Idris Abubakr M, Elgorashe Rafea E E
King Khalid University, College of Science, Department of Chemistry, Abha, Saudi Arabia.
J AOAC Int. 2013 Mar-Apr;96(2):282-9. doi: 10.5740/jaoacint.11-057.
Sequential injection chromatography (SIC) is a recent, simple, and inexpensive green miniaturized separation technique. In the current study, SIC was exploited for the first time for biochemical analysis. A new SIC method for the separation and quantification of chlorpromazine in human urine, as well as pharmaceutical formulations, was developed and validated. Clozapine was used as an internal standard. Chlorpromazine was successfully separated on a C18 monolithic column (25x4.6 mm id). The UV detection was carried out at 250 nm using miniaturized fiber optic spectrometric devices. The optimum mobile phase composition was 30 mmol/L phosphate-acetonitrile-methanol (55.0 + 31.5 + 13.5, v/v/v) at pH 3.0. The sample volume was 40 microL and flow rate was 40 microL/s. Acceptable chromatographic results were obtained. The resolution was 2.7, peak symmetry was 1.1, and number of theoretical plates was more than 1 x 10(6). Good linearity (r = 0.9997) in the range of 25-100 microg/mL was also obtained. The method offered acceptable recovery for both human urine (89.6-93.1%) and pharmaceutical formulations (96.9-98.5%), which was sensitive enough to detect chlorpromazine. The LOD and LOQ in human urine were 61 and 204 ng/mL, respectively. The method was rapid and reagent-saving, and hence safe for the environment. The sample throughput was 26.3 samples/h and the total volume of consumed reagents was 4.0 mL.
顺序注射色谱法（SIC）是一种最新的、简单且廉价的绿色微型分离技术。在本研究中，首次将SIC用于生化分析。开发并验证了一种用于分离和定量人尿及药物制剂中氯丙嗪的新SIC方法。使用氯氮平作为内标。氯丙嗪在C18整体柱（内径25×4.6 mm）上成功分离。使用微型光纤光谱仪在250 nm处进行紫外检测。最佳流动相组成为30 mmol/L磷酸盐-乙腈-甲醇（55.0 + 31.5 + 13.5，v/v/v），pH值为3.0。进样体积为40 μL，流速为40 μL/s。获得了可接受的色谱结果。分离度为2.7，峰对称性为1.1，理论塔板数超过1×10⁶。在25 - 100 μg/mL范围内也获得了良好的线性关系（r = 0.9997）。该方法对人尿（89.6 - 93.1%）和药物制剂（96.9 - 98.5%）均提供了可接受的回收率，灵敏度足以检测氯丙嗪。人尿中的检测限和定量限分别为61和204 ng/mL。该方法快速且节省试剂，因此对环境安全。进样量为每小时26.3个样品，消耗试剂的总体积为4.0 mL。
