高效液相色谱法对血红素加氧酶的微量测定：应用于人体肝脏穿刺活检样本的测定
Microassay of heme oxygenase by high-performance liquid chromatography: application to assay of needle biopsies of human liver.
作者信息
Lincoln B C, Mayer A, Bonkovsky H L
机构信息
Department of Medicine, Emory University, Atlanta, Georgia 30322.
出版信息
Anal Biochem. 1988 May 1;170(2):485-90. doi: 10.1016/0003-2697(88)90662-8.
We developed a microassay for heme oxygenase, in which bilirubin (BR) production was measured by HPLC, and compared it to previously reported spectrophotometric methods. The microassay required as little as 5 mg wet human, rat, or chick embryo liver. Using the HPLC assay, values for heme oxygenase activity in extracts (10,000 g supernatant) of normal human liver obtained by needle biopsies were 44 +/- 7 (pmol BR.min-1.mg protein-1). Spectrophotometric assays of homogenates of human liver resulted in low values for heme oxygenase, due to unknown sources of interference. Comparative values of microsomal heme oxygenase activity were 294 +/- 25, 95 +/- 3, and 87 +/- 9 pmol BR.min-1.mg protein-1 for chick, rat, and human livers, respectively.
我们开发了一种用于血红素加氧酶的微量测定法，其中胆红素（BR）的产生通过高效液相色谱法（HPLC）进行测定，并将其与先前报道的分光光度法进行比较。该微量测定法所需的湿人、大鼠或鸡胚肝脏组织量低至5毫克。使用HPLC测定法，通过针吸活检获得的正常人肝脏提取物（10,000g上清液）中的血红素加氧酶活性值为44±7（pmol BR·min⁻¹·mg蛋白质⁻¹）。由于未知的干扰源，对人肝脏匀浆进行分光光度测定得到的血红素加氧酶值较低。鸡、大鼠和人肝脏微粒体血红素加氧酶活性的比较值分别为294±25、95±3和87±9 pmol BR·min⁻¹·mg蛋白质⁻¹。
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