Wang Li-ping, Wei Yong-ju
College of Chemistry and Material Science, Hebei Normal University, Shijiazhuang, China.
Yao Xue Xue Bao. 2012 Oct;47(10):1370-4.
Three-dimensional (3D) fluorescence spectra and thin layer fluorescence chromatogram of common Camptotheca fruit (CCF) crude drug, camptothecin (CPT) and 10-hydroxycamptothecin (HCPT) have been studied, and a novel fluorimetric method for determination of CPT in CCF crude drug has been established. In 3D fluorescence spectra, CPT presented 3 fluorescence peaks with excitation wavelengths lambdaex of 215, 255 and 365 nm, separately, and all peaks with emission wavelength lambdaem of 430 nm. HCPT presented 4 fluorescence peaks with lambdaex of 220, 265, 325 and 375 nm, separately, and all peaks with lambdaem of 555 nm. The fluorescence of CPT is much stronger than that of HCPT. Comparison of 3D fluorescence spectra and analysis of thin layer fluorescence chromatogram revealed that the main fluorescent component of CCF is CPT. HCPT and other components basically do not interfere with the fluorescence of CPT. Under the condition of pH 3.0-6.7, CCF aqueous solution can produce strong and steady fluorescence. Using methanol as solvent, the extracting solution of CCF was prepared, and diluted properly with water, then measured fluorescence intensity at lambdaex/lambdaem = 365/430 nm to determine the content of CPT. A linear calibration curve covered the concentration range 0.002 35-0.235 microg x mL(-1). The regression equation was IF = 9 + 30,844 c, with correlation coefficient r = 0.999 (n=9). The method has been applied to the analysis of CPT in a CCF sample, with a result of 0.127% and a spiked recovery rate of 102%. The reliability of the method has been verified by a thin layer chromatography-fluorescence scanning method. Experimental results demonstrated that this method can be used for quality evaluation of CCF crude drug.
研究了喜树果(CCF)药材、喜树碱(CPT)和10 - 羟基喜树碱(HCPT)的三维(3D)荧光光谱和薄层荧光色谱图,建立了一种测定CCF药材中CPT的新型荧光分析法。在3D荧光光谱中,CPT分别在激发波长λex为215、255和365 nm处呈现3个荧光峰,所有峰的发射波长λem均为430 nm。HCPT分别在λex为220、265、325和375 nm处呈现4个荧光峰,所有峰的λem均为555 nm。CPT的荧光比HCPT的荧光强得多。3D荧光光谱的比较和薄层荧光色谱图分析表明,CCF的主要荧光成分是CPT。HCPT和其他成分基本不干扰CPT的荧光。在pH 3.0 - 6.7的条件下,CCF水溶液能产生强而稳定的荧光。以甲醇为溶剂制备CCF提取液,用水适当稀释,然后在λex/λem = 365/430 nm处测量荧光强度以测定CPT的含量。线性校准曲线覆盖的浓度范围为0.002 35 - 0.235 μg·mL⁻¹。回归方程为IF = 9 + 30844c,相关系数r = 0.999(n = 9)。该方法已应用于CCF样品中CPT的分析,结果为0.127%,加标回收率为102%。该方法的可靠性已通过薄层色谱 - 荧光扫描法验证。实验结果表明,该方法可用于CCF药材的质量评价。