高三尖杉酯碱及其注射液杂质谱的研究

目的：建立LC-PDA-MS/MS法分析高三尖杉酯碱及其注射液的杂质谱。方法：采用Phenomenex C₁₈色谱柱，以0.01 mol·L^-1乙酸铵（0.1%乙酸）-乙腈为流动相，进行梯度洗脱，流速为0.3 mL·min^-1；通过光电二极管阵列检测器及电喷雾质谱检测法，采集杂质谱的PDA数据及各杂质的碎片离子数据，结合杂质的色谱行为及相关对照品的裂解规律，对未知杂质结构进行解析。结果：从原料药中分离出14个杂质，注射液中分离出15个杂质，并与对照品比对确定了其中1个杂质为三尖杉酯碱，另外分析了各杂质的归属与关联性。结论：建立的方法可有效分离高三尖杉酯碱及其注射液的杂质，通过对杂质谱的分析推断出各杂质的结构，为高三尖杉酯碱原料及其注射液质量控制提供一定的参考依据。

Objective: To establish an LC-PDA-MS/MS method for the determination of impurity profile in homoharringtonine and its injection. Methods: The HPLC separation was performed on a Phenomenex C₁₈ column by gradient elution,with a mobile phase consisting of 0.01 mol·L^-1 ammonium acetate (0.1% acetic acid)aqueous solution and acetonitrile at a flow rate of 0.3 mL·min^-1. The results were obtained by PDA and tandem mass spectrometry.The PDA data, debris ion data of all impurity in homoharringtonine were collected and the unknown impurity structure was elucidated according to chromatographic behavior of impurity and the fragmentation regularity of related reference substances. Results: The fourteen and fifteen impurity peaks were separated in homoharringtonine and its injection,respectively. One impurity peak could be identified as harringtonine by comparison with the behaviour of a reference substance in the same conditions. Also the attribution and association of each impurity were analyzed. Conclusion: The established method is effective for the separation of the related substances in homoharringtonine and its injection. The structure of the impurity can be determined by the analysis of impurity profile,which provide a reference for quality control of the homoharringtonine and its injection.

[1] 尹卫平,梁菊,吴文澜.抗癌天然药物研究进展[M].北京:科学出版社,2009:35 YIN WP,LIANG J,WU WL.Research Advances of the Natural Antitumor Drug[M].Beijing:Science Press,2009:35
[2] XU B.The influence of several anticancer agents on cell proliferation,differentiation and the cell cycle of murine erythroleukemia cells[J].Am J Chin Med,1981,9(4):268
[3] 胥彬.高三尖杉酯碱抗癌研究新进展[J].肿瘤药学,2013,3(3):162 XU B.Research advances of the anti-tumor effect of homoharringtonine[J].Antitumor Pharm,2013,3(3):162
[4] 中华人民共和国药典1990年版.二部[S].1990:588 ChP 1990.Vol Ⅱ[S].1990:588
[5] POWELL RG,WEISLEDER D,SMITH CR.Antitumor alkaloids from Cephalotaxus harringtonia:structure and activity[J].J Pharm Sci,1972,61(8):1227
[6] 马广恩,林隆泽,赵志远,等.三尖杉属植物中生物碱的研究Ⅰ.三尖杉中抗癌有效成分的分离和鉴定及新生物碱(+)-乙酰三尖杉碱[J].化学学报,1977,35(Z1):201 MA GE,LIN LZ,ZHAO ZY,et al.Studies on the alkaloids of Cephalotaxus Ⅰ.The isolation and characterization of antileukemic alkaloids together with a new alkaloid(+)-ace-tylcephalotaxine from Cephalotaxus fortunel[J].Acta Chim Sin,1977,35(Z1):201
[7] LIU YG.New Methods of Producing HHT:US,20100240887A1[P].2010-09-23
[8] LEE JY,KIM JH.Removal of residual methylene chloride from homoharringtonine by pre-treatment with ethanol[J].Process Biochem,2013,48(11):1809
[9] ZHOU MR,ZENG JP,WANG XM,et al.MiR-370 sensitizes chronic myeloid leukemia K562 cells to homoharringtonine by targeting Forkhead box M1[J].J Transl Med,2013,11:265
[10] CHEN P,YUAN Q,YANG H,et al.Homoharringtonine enhances bortezomib antimyeloma activity in myeloma cells adhesion to bone marrow stromal cells and in SCID mouse xenografts[J].Leuk Res,2017,57:119
[11] KANTARJIAN HM,O'BRIEN S,CORTES J.Homoharringtonine/omacetax-ine mepesuccinate:the long and winding road to food and drug administration approval[J].Clin Lymphoma Myeloma Leuk,2013,13(5):530
[12] 王倩,李玉峰.高三尖杉酯碱对伊马替尼耐药的慢性髓系白血病疗效的研究进展[J].中国实验血液学杂志,2017,25(2):596 WANG Q,LI YF.Research progress of homoharringtonine effect on IM-resistant chronic myelogenous leukemia[J].J Exp Hematol,2017,25(2):596
[13] 郭欢迎,王孝功,张晓敏,等.高三尖杉酯碱及其制剂有关物质的研究[J].中国药物警戒,2014,11(9):528 GUO HY,WANG XG,ZHANG XM,et al.Research for homobarringtonie and related substances of its preparations[J].Chin J Pharmacovigil,2014,11(9):528
[14] 胡昌勤.化学药品杂质控制的现状与展望[J].中国科学:化学,2010,40(6):679 HU CQ.Current situation and the trend in impurity control of chemical drugs[J].Sci Sin Chim,2010,40(6):679
[15] 沈卫阳,沙景阳,杨盈盈.HPLC-DAD/MS法测定辛伐他汀胶囊中有关物质[J].药物分析杂志,2014,34(6):1106 SHEN WY,SHA JY,YANG YY.Determination of related substances in simvastatin capsules by HPLC-DAD/MS[J].Chin J Pharm Anal,2014,34(6):1106
[16] 崔广青,周晓溪,秦爱芳,等.LC-MS/MS法分析注射用哌拉西林钠的杂质谱[J].药物分析杂志,2015,35(9):1612 CUI GQ,ZHOU XX,QIN AF,et al.Analysis of impurity spectra in piperacillin sodium for injection by LC-MS/MS[J].Chin J Pharm Anal,2015,35(9):1612
[17] HE J,CHEUNG AP,WANG E,et al.Stability-indicating LC assay of and impurity identification in homoharringtonine samples[J].J Pharm Biomed Anal,2000,22(3):541
[18] CHOI YH,YOO KP,KIM J.HPLC-electrospray ionization-MS-MS analysis of Cephalotaxus harringtonia leaves and enhancement of the extraction efficiency of alkaloids therein by SFE[J].J Chromatogr Sci,2003,41(2):67