薄层扫描测定中荧光与增荧光成分的最佳激发波长研究

摘要

Abstr act 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 cor-responding 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 unrea-sonable 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.%　　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。