液相微萃取-非水后萃取-高效液相色谱法测定大鼠体内厚朴酚与和厚朴酚的浓度

Determination of magnolol and honokiol in rats by liquid-phase microextraction with nonaqueous back extraction coupled with HPLC

目的:建立液相微萃取-非水后萃取(LPME/NBE)-高效液相色谱法同时测定大鼠体内厚朴酚及和厚朴酚浓度的方法。方法:利用自制的液相微萃取装置,以正丁醇为萃取溶剂,以600 r·min^(-1)转速萃取10min,对大鼠血浆、肝脏和肾脏生物样品中的厚朴酚与和厚朴酚进行分离、萃取、纯化。色谱条件:ODS为色谱柱.甲醇-水(82:18)为流动相,294 nm为检测波长。结果:厚朴酚与和厚朴酚线性范围分别为0.15～30.0mg·L^(-1)和0.10～30.0 mg·L^(-1),r>0.900;RSD<7.5%;平均回收率分别为93.7%～114%和90.5%～109%,血浆中检出限分别为30μg·L^(-1)和20μg·L^(-1)。肝脏中分别为25μg·L(-1)和15μg·L(-1),肾脏中均为10μg·L(-1)。结论:首次提出液相微萃取-非水后萃取方法.并将其成功应用于中药厚朴酚与和厚朴酚在大鼠体内的浓度测定。液相微萃取-非水后萃取能有效去除生物样品中干扰厚朴酚与和厚朴酚测定的内源性杂质.提高选择性。

OBJECTIVE To develop a method for determining the concentration of magnolol and honokiol in biological matrices by liquid-phase micro-extraction with nonaqueous back extraction （LPME/NBE） combined with high performance liquid chromatography （HPLC）. METHODS By self-made liqui＆phase micro-extraction apparatus self-made, n-butanol as extraction solvent, magnolol and honokiol were extracted 10 min from biological samples at 600 r,min^-1. Alter extracted, the acceptor solution was injected into HPLC for detecting. The chromatographic column was ODS, the mobile phase consisted of methanol-water （82： 18）. The UV detection wavelength was 294 nm RESULTS The linear ranges of magnolol and honokiol were 0. 15 - 30. 0 mg·L^-1 and 0.10 - 30.0 mg·L^-1 respectively, coefficients of correlation （r） were better than 0.90, relative standard deviation （RSD） was lower than 7. 50/00. The average relative recoveries were 93.7% - 114% and 90. 5% - 109% respec- tively. The limit of detection of magnolol in plasma, liver and kidney samples were 30, 25, 10 μg. L^-1 respectively, meanwhile, the limit of honokiol in these biological samples were 20, 15, 10 μg·L^-1. CONCLUSION Liquid-phase microextraction with nonaqueous back extraction was adopted for the first time. This method was applied successfully to simultaneous determination of magnolol and honokiol in biological samples. With this method, some endogenous substances in matrices interfering the determination of magnolol and honokiol were eliminated efficiently, and the higher selectivity was obtained.