Weng N, Lee J W, Hulse J D
Harris Laboratories, Inc., Lincoln, NE 68501.
J Chromatogr B Biomed Appl. 1994 Apr 1;654(2):287-92. doi: 10.1016/0378-4347(94)00010-7.
An isocratic HPLC method was developed and validated for the quantitation of methocarbamol in human plasma. Methocarbamol and internal standard in 200 microliters of human plasma were extracted with ethyl acetate, evaporated to dryness and reconstituted in water. Separation was achieved on a reversed-phase C18 column with a mobile phase of methanol-0.1 M potassium phosphate monobasic-water (35:10:55, v/v/v). The detection was by ultraviolet at 272 nm. Linearity was established at 1-100 micrograms/ml (r > 0.999). The limit of quantitation was designed as 1 microgram/ml to suit pharmacokinetic studies. Inter-day precision and accuracy of the calibration standards were 1.0 to 3.6% coefficients of variance (C.V.) and -2.0 to +1.6% relative error (R.E.). Quality controls of 3, 20 and 70 micrograms/ml showed inter-day precision and accuracy of 2.5 to 3.6% C.V. and -0.9 to -0.4% R.E. Recovery of methocarbamol was 91.4-100.3% in five different lots of plasma. The method was shown to be applicable on different brands of C18 columns.
建立了一种等度高效液相色谱法并进行验证，用于定量测定人血浆中甲氯芬那酸的含量。取200微升人血浆中的甲氯芬那酸和内标，用乙酸乙酯萃取，蒸发至干，再用水复溶。采用反相C18柱，以甲醇-0.1M磷酸二氢钾-水（35:10:55，v/v/v）为流动相进行分离。检测波长为272nm的紫外光。线性范围为1-100微克/毫升（r>0.999）。定量限设定为1微克/毫升，以适合药代动力学研究。校准标准品的日间精密度和准确度的变异系数（C.V.）为1.0至3.6%，相对误差（R.E.）为-2.0至+1.6%。3、20和70微克/毫升的质量控制样品的日间精密度和准确度的C.V.为2.5至3.6%，R.E.为-0.9至-0.4%。甲氯芬那酸在五个不同批次的血浆中的回收率为91.4-100.3%。该方法适用于不同品牌的C18柱。
