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目的:定量测定长春花组织培养物中的吲哚生物碱含量。方法:用反相高效液相色谱法和紫外检测器测定了长春花组织培养物中吲哚生物碱的含量。分析柱:Nucleosil5C18柱,流动相:甲醇—乙腈—0025mol·L-1乙酸铵—三乙胺(15∶40∶45∶01),检测波长:280nm,流速:1mL·min-1。结果:色胺、蛇根碱、阿吗碱、文多灵和长春质碱的回收率分别是945%,959%,960%,970%和982%;平均日内精密度RSD均小于2%,日间精密度RSD均小于3%。结论:此法可有效分离和测定长春花组织培养物中吲哚生物碱含量,为进行细胞工程生产长春花生物碱提供了准确、灵敏、可靠的分析方法。
Objective: To quantitatively determine the content of indole alkaloids in Catharanthus roseus tissue culture. Methods: The contents of indole alkaloids in tissue culture of Catharanthus roseus were determined by RP-HPLC and UV detector. Analytical column: Nucleosil 5C18 column, mobile phase: methanol-acetonitrile-0.025 mol·L-1 ammonium acetate-triethylamine (15:40:45:01), detection wavelength: 280 nm, flow rate: 1 mL·min-1 . Results: The recoveries of tryptamine, serpentine, apoline, vindoline and catharanthine were 94.5%, 95.9%, 96.0%, 97.0% and 98.0%, respectively. The average intraday precision RSD was less than 2%, and the daytime precision RSD was less than 3%. Conclusion: This method can effectively isolate and determine the content of indole alkaloids in the tissue culture of Catharanthus roseus, providing an accurate, sensitive and reliable analytical method for cell engineering production of Vinca alkaloids.