摘要
目的建立新药甲磺酸帕珠沙星(PZLX)对映体手性流动相HPLC拆分方法。方法通过探讨流动相pH值、有机改性剂、手性配合剂、反相固定相对手性拆分的影响,确定甲醇-苯丙氨酸溶液(L-苯丙氨酸1.32g与硫酸铜1g,加水1000ml,氢氧化钠试液调节pH值为3.5)(30∶70)为手性流动相,在ODS柱上拆分PZLX对映体,332nm波长检测。结果两对映体良好分离,分离度1.9以上,S、R异构体回归方程分别为:Y=1.38×105X-130.9(r=0.9998)和Y=1.26×105X-60.6(r=0.9998),最低检出限分别为1.2和1.5ng,RSD为0.2%以下。结论所建立的方法简便易行,克服了该类方法灵敏度及重现性不如手性固定相法(CSP)的不足,可用于新药PZLX光学纯度的质量控制及其生物体内代谢立体选择性的研究。
Objective To establish an HPLC method for separating and determining the enantiomers of pazufloxacin mesylate.Methods By optimizing factors which affect the chiral separation,the pH value of the mobile phase, the organic modifiers,chiral additives,and stationary phases,the optimum condition for chiral separation were selected.The enantiomers separation was carried out on Shimandzu ODS column(150mm×6.3mm,10μm),and methanol-phenylalanine solution(containing 8mmol/L L-phenylalanine and 6.3 mmol/L copper sulphate,adjusted to pH=3.5 with sodium hydroxide solution)(30∶70) as the chiral mobile phase,detecting on wavelength at 332nm. Results The two enantiomers could be baseline separated,and the resolution was higher than 1.9.Calibration graphs were constructed in the range of 25～250μg/ml for S-(-)-and R-(+)-enantiomer,respectively.The linear regression equations are:Y=1.38×105X-130.9 (r=0.9998)and Y=1.26×105X-60.6 (r=0.9998),respectively.The detection limits obtained by S/N=3 were 1.2ng for S-(-)-and 1.5ng for R-(+)-enantiomer,respectively.Conclusions The proposed method was simple,sensitive,accurate and reproducible,and it can be used for the quality control of the pazufloxacin mesylate enantiomers.
出处
《中国抗生素杂志》
CAS
CSCD
北大核心
2004年第1期19-22,共4页
Chinese Journal of Antibiotics
基金
国家自然科学基金资助项目(20375010)
河北省基金资助项目(202096
0224501-D)
科技部资助(2002CCA3100
KJCX2-H4)
教育部优秀青年教师资助
