薄层扫描测定中荧光与增荧光成分的最佳激发波长研究

Study of Best Excitation Wave Length of Fluorescence and Fluorescence Increase Components in Thin Slice Scan and Detection

目的:为排除干扰,提高荧光扫描的灵敏度,对荧光与增荧光成分的最佳激发波长进行了选择研究。方法:采用同一荧光斑点,以不同的滤光片和不连续的波长,逐一激发,记录其相应强度积分值,以波长为横坐标,积分值为纵坐标,绘制曲线图,取曲线图的最大积分值条件,作为最佳激发波长和首选滤光片。结果:通过对盐酸小檗碱、蛇床子素、丹皮酚与胆酸的优选,其最佳激发波长分别为(338±2)nm、(320±2)nm、(292±2)nm和(385±2)nm,首选滤光片分别是3号、1号、1号和2号,灵敏度高,点样量在ng级,即可准确定量。结论:证实了荧光化合物都沿用(366±1)nm为激发波长的不合理性,揭示了首选滤光片和激发波长因化合物不同而异,验证方法时,需通过优选。

Objective: in order to get rid of disturbance and improve the sensibility of fluorescent scanning,the best excitation wave length of fluorescence and fluorescence increase was selectively explored.Methods: With the same fluorescent spot,different spectral filter,discontinuous wave length and excitation in sequence,record its corresponding intensity score;draw the curve chart with the wave length as abscissa and the score as ordinate,taking the highest score in the chart as the best excitation wave length and the preferable spectral filter.Results: Through optimizing berberine hydrochloride,osthole,paeonol and colalin,the best excitation wave lengths were(338±2)nm,(320±2)nm,(292±2)nm and(385±2)nm,and the preferable filters were No.3,No.1 and No.2 with high sensibility,sample application at ng level,which could quantitate accurately.Conclusion: It is proved unreasonable that fluorescent chemicals continue to use(366±2)nm as excitation wave length,and revealed that the best filter and excitation wave length are different because of different chemicals.Optimization is preferred in verifying methods.