中空纤维离心超滤法检测人血浆中氨茶碱的游离药物浓度

Determination of unbound theophylline in human plasma by hollow fiber centrifugal ultrafiltration

目的：建立一个准确、简单、快速的分析人血浆中游离氨茶碱浓度的方法,并应用于临床治疗药物监测。方法：0.5 ml血浆样品经过中空纤维离心超滤（HFCF-UF）处理,以甲醇-10 mmol·L-1乙酸铵缓冲溶液（15/85,V/V）为流动相,采用Diamonsil C18色谱柱分离,流速为1.0ml·min-1,检测波长为275 nm,进样量为20μl。结果：血浆中游离氨茶碱浓度在0.25-25μg·ml-1范围内线性关系良好,最低定量限0.1μg·ml-1,日内、日间RSD分别小于1.1%和1.3%。结论：该方法简单、准确、重现性好,特别适合基层开展临床游离药物浓度的监测。该方法成功的应用于临床游离氨茶碱浓度的监测,为临床游离氨茶碱治疗药物监测提供了一个可靠的分析平台和进一步阐述游离药物浓度与药物疗效或毒性之间的关系奠定了基础。

OBJECTIVE To develop an accurate,simple and rapid method for determination of unbound theophylline in human plasma,and apply it for monitoring clinical therapeutic drugs.METHODS A volume of 0.5 ml plasma was subjected to hollow fiber centrifugal ultrafiltration（HFCF-UF）,and ultrafiltrates were then separated on a Diamonsil C18 column.Mobile phase consisted of methanol/10 mmol·L-1 ammonium acetate buffer solution（15/85,V/V）,at a flow rate of 1.0ml·min-1.Detention wavelength was set at 275 nm and injection volume was 20μl.RESULTS Unbound theophylline showed a good linear range in0.25-25μg·ml-1.The lowest detection limit was 0.1μg·ml-1.Intra-day and inter-day RSD values were less than1.1% and 1.3%,respectively.CONCLUSION The method is simple,accurate and reproducible,especially for conduction of clinical therapeutic drug monitoring at fundamental hospitals.Validated method has been successfully applied for patient samples for routine therapeutic drug monitoring of theophylline,and it therefore may become a feasible platform for analysis of unbound drug and provide theoretical basis for elucidation of relationship between unbound concentration and efficacy or toxicity in future study.