPA-KLIPCD-2020-03)

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出版历程
- 收稿日期: 2021-01-08
- 修回日期: 2021-05-09
- 刊出日期: 2021-06-24

Determination of formaldehyde and glyoxal in varenicline tartrate using derivative method with HPLC

Department of Pharmaceutical Analysis, China Pharmaceutical University, Nanjing 210009, China

Funds: This study was supported by the Open Project of Key Laboratory of Related Substances of Chemical Medicines of Jiangsu Institute for Food and Drug Control (No.NMPA-KLIPCD-2020-03)

摘要

摘要: 建立酒石酸伐尼克兰原料药和中间体中基因毒性杂质甲醛和乙二醛的检查方法。甲醛和乙二醛经2，4-二硝基苯肼衍生化，改善HPLC-UV检查的色谱保留和检测灵敏度。利用C₈ （150 mm × 4.6 mm，5 μm）色谱柱，乙腈-水（50∶50）流动相进行线性梯度洗脱分离，在紫外光波长380 nm下进行检测，外标法测定供试品中的甲醛和乙二醛的含量。甲醛和乙二醛均在0.094 ~ 1.88 μg/L范围内线性关系良好，检测限和定量限分别达0.047 μg/mL （19 μg/g）和0.094 μg/mL （38 μg/g），平均回收率分别为（95.0 ± 1.1）%和（99.4 ± 2.6）%。经过方法学验证，本方法适用于酒石酸伐尼克兰原料药及其中间体中的微量基因毒性杂质甲醛和乙二醛的含量测定。伐尼克兰原料药及其中间体成品中甲醛和乙二醛的含量检查结果均符合每日允许最大摄入量的限度要求，而中间体粗品中乙二醛的含量易超出限度。研究所建立的方法可应用于酒石酸伐尼克兰生产工艺过程的质量控制。
- 酒石酸伐尼克兰 /
- 甲醛 /
- 乙二醛 /
- 基因毒性杂质 /
- 柱前衍生化 /
- HPLC
Abstract: To establish a method for the determination of formaldehyde and glyoxal simultaneously in varenicline tartrate active pharmaceutical ingredient (API) and its intermediate, formaldehyde and glyoxal were derivatized by 2, 4-dinitrophenylhydrazine (2,4-DNPH) to improve the HPLC retention and UV detection sensitivity. Separation was performed on a C₈ (150 mm × 4.6 mm, 5 μm) column by linear gradient elution using acetonitrile and water as the mobile phase; the detective wavelength was set at 380 nm.Formaldehyde and glyoxal were quantitatively determined by an external reference method.Linear calibration was established for both formaldehyde and glyoxal in the range from 0.094 to 1.88 μg/mL.The detection and the quantification limits were 0.047 μg/mL (19 μg/g) and 0.094 μg/mL (38 μg/g), respectively.The recoveries were (95.0±1.1)% and (99.4 ± 2.6)% for formaldehyde and glyoxal, respectively.This method has been fully validated to be applicable to quantitative analysis of trace amount of formaldehyde and glyoxal in varenicline tartrate API and its intermediate.Test results demonstrated that the contents of both formaldehyde and glyoxal met the permitted daily exposure (PDE) limits for the finished products of varenicline tartrate API as well as its intermediate, though the glyoxal contents in the crude intermediates were likely to exceed the limit.The established method is valuable for the manufacturing process and quality control of varenicline tartrate.
- varenicline tartrate /
- formaldehyde /
- glyoxal /
- genotoxic impurity /
- pre-column derivatization /
- HPLC

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参考文献(16)

[1]	. PLoS One，2020，15（4）：e0231095.
[2]	Song F，Song XN，Wang L，et al. Determination of six aniline-like genotoxic impurities in varenicline tartrate by LC-MS［J］. Chin J Pharm Anal（药物分析杂志），2018，38（1）：130-134.
[3]	Li YF，Wu XM. Synthesis and application research progress of glyoxal［J］. Fine Specialty Chem（精细与专用化学品），2020，28（3）：36-38.
[4]	Bernardini L，Barbosa E，Char?o MF，et al. Formaldehyde toxicity reports from in vitro and in vivo studies：a review and updated data［J］. Drug Chem Toxicol，2020：1-13.doi：10.1080/01480545.2020.1795190.
[5]	Barnett BM，Mu?oz ER. Effect of glyoxal pretreatment on radiation-induced genetic damage in Drosophila melanogaster［J］. Mutat Res Mol Mech Mutagen，1989，212（2）：173-179.
[6]	del Barrio MA，Hu J，Zhou PZ，et al. Simultaneous determination of formic acid and formaldehyde in pharmaceutical excipients using headspace GC/MS［J］. J Pharm Biomed Anal，2006，41（3）：738-743.
[7]	M7（R1）：Assessment and control of DNA reactive （mutagenic） impurities in pharmaceuticals to limit potential carcinogenic risk［S］：Brussels：ICH，2017：36.
[8]	EMA（European Medicines Agency）.Guidelines on the Limits of Genotoxic Impurities［S/OL］.（2006-06-28）［2019-01］.http：//www.EMA.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/09/WC500002903.pdf.
[9]	CHMP Safety Working Party.Question & Answers on the CHMP Guideline on Limits of Genotoxic Impurities［S/OL］.（2009-12）［2019-01］.http：//www.EMA.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/09/WC500002907.pdf.
[10]	Chinese Pharmacopoeia Commission.Chinese Pharmacopoeia：part 4（中华人民共和国药典：四部）［S］. Beijing：China Medical Science Press，2020.
[11]	Jiang Y，Wang J，Xu K，et al. Research on blank interference and method optimization for determination of formaldehyde and aldehyde in pharmaceutical excipients by HPLC with pre-column derivatization［J］. Chin J Pharm Anal（药物分析杂志），2020，40（11）：2062-2069.
[12]	Zhou T，Zeng S.Quantitative analysis of trace amount of formaldehyde and formic acid in hydrochlorothiazide API using ethanol derivative method with headspace gas chromatography［J］.Chin J Pharm Anal （药物分析杂志），2017，37 （5）：862-868.
[13]	Wang YP，Peng PY，Zhou YM. UV-spectrophotometric determination of glyoxylic acid and glyoxal［J］. Phys Test Chem Anal Part B Chem Anal（理化检验化学分册），2005，41 （3）：199-202.
[14]	Chen XM，Li T. Simultaneous determination of glyoxal and formaldehyde content in food paper packages［J］. Packag Food Mach（包装与食品机械），2020，38（5）：64-67.
[15]	Bicking MKL，Marcus Cooke W，Kawahara FK，et al. Effect of pH on the reaction of 2，4-dinitrophenylhydrazine with formaldehyde and acetaldehyde［J］. J Chromatogr A，1988，455：310-315.
[16]	China National Textile And Apparel Council. GB/T 2912.3-2009.Textiles-Determination of formaldehyde-Part 3：High performance liquid chromatography method（纺织品-甲醛的测定-第三部分：高效液相色谱法）［S］：Beijing：General Administration of Quality Supervision，Inspection and Quarantine of the People''s Republic of China，2009：4.