RP-HPLC进行甲氨蝶呤血药浓度监测时保留时间校准方法研究

目的 建立RP—HPLC进行MTX血药浓度监测时保留时间校准方法。方法应用容量因子与流动相比例的三次方多项式模型，建立MTX流动相比例与保留时间的函数，并通过此函数用保留时间求算流动相比例，与理论流动相比例比较，从而计算流动相应该补充组分的量，达到调整保留时间的目的。结果四种不同色谱条件下容量因子与流动相比例的三次方多项式模型拟合关系良好，R0均为1．000，保留时间经校准后RSD为0.9％(n=5)。结论本方法能准确校准因流动相比例引起变化的保留时间，适用于MTX血药浓度监测时保留时间的校准，方法新颖、有效、简单。

RP-HPLC method for calibrating the error of retention time of MTX

OBJECTIVE To establish the method for calibrating the error of the retention time of MTX by RP-HPLC.METHODS We established a function between the mobile phase ratio of MTX and the retention time by cubic model of capacity factor and mobile phase ratio. Using the function the mobile phase ratio was calculated by the retention time. Then the result was compared with theoretical mobile phase ratio to determine how much mobile phase should be complemented. RESULTS The relation between the capacity factor and the mobile phase ratio matched the cubic model under 4 chromato-graph conditions. The correlation was 1.00. The RSD of retention time that had been calibrated was 0.9% (n = 5). CONCLUSIONS The method could calibrate the retention time error caused by mobile phase ratio, which suits the retention time calibration in MTX determination in serum by RP-HPLC. It is novel, effective and simple.