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在强碱性水溶液中(pH>12),钙黄绿素(Calcein)与Ca~(2+)的络合物具有荧光性质(λ_(ex)=490nm,λ_(em)=520nm),而钙黄绿素本身在此条件下不发荧光。本文根据这一性质,在自制的多功能光导纤维光度计上建立了血清中钙的光导纤维荧光滴定法。方法以钙黄绿素为指示剂,以EDTA为滴定剂,根据滴定过程中试液的荧光强度与滴加EDTA标准溶液体积的关系曲线——滴定曲线确定滴定终点。此法取样量为50μL,甚至可少至1μL;精密度高,相对标准偏差小于2％;选择性好,血清中各种电解质和蛋白质都不干扰测定。用于测定临床血清试样中钙含量,结果令人满意。
The complex of calcein and Ca 2+ has the fluorescence property (λ ex (ex) = 490 nm and λ em (= 520 nm)) in strong alkaline aqueous solution (pH> 12) Itself does not fluoresce under this condition. Based on this property, this paper established a fluorescence spectrophotometric titration method of calcium in serum with a homemade multifunctional optical fiber photometer. Methods Calcein as indicator and EDTA as titrant were used to determine the end point of titration according to the curve of the titration curve between the fluorescence intensity of the test solution during titration and the volume of EDTA standard solution. This method sampling volume of 50μL, and even as little as 1μL; high precision, the relative standard deviation of less than 2%; good selectivity, serum electrolytes and proteins do not interfere with the determination. For the determination of calcium content in clinical serum samples, the results were satisfactory.