Boutagy J, Shenfield G M
Department of Clinical Pharmacology, Royal North Shore Hospital, St. Leonards, N.S.W., Australia.
J Chromatogr. 1991 Apr 19;565(1-2):523-8. doi: 10.1016/0378-4347(91)80420-h.
A simple, specific and rapid reversed-phase high-performance liquid chromatographic (HPLC) procedure for sotalol determination is described requiring small plasma volumes. The high recovery of sotalol from plasma and the high precision of measurement obviate the need for an internal standard. Plasma samples (300 microliters) were deproteinised with 50 microliters of 70% (w/w) perchloric acid in disposable glass tubes. After vortex-mixing and centrifugation, 30 microliters of 4 M K2HPO4 were added followed by gentle shaking. A 20-microliters aliquot was then injected (by autosampler) for HPLC analysis. Chromatography was performed on a glass-lined 250 mm x 4 mm 5-micron C18 steel column. The mobile phase was 6% (v/v) acetonitrile in 0.08 M KH2PO4 buffer (pH 4.6). The flow-rate was 0.8 ml/min. Detection was by fluorescence with excitation and emission wavelengths at 235 and 310 nm, respectively. The retention time for sotalol was 7.1 min. Calibration was linear from 0.16 to 10 micrograms/ml in plasma (r greater than 0.999 for detector response to sotalol). The minimum concentration for quantitation was 0.08 micrograms/ml [within assay coefficient of variation (C.V.) less than 5%]. Recovery was near quantitative (greater than 98%) and replicate (intra-assay precision was less than 5% C.V.). Analysis of samples (n = 10) at concentrations of 0.42 and 4.2 micrograms/ml gave mean values of 0.44 and 4.3 micrograms/ml, respectively. The inter-assay C.V. values were 4.5 and 2.2%, respectively. Other clinically used antiarrhythmic drugs did not interfere. This assay can be performed using other commercial C18 analytical columns by suitable adjustment of mobile phase flow-rate and acetonitrile composition.
本文描述了一种简单、特异且快速的反相高效液相色谱（HPLC）法用于索他洛尔的测定，该方法所需血浆体积小。索他洛尔从血浆中的高回收率以及测量的高精度使得无需内标。在一次性玻璃管中，用50微升70%（w/w）的高氯酸对300微升血浆样品进行脱蛋白处理。涡旋混合并离心后，加入30微升4M的K2HPO4，然后轻轻振荡。接着（通过自动进样器）注入20微升等分试样进行HPLC分析。色谱分析在一根内衬玻璃的250mm×4mm、5微米的C18钢柱上进行。流动相为0.08M KH2PO4缓冲液（pH 4.6）中6%（v/v）的乙腈。流速为0.8ml/min。检测采用荧光法，激发波长和发射波长分别为235nm和310nm。索他洛尔的保留时间为7.1分钟。血浆中校准曲线在0.16至10微克/毫升范围内呈线性（检测器对索他洛尔响应的r大于0.999）。定量的最低浓度为0.08微克/毫升[批内变异系数（C.V.）小于5%]。回收率接近定量（大于98%）且重复性好（批内精密度小于5% C.V.）。对浓度为0.42和4.2微克/毫升的样品（n = 10）进行分析，平均值分别为0.44和4.3微克/毫升。批间C.V.值分别为4.5%和2.2%。其他临床使用的抗心律失常药物不产生干扰。通过适当调整流动相流速和乙腈组成，使用其他市售C18分析柱也可进行该测定。
