Taulier A, Levillain P, Lemonnier A
Ann Biol Clin (Paris). 1986;44(3):242-8.
We are proposing a technique enabling the dosage of hemoglobin in plasma or urine using spectrophotometry in derivative double dash. Around 420 nm hemoglobin has an elevated molecular extinction factor but the absorption of natural pigments creates a non-negligible interference in the classic spectrophotometric methods. On the contrary, the use of the derivative double dash enables to practically eliminate the background noise and to obtain a specific signal for hemoglobin while keeping an excellent sensitivity. In the plasma, where hemoglobin is entirely linked to haptoglobin, the dosage is done by direct reading (while in the urine the presence of several pigment forms requires the use of Drabkin's reagent. The method is simple, quick, precise and enables to titrate 1 mg.l (or 0.015 mumole.l-1) of hemoglobin. The influence of bilirubin is negligible. The comparison with a method using Allen's correction shows the superiority of the former as far as precision and accuracy are concerned, by avoiding the danger of a non-negligible systematic error.
我们提出了一种利用导数双波长分光光度法测定血浆或尿液中血红蛋白含量的技术。在420纳米左右，血红蛋白具有较高的分子消光系数，但天然色素的吸收在经典分光光度法中会产生不可忽略的干扰。相反，使用导数双波长能够实际消除背景噪声，并在保持出色灵敏度的同时获得血红蛋白的特定信号。在血浆中，血红蛋白完全与触珠蛋白结合，通过直接读数进行测定（而在尿液中，由于存在多种色素形式，需要使用德拉布金试剂）。该方法简单、快速、精确，能够滴定1毫克/升（或0.015微摩尔/升）的血红蛋白。胆红素的影响可忽略不计。与使用艾伦校正的方法进行比较表明，就精密度和准确度而言，前者具有优势，因为它避免了不可忽略的系统误差的风险。
