rochetbil和dbil项目介绍.ppt

Trans-esterification of either one or both acid function Structure of the unconjugated bilirubin Non polar molecule, internal H-bonding makes it only slightly water soluble - the free bilirubin is complexed to proteins ,this makes it more soluble - Non-covalent bond to ALB is needed the allow the transport of bilirubin in the blood DELTA bilirubin all the conjugated bilirubins mBc + dBC + deltaBc can be excreted in the bile * * * * TBIL slide is based on Diazo reaction. Dyphylline = accelerator Bu, Bc and Bd react to form azobilirubin Buffer layer - azobilirubin is pink in colour 540nm - pink colour 460nm - ‘blank’ for Hgb interference Dry Slide Technology Spreading Layer (BaSO4) TBIL Slide dyphylline diazonium salt Support Layer Buffer Layer buffer pH 3.0 [ T BIL ] = [ Bu ] +[ Bphoto] + [ mBc ] +[ dBc] +[ Bdelta] ALL BILIRUBIN FRACTIONS MESURED ON TBIL slide Tbil slide is based on Diazo réaction dyphilline = accelerator Bu , BC and delta bili are cplexed to azobilirubine Because of the buffer layer at pH=3 the Azo cplxe develop a pink color The slide is read at 540 nm = pink azo cplxe at 460 nM = eliminate the interference of the hemoglobine 干片法测定游离胆红素和葡萄糖醛酸化胆红素(Bu／Bc)： “BUBC”干片用于测定α，β和γ胆虹素。即游离胆红素和葡萄糖醛酸化的胆红素。单、双胆红素葡萄糖醛酸苷(BC)与媒染剂反应后反射光波长420-425 nm，而游离胆红素(Ru)最大反射波长在46O～465nm．这里不加重氮试剂，而直接利用通过媒染剂增强了的胆红素的内源色素媒染剂可使游离胆红素的反射强度增大两倍，对结合胆红素的效果就没有这么好。所幸的是，BU 和BC在460nm处的反射光谱可以完全分离，因而可 在一张干片上同时检测Bu 和BC。由于δ胆红素分于体积较大而被阻止在扩散层而无法到达反射层。所 BUBC干片不能测定δ胆红素。 当TBIL和BU/BC：都测定出来后。通过计算就能得到δ胆红素。因为： TBII=α+β+γ+δ胆红素 BUBC= α+β+γ胆红素 δ胆红素＝TBII－BUBC胆红素 All bilirubin fractions absorbs naturally at wavelengths between 400 and 540 ( remember bilirubins are yellow-orange pigments) the absorbance characteristics of BU and BC are enhanced by complexing the BC and BU to a substance called mordant . Complexing to mordant shifts BU and BC to 2 suitable wavelengths 400 and 460 nm at which the proportional s