Moriyama M, Yamashita S, Domoto H, Furuno K, Araki H, Gomita Y
Department of Hospital Pharmacy, Okayama University Medical School, Japan.
J Chromatogr B Biomed Sci Appl. 1999 Feb 19;723(1-2):301-5. doi: 10.1016/s0378-4347(98)00534-9.
Plasma phenobarbital (PB) concentrations in rat offspring were determined using a 9 microl capillary by high-performance liquid chromatography (HPLC). Capillary plasma which was put into a Bond Elut cartridge column by using 1 ml of 0.01 M KH2PO4 was applied to the column with 50 microl of 2 microg/ml of acetanilide (internal standard, I.S.). After washing the column, PB and I.S. were eluted with methanol and injected into the HPLC system. There were excellent linear correlation between the amount of PB and length of the capillary at three different concentrations. Calibration for PB was linear in the range of 0-50 microg/ml. The coefficients of variation were 3.4-5.0% and 5.9-7.5% in the within-day and between-day assays, respectively. The extraction recovery rates were 87.5-105.4%. By this method, it was possible to measure plasma PB concentrations in rat offspring without killing. These results suggested that this method is very useful to determine the plasma PB concentration derived from mother's milk in newborn rats.
采用9微升毛细管通过高效液相色谱法（HPLC）测定大鼠后代血浆中的苯巴比妥（PB）浓度。将1毫升0.01 M KH2PO4加入Bond Elut柱中，取50微升2微克/毫升的乙酰苯胺（内标，I.S.）加入其中，制成毛细管血浆并应用于该柱。柱清洗后，用甲醇洗脱PB和I.S.，并注入HPLC系统。在三种不同浓度下，PB含量与毛细管长度之间存在良好的线性相关性。PB的校准在0 - 50微克/毫升范围内呈线性。日内和日间测定的变异系数分别为3.4 - 5.0%和5.9 - 7.5%。提取回收率为87.5 - 105.4%。通过这种方法，可以在不处死大鼠的情况下测量其后代血浆中的PB浓度。这些结果表明，该方法对于测定新生大鼠母乳来源的血浆PB浓度非常有用。
