Wu Chunyu, Njar Vincent, Brodie Angela, Borenstein Michael, Nnane Ivo
Department of Pharmaceutical Sciences, School of Pharmacy, Temple University, Philadelphia, PA 19140, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Oct 25;810(2):203-8. doi: 10.1016/j.jchromb.2004.07.028.
A simple reversed phase high performance liquid chromatographic (HPLC) method was developed for the separation and quantification of a novel retinoic acid metabolism inhibitor, 4-(1H-imidazol-1-yl)retinoic acid (VN/14-1RA), and other retinoids in rat plasma. VN/14-1RA, alone or in combination with ATRA, is effective at inhibiting the proliferation of prostate and breast cancer cell lines in vitro. Aliquots of rat plasma were spiked with the retinoids followed by addition of acetonitrile for precipitation of plasma proteins. The decanted supernatant was evaporated under a stream of nitrogen and reconstituted in acetonitrile. Analysis was accomplished by injection of an aliquot of the reconstituted sample into an HPLC system consisting of a Zorbax Rx-C18 column and a diode array detector. A mobile phase composed of ammonium acetate (0.1 M), acetic acid solution (2% (v/v)) and methanol at a flow rate of 1.0 mL/min was used for gradient elution. The recoveries for all compounds ranged from 65 to 85% regardless of the concentrations examined. The HPLC assay was linear over the range 0.10-5.0 microg/mL (CV < 10%) with a limit of quantification of 100 ng/mL for VN/14-1RA. A one-compartment model with apparent first-order elimination was used to describe the plasma concentration-time profile for VN/14-1RA after intravenous administration. The mean terminal elimination half-life (t(1/2)) was 19.0 +/- 3.2 min. This HPLC method is useful for the analysis and evaluation of the pharmacokinetics of VN/14-1RA in rats.
建立了一种简单的反相高效液相色谱（HPLC）方法，用于分离和定量大鼠血浆中的新型维甲酸代谢抑制剂4-（1H-咪唑-1-基）维甲酸（VN/14-1RA）及其他类视黄醇。VN/14-1RA单独或与全反式维甲酸（ATRA）联合使用时，在体外可有效抑制前列腺和乳腺癌细胞系的增殖。取等分的大鼠血浆，加入类视黄醇，随后加入乙腈沉淀血浆蛋白。将倾析得到的上清液在氮气流下蒸发，并用乙腈复溶。通过将复溶样品的等分试样注入由Zorbax Rx-C18柱和二极管阵列检测器组成的HPLC系统来完成分析。使用由醋酸铵（0.1 M）、醋酸溶液（2%（v/v））和甲醇组成的流动相，流速为1.0 mL/min进行梯度洗脱。无论所检测的浓度如何，所有化合物的回收率在65%至85%之间。HPLC测定在0.10 - 5.0 μg/mL范围内呈线性（CV < 10%），VN/14-1RA的定量限为100 ng/mL。采用具有表观一级消除的单室模型来描述静脉给药后VN/14-1RA的血浆浓度-时间曲线。平均末端消除半衰期（t(1/2)）为19.0 +/- 3.2分钟。该HPLC方法可用于分析和评估VN/14-1RA在大鼠体内的药代动力学。
