HPLC法测定大鼠肺组织中双(对-氟苄基)三硫醚及其降解产物的浓度

Determination of bis(p-fluorobenzyl)trisulfide and bis(p-fluoro-benzyl) disulfide in lungs of rat by HPLC

目的：建立一种测定大鼠肺组织中双（对-氟苄基）三硫醚（BFTS）及其降解产物双（对-氟苄基）二硫醚（BFDS）的RP—HPLC分析方法。方法：在0．2g大鼠肺组织中加入正己烷-异丙醇（95：5，v／v）5．0ml，11．50μg／ml内标（苄基二硫醚）溶液20μl，制成匀浆，离心后取上清液4．0ml，真空抽干，残渣加流动相（乙腈：水=65：35，v／v）150μl复溶，离心后取上清液进行HPLC分析。HPLC分析条件为：SB—C18柱，流动相为乙腈-水（65：35，v／v）；流速：1．0ml／min；柱温：30℃；检测波长：220nm；进样量：100μl。结果：BFTS和BFDS在0．04712～14．78μg／g（r=0．999）及0．04831～23．96μg／g（r=0．999）范围内呈线性关系，定量限分别为0．04712μg／g和0．04831μg／g，回收率分别为95．71％～107．2％和90．00％～110．5％，精密度RSD均小于10％。该方法已成功地用于大鼠静脉注射12．5mg／kg BFTS后，肺组织中BFTS及BFDS的含量测定。结论：该法准确、灵敏、可靠，可用于BFTS及BFDS的组织分布研究。

Objective： To establish an HPLC trisulfide （BFTS） and bis （p-fluorobenzyl）disulfide extract solvent （n-he disulfide （internal sta centrifugation, 4.0 ml reconstituted in mobil column using acetonit method for analysis of bis （p-fluorobenzyl） xane ： isopropyl alcohol = 95 ： ndard） were added to 0.2 g lu （BFDS） in the lungs of rat. M 5,v/v） and 20 μl of 11. 50 ng sample followed by homog of supernatant was separated and vapo e phase rile and for HPLC analysis. The HPLC rized analy water （65 ：35,v/v） as mobile phase etbods： 5.0 ml 〉g/ml dibenzyl enization. After to dryness, and the residue was sis was performed on an SB C18 with a flow rate of 1.0 ml/min with UV detection at 220 nm. Results ： The calibration curves for BFTS and BFDS in sample were linear over the concentration ranges of 0. 04712-14.78 μg/g（r=0. 999） and 0. 04831-23.96 μg/g （r= 0. 999）,respectively. The limits of quantification were 0. 04712 μg/g and 0. 04831 μg/g for BFTS and BFDS,respectively. The assay recoveries for BFTS and BFDS ranged from 95.71%- 107.2% and 90. 00%-110.5% ,respectively. The precisions were obtained with RSD of 〈10%. The developed method was successfully applied to study the content of BFTS and BFDS in the lungs of rats after intravenous injection of 12.5 mg/kg BFTS. Conclusion： The method developed is simple ,selective ,repeatable and accurate, which can be applied to study the tissue distribution of BFTS and BFDS.