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目的:基于钙黄绿素-铜(Ⅱ)荧光体系测定乙酰半胱氨酸。方法:在pH=8.0的Na2HPO.412H2O-KH2PO4缓冲液中,以492 nm为激发波长,520 nm为发射波长测定乙酰半胱氨酸溶液的荧光强度。结果:在pH=8.0的Na2HPO.412H2O-KH2PO4缓冲液中,二价铜离子与钙黄绿素配位引起荧光猝灭。由于乙酰半胱氨酸中巯基上的硫离子与Cu2+的亲和力很强,可从钙黄绿素-铜(Ⅱ)的络合物中夺取铜离子而使钙黄绿素游离出来,从而使体系的荧光得以恢复,并且荧光恢复的程度与加入乙酰半胱氨酸的量在一定范围内成线性。结论:建立了一种测定乙酰半胱氨酸的荧光分析新方法,该方法的线性范围为6.0 10-6～1.4 10-5 mol/L,检出限为4.010-6 mol/L。
Objective: To determine acetylcysteine based on calcein - copper (Ⅱ) fluorescence system. Methods: The fluorescence intensity of acetylcysteine solution was measured at 492 nm as the excitation wavelength and 520 nm as the emission wavelength in Na2HPO.412H2O-KH2PO4 buffer solution with pH = 8.0. Results: In Na2HPO.412H2O-KH2PO4 buffer solution with pH = 8.0, the coordination of divalent copper ions with calcein caused fluorescence quenching. Since the affinity of sulfanilic acid on sulfhydryl in Cu2 + is very high, copper can be extracted from the complex of calcein-copper (Ⅱ) to release calcein, so that the fluorescence of the system can be restored , And the degree of fluorescence recovery and the amount of acetylcysteine added were linear within a certain range. Conclusion: A new fluorescence analysis method for the determination of acetylcysteine was developed. The linear range of this method is 6.0 10-6 ~ 1.4 10-5 mol / L with the detection limit of 4.010-6 mol / L.