【2017年整理】傅里叶变换红外光谱诊断地中海贫血症 3.docVIP

傅里叶变换红外光谱诊断地中海贫血症 【摘要】  为建立简单快速的地中海贫血诊断方法，本研究探讨了以傅里叶变换红外光谱结合水平衰减全反射（FTIRHATR）技术在地中海贫血诊断中的制样方法及光谱的数据处理方法。在制样预处理中，通过对样品进行稀释并干燥成膜消除水分子对光谱吸收干扰，保持ATR光谱中各波长对样品的穿透深度一致。结果表明，当1652 cm－１吸收度小于1.5时（即透射率T小于4%时），各波峰强度与血红蛋白浓度呈良好的线性关系（r0.995）及实验重复性（RSD4%）。在数据处理上，改进的相对强度方法用于800～1780 cm－１和2480～3600 cm－１区间的分析。通过与常规的傅里叶去卷积谱及差谱方法相比，本方法可消除样品浓度所带来的影响因素，灵敏地揭示群体数据中组分与结构在不同组间的显著差异，如1638 cm－１处重叠的蛋白二级结构峰，1172 cm－１、1440 cm－１表征脂类物质的吸收峰，1064 cm－１表征磷酸化合物峰位及表征SH的2553 cm－１附近的吸收峰在正常组与地中海贫血组间存在显著差异。从而避免了几个峰位的相对强度所反映的信息不足及选择参比峰的困扰，对揭示整体的差异变化规律有着重要的作用。 【关键词】 傅里叶变换红外光谱，水平衰减全反射，地中海贫血，相对强度 　　Identification of Thalassemia by Fourier Transform Infrared　Spectroscopy and Spectral Data Processing 　　PENG LiＸin, WANG GuiＷen, YAO HuiＬu, HUANG ShuＳhi, WANG YiＢing, TAO ZhanHua, LI YongＱing 　　1(Biophysics Laboratory, Guangxi Academy of Sciences, Nanning 530003) 　　2(Department of Physics, East Carolina University, Greenvile, NC 278584353, USA) 　　Abstract To set up a rapid, simple diagnosis of thalassemia, Fourier transform infrared spectroscopy (FTIR) associated with horizontal attenuated total reflectance (HATR) was firstly used to diagnosis βthalassemia patients. Here, the sample preprocessing and data processing were discussed in detail. Properly diluted samples were filmed on ATR crystal surface and then all evanescent waves can penetrate the sample completely. The IR spectra indicated excellent linearity (r0.995) and reproducibility (RSD4%) under 1.5 absorbance unit at 1652 cm－１. Modified intensity ratios method was used in 800－1780 cm－１ and 2480－3600 cm－１ regions respectively. It demonstrated protein secondary structure change near 1638 cm－１ and others significant differences such as bands at 1172 cm－１ and 1440 cm－１ that distribute to lipid, band at 1064 cm－１ arising from phosphate substance and 2553 cm－１ that originating from SH stretching vibration. This method not only avoids the problems of large number of free parameters comparison with other data processing but also can