Mizobe M, Kondo F, Kumamoto K, Terada T, Nasu H
Department of Veterinary Public Health, Faculty of Agriculture, Miyazaki University, Japan.
Microbios. 1996;86(346):39-47.
Rapid and quantitative analytical methods for bilirubin using high-performance liquid chromatography (HPLC) with UV detection were developed for samples from equines at a meat inspection site. Sharp HPLC peaks for bilirubins, unconjugated bilirubin (UCBL) and conjugated bilirubin (CBL), were obtained using a simple mobile phase of methanol:0.5 M Tris-HCl buffer (65:35, v/v, pH 7.4). A variable wavelength detector set at 450 nm, 0.01 AUFS and a recorder set at 4 cm/min were used for detection. Peaks for UCBL and CBL occurred at 7.1 min and 4.9 min, the lower limits of detection ranged between 0.16 and 0.78 microgram/ml, respectively. Orange II, for which the retention time was 3.6 min, was selected as an internal standard. When the samples were analysed from healthy equines and those suspected of being jaundiced due to a yellow colour change in the carcasses, only UCBL peaks were recognized, and CBL peaks were never obtained. Aqueous humour is a very common sample at meat inspections but UCBL or CBL peaks were all absent in both healthy and clinical samples from equines. There were reliable correlations between UCBL peaks (microV) using HPLC and total bilirubin value (mg/dl) measured by the total bilirubin assay kit in serum. A correlation between peak height (microV) on HPLC from serum samples and that of pericardial fluid samples was obtained.
针对肉类检验场所的马类样本，开发了采用高效液相色谱（HPLC）结合紫外检测的胆红素快速定量分析方法。使用甲醇：0.5M Tris - HCl缓冲液（65:35，v/v，pH 7.4）的简单流动相，获得了胆红素、未结合胆红素（UCBL）和结合胆红素（CBL）的尖锐HPLC峰。使用设定在450nm、0.01 AUFS的可变波长检测器和设定在4cm/min的记录仪进行检测。UCBL和CBL的峰分别出现在7.1分钟和4.9分钟，检测下限分别在0.16至0.78微克/毫升之间。选择保留时间为3.6分钟的橙黄II作为内标。当对健康马类以及因胴体颜色变黄而疑似黄疸的马类样本进行分析时，仅识别出UCBL峰，从未获得CBL峰。房水是肉类检验中非常常见的样本，但在健康和临床马类样本中均未出现UCBL或CBL峰。HPLC测定的UCBL峰（微伏）与血清中总胆红素检测试剂盒测定的总胆红素值（mg/dl）之间存在可靠的相关性。获得了血清样本和心包液样本在HPLC上的峰高（微伏）之间的相关性。
