研究了二氢卟吩e6-C15单甲酯在不同溶剂体系中的荧光光谱特性, 据此建立了检测血浆中二氢卟吩e6-C15单甲酯的荧光分析法。 首先, 通过比较分析二氢卟吩e6-C15单甲酯在甲醇、 乙醇、 丙酮、 乙腈、 磷酸盐缓冲液和水六种不同溶剂体系中的荧光光谱特征, 考察了溶剂体系对二氢卟吩e6-C15单甲酯荧光光谱特性的影响, 结果发现甲醇和乙腈是比较理想的溶剂体系, 而磷酸盐缓冲液优于水溶液。 通过进一步考察有机溶剂含量和溶液pH对二氢卟吩e6-C15单甲酯发射波长和荧光强度的影响, 结果表明pH 7.2 磷酸盐缓冲液-乙腈(3∶7)混合溶液中二氢卟吩e6-C15单甲酯荧光强度大、 性质稳定。 采用pH 7.2 磷酸盐缓冲液-乙腈(3∶7)混合溶液沉淀SD大鼠血浆蛋白, 选择498.00和664.05 nm作为激发波长和发射波长, 建立的SD大鼠血浆样品中二氢卟吩e6-C15单甲酯荧光分析法专属性好, 灵敏度高, 线性范围为0.50～50.00 μg·mL-1(R2≥ 0.999), 批内和批间精密度RSD均小于10%, 提取回收率大于90%。 该方法简便、 快速、 可靠, 成功地应用于二氢卟吩e6-C15单甲酯在SD大鼠体内的药代动力学研究。

This paper studied the fluorescence spectral characteristic of chlorin e6-C15-monomethyl ester in different solvents to develop a fluorescence spectrometry for determining the concentration of chlorin e6-C15-monomethyl ester in plasma. By comparing the fluorescence spectral characteristics of chlorin e6-C15-monomethyl ester in six different solvents including methanol, ethanol, acetone, acetonitrile, phosphate buffered saline and water, the influence of different solvents on the fluorescence spectral characteristic of chlorin e6-C15-monomethyl ester has been examined. The results indicated that methanol and acetonitrile are the ideal solvent system, and the effect of phosphate buffered saline is better than water. The effect of organic solvent content and solution pH value on emission wavelength and fluorescence intensity was further evaluated. It was found that the fluorescence intensity of chlorin e6-C15-monomethyl ester was strong and stable in pH 7.2 phosphate buffered saline -acetonitrile (3∶7) solution and can be detected at the excitation wavelength of 498.00 nm and the emission wavelength of 664.05 nm. Based on this, we developed the fluorescence spectrometry for determining chlorin e6-C15-monomethyl ester. It is specific and sensitive with good linearity over the range of 0.5～50 μg·mL-1. The intra-batch and inter-batch precisions (RSD) were less than 10%, the extraction recoveries were all over 90%. The established method in this paper was simple, fast, effective and could be successfully applied to study the pharmacokinetics of chlorine e6-C15-monomethyl ester in SD rats.