摘要
目的建立HPLC测定人血浆中总的和游离麦考酚酸(MPA)及其代谢物葡糖苷酸结合物(MPAG)含量的方法。方法游离和总的药物含量测定的预处理方法分别采用超滤法和乙腈蛋白沉淀法。色谱条件:KromasilC8(4.6mm×150mm,5μm);柱温:30℃。流动相:乙腈-0.1%三氟醋酸(55:45,pH2.2),流速:1.2mL·min-1。柱后添加0.2mol·L-1NaOH溶液,流速0.15mL·min-1。MPAG采用紫外检测,检测波长为295nm;MPA采用荧光检测,荧光激发波长342nm,发射波长425nm。结果血浆内源性杂质和常用合并用药对样品测定无干扰,MPA,MPAG的总质量浓度分别在0.1-40和10-250mg·L-1,游离质量浓度分别在0.010-1.0和2.5-100mg·L-1内线性关系良好。MPA,MPAG的日内、日间RSD均小于10%,样品提取后在24h内及3次冻融后稳定性良好。结论本方法简便快速、结果准确可靠,可用于MPA和MPAG的药动学研究及常规的血药浓度监测。
OBJECTIVE To establish a HPLC method for determining total and free concentration of mycophenolic acid （MPA） and its glucuronide （MPAG） in human plasma. METHODS Plasma samples were treated by ultra-filtration and protein precipitation for the determination of free and total analytes, respectively. Kromasil C8 （4.6 mm ×150 mm, 5μn） was used as the analytical column and maintained at 30 ℃. The mobile phase consisted of a mixture of methanol：0.1% trifluoroacetic acid （55：45,pH 2.2） at a flow rate of 1.2 mL·min^-1. Regarding to the postcolumn derivatization, the other HPLC pump was added to deliver 0.2 mol· L^- 1 NaOH at a flow rate of 0.2 mL· min^-1, MPAG was determined by UV absorbance and the detection wavelength was set at 295 nm while MPA was measured by fluorescence detection with λex 342 nm and λem 425 nm.RESULTS The endogenous interference and commonly co-medicated medicine did not interfere with the analytes determination. The alibration curves of total MPA and MPAG were in the ranges of 0.1 - 40 and 10 - 250 mg· L^- 1, as well as free MPA and MPAG of 0.01 - 1.0 and 2.5 - 100 mg·L^- 1 ,respectively. The RSDs of within-nm and between-nm were less than 10%. The samples were stable for 24 h after being processed at room temperature and the three-cycle of freezing and thawing test. CONCLUSION The current established method can be used to determine the concentrations of total and free MPA and MPAG in human plasma and it can also be applied to the clinical pharmacy research and routine therapeutic drug monitoring.
出处
《中国药学杂志》
EI
CAS
CSCD
北大核心
2005年第17期1339-1343,共5页
Chinese Pharmaceutical Journal
基金
上海市自然科学基金(03ZR14010)
