Ceppa F, El Jahiri Y, Mayaudon H, Dupuy O, Burnat P
Laboratoire de Biochimie, Toxicologie et Pharmacologie cliniques, Hôpital d'Instruction des Armées Bégin, Saint-Mandé, France.
J Chromatogr B Biomed Sci Appl. 2000 Sep 15;746(2):115-22. doi: 10.1016/s0378-4347(00)00306-6.
A simple procedure for the determination of cotinine, major metabolite of nicotine in urine, is described. The assay involved a liquid-liquid extraction with dichloromethane in alkaline environment. The extract was dried at ambient temperature under a gentle stream of nitrogen. The residue was dissolved in 300 microl of mobile phase and 30 microl aliquot was injected via an automatic sampler into the liquid chromatograph and eluted with the mobile phase (10-9%, v/v methanol and acetonitrile, respectively in potassium dihydrogenphosphate buffer adjusted to pH 3.4) at a flow rate of 1 ml/min on a C8 Symmetry cartridge column (5 microm, 150 mm x 3.9 mm, Waters) at 25 degrees C. The eluate was detected at 260 nm. Internal standard was 2-phenylimidazole. Sensitive and specific, this technique was performed to test urine of diabetic patients (smokers and non-smokers) admitted in an endocrinology service. Urinary cotinine seems to be a better marker of smoking status than thiocyanates.
本文描述了一种用于测定尿液中可替宁（尼古丁的主要代谢产物）的简单方法。该分析方法包括在碱性环境中用二氯甲烷进行液 - 液萃取。萃取液在室温下通过缓慢的氮气流吹干。残余物溶解于300微升流动相中，取30微升等分试样通过自动进样器注入液相色谱仪，在C8 Symmetry柱（5微米，150毫米×3.9毫米，沃特世公司）上，于25℃下以1毫升/分钟的流速用流动相（分别为10 - 9%，v/v的甲醇和乙腈，在磷酸二氢钾缓冲液中，pH调至3.4）洗脱。洗脱液在260纳米处进行检测。内标物为2 - 苯基咪唑。该技术灵敏且特异，用于检测内分泌科收治的糖尿病患者（吸烟者和非吸烟者）的尿液。尿中可替宁似乎是比硫氰酸盐更好的吸烟状态标志物。
