Woolf E J, Au T, Kasper M, Constanzer M, Matuszewski B
Merck Research Laboratories, Department of Drug Metabolism, West Point, PA 19486.
J Chromatogr A. 1994 Feb 4;660(1-2):307-12. doi: 10.1016/0021-9673(94)85126-3.
A method for the simultaneous determination of a topical carbonic anhydrase inhibitor, L-693,612, and two of its potential metabolites in human whole blood is described. The analytes are isolated from the matrix via liquid-liquid extraction with a mixture of toluene, ethyl acetate and isopropanol (49:50:1, v/v/v). The analytes are then back extracted into dilute phosphoric acid prior to injection into the HPLC system. A cyano column (Zorbax SB-CN, 150 x 4.6 mm) with a mobile phase of phosphoric acid(0.085%)-acetonitrile (73.5:26.5) containing 10 mM sodium decane sulfonate and adjusted to pH 3 is used for the analysis. Detection is based on UV absorbance at 252 nm. The assay was found to be linear in the concentration range of 5-500 ng/ml for each analyte when 1-ml aliquots of whole blood were extracted.
本文描述了一种同时测定人全血中局部碳酸酐酶抑制剂L-693,612及其两种潜在代谢物的方法。通过用甲苯、乙酸乙酯和异丙醇（49:50:1，v/v/v）的混合物进行液-液萃取，从基质中分离出分析物。然后在注入HPLC系统之前，将分析物反萃取到稀磷酸中。使用氰基柱（Zorbax SB-CN，150×4.6 mm），流动相为含10 mM癸烷磺酸钠且pH值调至3的磷酸（0.085%）-乙腈（73.5:26.5），基于252 nm处的紫外吸光度进行检测。当萃取1 ml全血等分试样时，发现该测定法对每种分析物在5 - 500 ng/ml的浓度范围内呈线性。
