Nielsen Flemming, Christensen Mette M H, Brøsen Kim
Clinical Pharmacology, Institute of Public Health, University of Southern Denmark, Odense, Denmark.
Ther Drug Monit. 2014 Apr;36(2):211-7. doi: 10.1097/FTD.0b013e3182a4598a.
We describe an analytical method for the quantification of the widely used antihyperglycemic agent, metformin, in human plasma and urine. The separation was performed using isocratic hydrophilic interaction liquid chromatography on a Luna hydrophilic interaction liquid chromatography column (125 × 4.6 mm, 3 μm). The sample preparation was accomplished by solid-phase extraction. Validation of the method was performed in the range 10-2000 ng/mL for plasma and 5-30 mcg/mL for urine. The methods were linear within the investigated range (r(2) > 0.988). Within-day repeatability ranged from 3.1% to 7.5% in plasma and 1.6% to 6.2% in urine. Between-day reproducibility ranged from 2.9% to 5.3% in plasma and 0.6% to 1.8% in urine. The inaccuracy expressed as bias ranged from -3.1% to 1.9% in plasma and from -7.2% to 0.7% in urine. The lower limit of quantification for metformin in plasma was 5 ng/mL and in urine was 40 ng/mL. The method was therefore considered to be precise, accurate, reproducible, and sensitive enough to be appropriate for pharmacokinetic studies of metformin. The applicability of the method for human pharmacokinetic studies was demonstrated by dosing a healthy male volunteer with 500-mg metformin hydrochloride as a single oral dose; plasma and urine concentrations were measured for 24 hours.
我们描述了一种用于定量测定人血浆和尿液中广泛使用的抗高血糖药物二甲双胍的分析方法。分离采用等度亲水相互作用液相色谱法，使用月旭亲水相互作用液相色谱柱（125×4.6 mm，3μm）。样品制备通过固相萃取完成。该方法在血浆10 - 2000 ng/mL和尿液5 - 30 mcg/mL范围内进行验证。在所研究的范围内方法呈线性（r(2)>0.988）。日内重复性在血浆中为3.1%至7.5%，在尿液中为1.6%至6.2%。日间重现性在血浆中为2.9%至5.3%，在尿液中为0.6%至1.8%。以偏差表示的不准确度在血浆中为 - 3.1%至1.9%，在尿液中为 - 7.2%至0.7%。二甲双胍在血浆中的定量下限为5 ng/mL，在尿液中为40 ng/mL。因此，该方法被认为精确、准确、可重现且足够灵敏，适用于二甲双胍的药代动力学研究。通过给一名健康男性志愿者单次口服500 mg盐酸二甲双胍来证明该方法在人体药代动力学研究中的适用性；测定24小时内的血浆和尿液浓度。
