反相离子对高效液相色谱法测定左卡尼汀注射液中杂质A的含量

目的: 建立离子对高效液相色谱法测定左卡尼汀注射液中杂质A的含量。 方法: 采用Zorbax SB-C₁₈色谱柱(4.6 mm×250 mm,5 μm),流动相为磷酸盐缓冲液-甲醇(95:5),流速1.0 mL·min^-1,检测波长为215 nm。 结果: 左卡尼汀杂质A与主药峰得到良好分离,分离度达到1.772,在1~100 μg·mL ^-1的范围内,杂质A浓度与峰面积之间呈现良好线性关系,r²=0.9996(n=6),3批左卡尼汀注射液中杂质A的平均含量为0.0052%。 结论: 该方法简便、准确、专属,可用于该注射液的质量控制。

Objective: To establish a method to determine the impurity A content of levocarnitine injection by the ion pair high performance liquid chromatography. Methods: The Zorbax SB-C₁₈ column(4.6 mm×250 mm,5 μm)was used with a mobile phase consisting of phosphate buffer solution and methanol(95:5)at a flow rate of 1.0 mL·min^-1;the detection wavelength was 215 nm. Results: Levocarnitine and impurity A was completely separated by this method,and their resolution was 1.772.The linearity of peak area of impurity A was good in the range of 1-100 μg·mL^-1(r²=0.9996,n=6).The average content of impurity A in levocarnitine injection was 0.0052%. Conclusions: The established method is specific,accurate and convenient,which can be used for quality control of levocarnitine injection.

1 The National Drug Standard,New Drug Positive Standard(国家药品标准·新药转正标准).64(第64册):128
2 EP 6.0.2008.2257
3 USP32-NF27.2009.2766
4 Matsumoto K,Ichitani Y,Ogasawara N, et al.Precolumn fluorescence derivatization of eamitine and aeylcarnitines with 4-(2-aminoethylamino)-7-nitro-2,1,3-benzoxadiazole. J Chromatogr A,1994,678:241
5 De Witt P,Deias R,Muck S, et al. High-performance liquid chromatography and capillary electrophoresis of L-and D-carnitine by precolunm diastereomerie defivatization. J Chromatogr B Biomed Appl ,1994,657(1):67
6 GAO Li-jun(高立军),WANG Tao(王涛),QUAN Dong-qin(全东琴).A HPLC-method of determination of content of levocarnitine injection(高效液相色谱法测定左卡尼汀注射液的含量). Sci Technol Eng(科学技术与工程),2011,11(30):7494
7 SUN Yong-xu(孙永旭),LU Cong-xiao(陆丛笑),TANG Qi-ling(唐启令), et al.Simultaneous analysis of L-carnitine,acetyl-L-carnitine and propionyl-L-carnitine in human plasma by HPLC(高效液相色谱法同时测定人血浆中左卡尼汀、乙酰左卡尼汀和丙酰左卡尼汀的浓度). Chin Pharm J(中国药学杂志),2007,42(18):1425
8 Mardones C,Ríos A,Valcárcel M,et al.Separation of D/L-carnitine enantiomers by capillary electrophoresis.J Chromatogr A,1999,849(2):609
9 ZHAO Ya-ming(赵亚明),LI Ren(李任),WANG De-xin(王得新),et al .Establishment of the method for determination of plasma L-carnitine by high performance liquid chromatography(高效液相色谱法检测血浆左旋肉碱方法的建立).Chin J Clin Neurosci(中国临床神经科学),2006,14(5):528
10 Li K,Li W,Huang YF.Determination of free L-carnitinein human seminal plasma by high performance liquid chromatography with pre-column ultraviolet derivatization and its clinical application in male infertility.Clin Chim Acta,2007,378(5):159
11 XU Juan-juan(徐娟娟),LI Jie-mei(李杰梅),LIANG Shu-ming(梁淑明),et al .Analysis of the optical isomer of L-carnitine by chiral liquid chromatography(手性液相色谱法测定左旋肉碱中光学异构体的含量).Mod Food Sci Technol(现代食品科技),2010,26(3):311
12 ZHAO Rong(赵榕),SUN Kai-qi(孙开奇),LUO Ren-cai(罗仁才).An ion pair reversed phase high performance liquid chromatography method for determination of carnitine in health product(离子对反相高效液相色谱测定保健品中的肉碱).Chin J Food Hyg(中国食品卫生杂志),2002,14(4):25
13 Freimüller S,Altorfer H.A chiral HPLC method for the determination of low amounts of D-carnitine in L-carnitine after derivatization with(+)-FLEC.J Pharm Biomed Anal,2002,30(2):209