建立HILIC-UPLC联用分析法测定血浆中葫芦巴碱及在药动学研究中的应用(英文)

Development of a hydrophilic interaction chromatography-UPLC assay to determine trigonelline in rat plasma and its application in a pharmacokinetic study

目的:葫芦巴碱是咖啡豆中含量较多的生物碱之一,为了研究其药代动力学特征,本研究建立了一种简便、灵敏、准确的亲水相互作用色谱-超高效液相联用分析方法,并研究了葫芦巴碱和咖啡豆甲醇提取物的药代动力学。方法:大鼠灌胃和尾静脉注射给予葫芦巴碱和咖啡豆甲醇提取物后,眼眶静脉丛取血,血液样本采用亲水相互作用色谱-超高效液相联用分析法进行分析,药代动力学参数按非房室模型,通过药动学软件87/97进行计算。结果:对所建立的方法进行了方法学的考察,线性范围为0.12–100μg·mL-1,最低定量下限为0.12μg·mL-1,日内日间精密度、准确度、提取回收率和稳定性等均达到体内药物分析要求。葫芦巴碱口服和尾静脉注射给药后,AUC(0-∞)分别为(4066.83±1244.41)和(3544.29±908.80)min·μg·mL-1,生物利用度达到57.37%。咖啡豆甲醇提取物口服后,AUC(0-∞)为(4566.75±1435.64)min·μg·mL-1,绝对生物利用度为64.42%,相对生物利用度为112.29%。结论:所建立的亲水作用色谱-超高效液相联用分析法简单、准确、重现性好;葫芦巴碱无论是纯品还是在咖啡豆甲醇提取物中都有较好的生物利用度,而且没有明显的差异。

AIM： Trigonelline （Tr） is the second most abundant alkaloid in coffee beans. This study developed an assay combining hydrophilic interaction chromatography with ultra performance liquid chromatography （HILIC-UPLC） for the quantification of Tr in rat plasma to determine its pharmacokinetic behavior. METHODS： After the administration of Tr by gavage as well as intravenous injection and that of methanol extract of coffee beans （MECB） orally, blood samples from the experimental rats were analyzed using the HILIC-UPLC assay. Pharmacokinetic parameters were determined using the standard non-compartmental method and calculated using Practical Pharmacokinetic Program Version 87/97. RESULTS： The HILIC-UPLC assay was validated with the linear range of 0.12-100 μg·mL^-1 and a lower limit of quantitation of 0.12 μg·mL^-1 Its accuracy, precision, recovery, and stability were within acceptable limits. The AUC（0-ω） （where AUC is the area under the plasma concentration-time curve） values were determined to be （4 066.83 ± 1 244.41） and （3 544.29 ± 908.80） min·μg·mL^-1 after Tr was orally and intravenously administered, respectively. It was （4 566.75 ± 1 435.64） min·μg·mL^-1 after MECB was orally administered. The absolute bioavailability of Tr alone reached 57.37%, whereas that of Tr in MECB was 64.42%. The relative bioavailability of the alkaloid was 112.29%. CONCLUSIONS： The HILIC-UPLC assay for Tr determination is simple and accurate, and also exhibits good reproducibility. The bioavailability of stand-alone Tr and that of Tr in MECB were both good. Tr alone and that in MECB orally administered did not exhibit any significant difference.