偏最小二乘-导数同步荧光法同时测定生物样品中对羟苯基乳酸、多巴和尿黑酸

在pH=6.00的KH2PO4-NaOH缓冲溶液中,波长差△λ=35nm,采集260～325nm波长范围内的同步荧光光谱数据,并对该数据进行一阶求导后用偏最小二乘法处理,以此建立了偏最小二乘-导数同步荧光光谱法同时测定对羟苯基乳酸(PHPLA)、多巴(Dopa)和尿黑酸(HGA)的新方法。该三种物质的线性范围分别为0.06～3.00、0.06～3.00和0.08～3.00mg/L;检出限分别为0.0307、0.0310和0.0428mg/L。该法用于细胞培养液中PHPLA、Dopa和HGA三组分的同时测定,其测定结果与高效液相色谱法的测定结果相比无显著性差异。

Simultaneous Determination of p-Hydroxyphenyllactic Acid, 3,4-Dihydroxyphenylalanine and 2,5-Dihydroxyphenylacetic Acid in Biological Sample by Derivative Synchronous Fluorescence Spectrometry Using Partial Least-squares Calibration

A simple,rapid and sensitive method was developed for simultaneous determination of phydroxyphenyllactic acid(PHPLA),3,4-dihydroxyphenylalanine(Dopa)and 2,5-dihydroxyphenylacetic acid(HGA)in this paper.Synchronous fluorescence(SFS)of PHPLA,Dopa and HGA were measured with wavelength interval of 35nm in KH2PO4-NaOH(pH=6.00)buffer medium.The experimental conditions were investigated and the spectra from 260to 325nm were tested,which were processed by derivative partial least square(DPLS).The linear ranges for PHPLA,Dopa and HGA were 0.06～3.00 mg/L,0.06～3.00mg/L and 0.08～3.00mg/L,respectively.The limits of detection for PHPLA,Dopa and HGA were 0.0307,0.0310and 0.0428 mg/L,respectively.The present method was successfully applied to simultaneous determination of PHPLA,Dopa and HGA in culture fluid and there was no significant difference between the results obtained by SFS-DPLS and high performance liquid chromatography.