色谱

色谱 ›› 2012, Vol. 30 ›› Issue (11): 1183-1187.DOI: 10.3724/SP.J.1123.2012.06039

• 研究论文 • 上一篇    下一篇

高效液相色谱法同时测定离体角膜灌流液中8个普萘洛尔衍生物

伍海涛1*, 陈传兵2, 王宁生1, 宓穗卿1, 廖南英1   

  1. 1. 广州中医药大学临床药理研究所, 广东 广州 510405; 2. 广州中医药大学中药学院, 广东 广州 510405
  • 收稿日期:2012-06-28 修回日期:2012-08-02 出版日期:2012-11-28 发布日期:2012-11-21
  • 通讯作者: 伍海涛,博士,研究方向为临床药理和眼科药理. Tel: (020)36585405, E-mail: znwht@sina.com.cn.
  • 基金资助:

    广东省“211工程”三期重点学科建设项目(粤发改社［2009］972号)、广东省建设中医药强省课题(2010373)、广东省自然科学基金项目(S2011040003546)和广东省科技计划项目(2011B031700069).

Simultaneous determination of eight derivatives of propranolol in cornea perfusate in vitro by high performance liquid chromatography

WU Haitao1*, CHEN Chuanbing2, WANG Ningsheng1, MI Suiqing1, LIAO Nanying1   

  1. 1. Institute of Clinical Pharmacology, Guangzhou University of Chinese Medicine, Guangzhou 510405, China; 2. School of Chinese Pharmaceutical Science, Guangzhou University of Chinese Medicine, Guangzhou 510405, China
  • Received:2012-06-28 Revised:2012-08-02 Online:2012-11-28 Published:2012-11-21

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346

被引次数

2

摘要： 为了解促渗透剂对不同物质的促渗透能力,建立了采用高效液相色谱分离测定8个普萘洛尔衍生物的分析方法。采用盒式给药技术在离体角膜上皮侧给予普萘洛尔衍生物,内皮侧接收灌流液,用甲醇沉淀蛋白、高速离心后进样,采用Agilent Zorbax Extend色谱柱,以乙腈(A)-0.03%(v/v)磷酸水溶液(B)为流动相,在3%A～20%A间梯度洗脱,柱温30 ℃,检测波长205 nm。结果表明,该方法可在31 min内实现离体角膜灌流液中8个衍生物的完全分离和定量,线性范围为0.2(0.1)～40.0 μmol/L,线性关系良好,相关系数均不小于0.9970。其加样回收率为91.12%～105.73%,日内精密度(相对标准偏差(RSD))为1.00%～11.63%,日间精密度为1.18%～18.58%,样本冻存、冻融稳定性良好。该法适用于离体角膜灌流、细胞小室培养等跨膜吸收研究中普萘洛尔衍生物的检测。

关键词: 高效液相色谱, 角膜, 普萘洛尔, 衍生物

Abstract: A high performance liquid chromatographic method was developed for the simultaneous determination of eight derivatives of propranolol. Cassette dosing method was used in the epithelium side of cornea in vitro to get the effect of penetrant, and the perfusate was collected in the side of endothelium. The protein in the sample was precipitated and discarded by high speed centrifugation before injection. An Agilent Zorbax Extend column (150 mm×3 mm, 5 μm) was used at 30 ℃. The mobile phase system contained acetonitrile and 0.03% (v/v) phosphoric acid aqueous solution and the percentage of acetonitrile changed between 3% and 20% (v/v) in a linear gradient elution. The samples were detected by an ultraviolet (UV) detector at 205 nm. The results showed that the eight derivatives of propranolol were completely separated and determined in 31 min. The correlation coefficients were above 0.9970 and good linear relationships were obtained in the range of 0.2(0.1)~40.0 μmol/L. Under the optimized conditions, the recoveries of the derivatives were in the range of 91.12%~105.73%. The intra-day relative standard deviations (RSDs) were in the range of 1.00%~11.63%, and the inter-day RSDs were in the range of 1.18%~18.58%. The sample showed stability under room temperature, freeze and three cycles of freeze-thaw conditions. This method is fast and accurate for the quantitative analysis of the derivatives of propranolol in transmembrane absorption such as cornea perfusion in vitro or transwell cell system.

Key words: cornea, derivatives, propranolol, high performance liquid chromatography (HPLC)