反相高效液相色谱法测定家犬体内甲氧氯普胺的血药浓度

RP-HPLC Determination of Metoclopramide in Dog Plasma

目的:建立测定甲氧氯普胺血浆浓度的高效液相色谱法。方法:血浆样品在碱性条件下用乙醚提取,盐酸曲马多为内标,采用 Hypersil-C_(18)(4.6mm×200mm,5μm)柱,流动相为甲醇-乙腈-醋酸盐缓冲液(pH 4.0)-三乙胺(7:18:75:0.1),检测波长273nm,甲氧氯普胺与内标的保留时间分别为4.9min 和7.1min。结果:线性范围为5-100ng·mL^(-1)(r=0.9993);最低检测浓度为1ng·mL^(-1);日内、日间精密度 RSD 分别为2.4%-6.8%,3.6%-8.6%(n=5);低、中、高3个浓度的提取回收率分别为81.5%,84.6%,80.4%;方法回收率为96.6%,98.5%,101.2%。结论:本方法灵敏度高,简便易行,能满足甲氧氯普胺的药物动力学研究。

Objective:To establish an RP-HPLC method for determination of metoclopramide (MCL) concentration in dog plasma.Methods:Metoclopramide was extracted with ether under the alkaline condition.Hypersil-C_(18) (4.6 mm×200 mm,5 μm)column was employed and the mobile phase consisted of a mixture of methanol-acetonitrile -acetate buffer(pH 4.0)-triethylamine (7:18:75:0.1) with a flow rate of 1.0 mL·min^(-1).Tramadol hydrochlo- ride was used as an internal standard.Results:A linear calibration curve of MCL in plasma was obtained in the con- centration from 5.0 ng·mL^(-1)to 100.0 ng·mL^(-1)(r=0.9993).The minimal detectable drug plasma concentration was 1 ng·mL^(-1).Intra-day and inter-day RSD were 2.4%-6.8% and 3.6%-8.6%,respectively.The ex- tracted recoveries were 81.5%,84.6%,80.4%,and method recovery were 96.6%,98.5%,101.2% at low,mid- dle and high concentrations,respectively.Conclusion:A simple,sensitive and selective HPLC method for determi- nation of metoclopramide concentration in plasma was established.