以甲基橙为显色剂萃取光度法测定喜树中总生物碱
Extraction-Photometric Determination of Total Alkaloids in Camptotheca Acuminata with Methyl Orange as Color Reagent
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摘要: 为测定喜树中总生物碱含量,取样品的提取液100 μL,在pH 5的柠檬酸-磷酸盐缓冲介质中与甲基橙反应.反应混合液在25 ℃保持5 min后,用三氯甲烷10 mL进行萃取将生物碱转入有机相中,用光度法于366 nm波长处以三氯甲烷溶剂作参比测定其吸光度.用纯喜树碱作对照品,测得的吸光度与喜树碱质量浓度在6.5~104.0 mg·L-1范围内呈线性关系.取来自同一批的样品5份,按所提出的方法测定其总生物碱量,测定值的相对标准偏差为1.62%.在一已知样品的基础上,用标准加入法进行了回收试验,测得平均回收率为98.7%.Abstract: For determination of total alkaloids in camptotheca acuminata,100 μL of the extract of sample was reacted with methyl orange in a citrate-phosphate buffer medium of pH 5.The reaction mixture was kept at 25 ℃ for 5 min,and extracted with 10 mL of chloroform to transfer alkaloids into the organic phase,the absorbance of which was measured photometrically at 366 nm against chloroform as reference solution.Camptothecin was used as reference material in the determination.Linear relationship between values of absorbance and concentration of captothecin was obtained in the range of 6.5-104.0 mg·L-1.Precision was tested by analyzing 5 portions of samples from the same batch for their total alkaloid contents with the proposed method,value of RSD found was 1.62%.On the base of a known sample,test for recovery was made by standard addition method,value of average recovery obtained was 98.7%.