HPLC-UV法测定大鼠体内S-腺苷蛋氨酸浓度及其药动学研究

目的：建立HPLC-UV法测定大鼠血浆中S-腺苷蛋氨酸（S-Adenosylmethionine,SAMe）浓度,并用此法研究静脉给药后的大鼠体内药代动力学特征。方法：色谱柱：大连依利特KromasilC18柱（4.6mm×200mm,5μm）,流动相：甲醇∶0.3%三氟乙酸溶液（含1%磷酸二氢钠盐）=2∶98,等浓度洗脱,紫外检测波长：260nm,流速：0.6mL/min,柱温：30℃。大鼠（n=6）经静脉给予140mg/kg SAMe并测定血浆药物浓度,采用DAS 2.0软件进行药动分析。结果：SAMe能完全分离,线性范围：10～2000μg/mL,回归方程为Y=30241 X＋256204,r=0.9991,日内精密度分别为2.6%,3.6%,3.9%;日间精密度分别为6.3%,4.9%,7.9%;回收率分别为107%,101%,98.3%。静脉给予140mg/kg SAMe药动学参数,血浆药物浓度-时间曲线下面积AUC（0-t）为（569.52±174.87）mg·L-1·h,半衰期（t1/2）为（2.03±0.96）h,清除率（CL）为（0.21±0.13）L·h-1·kg-1,表观分布容积（Vd）为（0.53±0.31）L/kg。结论：本实验方法专属性好,灵敏度高,可靠性强,可用于SAMe药动学分析。

Determination and pharmacokinetics of S-Adenosylmethionine in rats plasma by HPLC-UV

AIM： To establish an HPLC-UV assay for determining plasma concentration of S-Adenosylmethionine（SAMe） and to study its pharmacokinetics in rats.METHODS： The chromatographic conditions were as follows： a Kromasil C18 column（4.6 mm×200 mm,5 μm）,a flow rate of 0.6 mL/min for the mobile phase of methanol/0.3% trifluoroacetic acid（V/V）（dissolve 1% phosphate buffer）（2∶98）,an isocratic elution mode was carried out at column temperature（30 ℃） and the UV detection at the wavelength of 260 nm.140 mg/kg SAMe was administrated by intravenous and plasma concentration were determined by HPLC-UV assay.RESULTS：SAMe was successfully separated.The calibration curve showed a good linear regression within the range of 10-2000 μg/mL.The regression equation was Y=30241X＋256204,r=0.9991.The precision of intra-assay and inter-assay were 2.6%,3.6%,3.9% and 6.3%,4.9%,7.9%,respectively.The average extraction recovery was 107%,101%,98.3%,respectively.The pharmacokinetic parameter were AUC（0-t）（569.52±174.87） mg·L-1·h,t1/2（2.03±0.96） h,CL（0.21±0.13） L·h-1·kg-1,Vd（0.53±0.31） L/kg,respectively.CONCLUSION： This method was proved to be simple and efficient with excellent specificity,sensitivity,precision,recovery and can be applied to pharmacokinetic investigation of SAMe.