North J A, Rein D, Tappel A L
Department of Food Science and Technology, University of California, Davis 95616, USA.
Anal Biochem. 1996 Jan 1;233(1):115-23. doi: 10.1006/abio.1996.0015.
Absorption spectra of heme proteins in a tissue homogenate contain information about the oxidation status of the biological sample. Deconvolution of absorption spectra can be used to quantitate the amount of individual heme proteins present in the mixture. The heme spectral analysis program (HSAP), a computer spreadsheet program, was used to quantitatively calculate values for heme proteins measured spectrally (390 to 450 and 500 to 640 nm) in tissue homogenates undergoing oxidation. The amount of oxidized heme proteins obtained by HSAP can be compared to other measurements of tissue oxidation. Precise quantitation of the amount of heme proteins present in a homogenate sample provided accurate assessment of the oxidized heme proteins calculated by HSAP. This quantitation was achieved through modification of existing pyridine hemochrome methods. Input into HSAP of the total heme protein content via the pyridine hemochrome value generated reproducible values for oxidized heme proteins. The program has broad potential as a multicomponent analysis tool. Modification of HSAP led to the development of a difference spectra analysis program (DSAP) which was used to quantitate the type and amount of heme proteins observed in mitochondrial difference spectra. In the present application, HSAP and DSAP provide methods for interpreting complex spectral information of multicomponent biological samples that undergo oxidation.
组织匀浆中血红素蛋白的吸收光谱包含有关生物样品氧化状态的信息。吸收光谱的反褶积可用于定量混合物中存在的单个血红素蛋白的量。血红素光谱分析程序（HSAP），一种计算机电子表格程序，用于定量计算在经历氧化的组织匀浆中通过光谱测量（390至450纳米和500至640纳米）的血红素蛋白的值。通过HSAP获得的氧化血红素蛋白的量可与组织氧化的其他测量值进行比较。对匀浆样品中存在的血红素蛋白量进行精确的定量分析，可对HSAP计算出的氧化血红素蛋白进行准确评估。这种定量是通过对现有的吡啶血红素方法进行改进实现的。通过吡啶血红素值将总血红素蛋白含量输入HSAP，可生成氧化血红素蛋白的可重复值。该程序作为一种多组分分析工具具有广泛的潜力。对HSAP的改进导致了差示光谱分析程序（DSAP）的开发，该程序用于定量线粒体差示光谱中观察到的血红素蛋白的类型和量。在本应用中，HSAP和DSAP提供了解释经历氧化的多组分生物样品复杂光谱信息的方法。
