Hanada K, Nagai N, Ogata H
Department of Biopharmaceutics, Meiji College of Pharmacy, Tokyo, Japan.
J Chromatogr B Biomed Appl. 1995 Jan 6;663(1):181-6. doi: 10.1016/0378-4347(94)00415-2.
A quantitative analytical method for measuring unchanged cisplatin (CDDP) and high- and low-molecular-mass metabolites (fixed and mobile metabolites) in rat kidney and liver was developed. Unchanged CDDP, separated from fixed and mobile metabolites in tissue homogenates by consecutive procedures of fractionation and ultrafiltration, was determined by high-performance liquid chromatography (HPLC) with post-column derivatization. Although unchanged CDDP was found to be partly metabolized to fixed metabolites during the preparation of cytosolic ultrafiltrates, the recovery of unchanged CDDP gave a constant value (about 70%), which was independent of tissue type and CDDP concentration (from 1 to 10 micrograms/ml). The detection limit for unchanged CDDP in the cytosolic ultrafiltrate was 20 ng/ml, corresponding to a concentration detection limit of 65 ng Pt per g of tissue in the kidney and liver. The concentrations of fixed and mobile metabolites were determined as platinum concentrations in the tissue homogenate and in the cytosolic ultrafiltrate using atomic absorption spectrometry after correcting for transformation of unchanged CDDP to fixed metabolites. The distribution of unchanged CDDP, mobile metabolites and fixed metabolites in rat kidney and liver, after bolus injection of CDDP (5 mg/kg), was determined using this method.
建立了一种定量分析方法，用于测定大鼠肾脏和肝脏中未变化的顺铂（CDDP）以及高分子量和低分子量代谢物（固定和流动代谢物）。通过分级分离和超滤的连续步骤，将组织匀浆中未变化的CDDP与固定和流动代谢物分离，然后采用柱后衍生的高效液相色谱（HPLC）法进行测定。尽管发现在制备胞质超滤液过程中，未变化的CDDP有部分代谢为固定代谢物，但未变化的CDDP回收率为恒定值（约70%），且与组织类型和CDDP浓度（1至10微克/毫升）无关。胞质超滤液中未变化的CDDP检测限为20纳克/毫升，相当于肾脏和肝脏中每克组织65纳克铂的浓度检测限。在对未变化的CDDP转化为固定代谢物进行校正后，使用原子吸收光谱法测定组织匀浆和胞质超滤液中固定和流动代谢物的浓度，以铂浓度表示。采用该方法测定了大鼠静脉注射CDDP（5毫克/千克）后，未变化的CDDP、流动代谢物和固定代谢物在肾脏和肝脏中的分布情况。
