Koch J P, ten Tusscher G W, Koppe J G, Guchelaar H J
University of Utrecht, Faculty of Pharmacy, Department of Analysis and Toxicology, The Netherlands.
Biomed Chromatogr. 1999 Jun;13(4):309-14. doi: 10.1002/(SICI)1099-0801(199906)13:4<309::AID-BMC881>3.0.CO;2-J.
In this study the validation of a reversed-phase high-performance liquid chromatography (HPLC) method, with UV-detection, for both caffeine and paraxanthine in human serum is described. This method is feasible for cytochrome P450 1A2 (CYP1A2) phenotyping, according to the results of a pilot study. With this HPLC method caffeine and paraxanthine can be determined selectively and specifically. In the expected concentration range, caffeine recoveries were 98-108% (within-run variation 4.0-6.4%, between-run variation 6.4-8.8%), paraxanthine recoveries were 96.6-97.5% (within-run variation 5.0-7.2%, between-run variation 7.2-10.8%). The limits of detection for caffeine and paraxanthine using this HPLC system were 0.3 and 0.1 mg/L, respectively. Linear calibration curves for both caffeine and paraxanthine were obtained in the concentration range 0.5-30 mg/L (r > 0.9999. Serum samples were stable for a week, when stored at -20 and +4 degrees C.
本研究描述了一种用于检测人血清中咖啡因和副黄嘌呤的反相高效液相色谱（HPLC）紫外检测方法的验证。根据初步研究结果，该方法对于细胞色素P450 1A2（CYP1A2）表型分析是可行的。使用这种HPLC方法，可以选择性且特异性地测定咖啡因和副黄嘌呤。在预期浓度范围内，咖啡因回收率为98 - 108%（批内变异4.0 - 6.4%，批间变异6.4 - 8.8%），副黄嘌呤回收率为96.6 - 97.5%（批内变异5.0 - 7.2%，批间变异7.2 - 10.8%）。使用该HPLC系统时，咖啡因和副黄嘌呤的检测限分别为0.3和0.1 mg/L。咖啡因和副黄嘌呤在0.5 - 30 mg/L浓度范围内均获得了线性校准曲线（r > 0.9999）。血清样本在-20℃和+4℃下储存时，可稳定保存一周。
