采用在线柱后紫外照射和荧光检测的反相高效液相色谱法测定人血清中的舒林酸及其代谢物。

Determination of sulindac and its metabolites in human serum by reversed-phase high-performance liquid chromatography using on-line post-column ultraviolet irradiation and fluorescence detection.

作者信息

Siluveru M, Stewart J T

机构信息

Department of Medicinal Chemistry, College of Pharmacy, University of Georgia, Athens 30602-2352, USA.

出版信息

J Chromatogr B Biomed Appl. 1995 Nov 3;673(1):91-6. doi: 10.1016/0378-4347(95)00239-f.

DOI:10.1016/0378-4347(95)00239-f

PMID:8925079

Abstract

On irradiation with ultraviolet light, the antiinflammatory agent sulindac and its two metabolites sulindac sulfone and sulindac sulfide form highly fluorescent derivatives. This reaction was exploited for the sensitive and selective detection of these compounds in serum using reversed-phase high-performance liquid chromatography on a Ultrasphere octylsilane column (150 x 4.6 mm I.D.) at ambient temperature with a flow-rate of 0.5 ml/min. The analytes of interest were isolated from serum using a Bond-Elut C2 column with satisfactory recovery and selectivity. The detection limits were 10 ng/ml for each of the three analytes using 1 ml of serum and the limit of quantitation was 50 ng/ml. Linear calibration curves from 50 to 1000 ng/ml for all three analytes show coefficients of determination of 0.9999. The post-column ultraviolet irradiation was optimized and the effect of irradiation time on the fluorescence response was determined for all three analytes. Precision and accuracy of the method were 0.4-5.6 and 1.6-4.5% for sulindac, 2.3-5.6 and 1.4-5.3% for sulindac sulfone and 2.5-4.3 and 0.8-2.8% for sulindac sulfide, respectively.

摘要

在紫外线照射下，抗炎药舒林酸及其两种代谢产物舒林酸砜和舒林酸硫化物会形成高荧光衍生物。利用这一反应，采用反相高效液相色谱法，在室温下于Ultrasphere辛基硅烷柱（内径150×4.6 mm）上，以0.5 ml/min的流速，对血清中的这些化合物进行灵敏且选择性的检测。使用Bond-Elut C2柱从血清中分离出目标分析物，回收率和选择性均令人满意。使用1 ml血清时，三种分析物的检测限均为10 ng/ml，定量限为50 ng/ml。所有三种分析物在50至1000 ng/ml范围内的线性校准曲线的决定系数为0.9999。对柱后紫外线照射进行了优化，并确定了照射时间对所有三种分析物荧光响应的影响。该方法对舒林酸的精密度和准确度分别为0.4 - 5.6%和1.6 - 4.5%，对舒林酸砜分别为2.3 - 5.6%和1.4 - 5.3%，对舒林酸硫化物分别为2.5 - 4.3%和0.8 - 2.8%。