反相高效液相色谱法测定长春花组织培养物中吲哚生物碱含量

赵剑; 朱蔚华; 吴蕴祺; 胡秋

反相高效液相色谱法测定长春花组织培养物中吲哚生物碱含量

DETERNINATION OF INDOLE ALKALOIDS IN CATHARANTHUSROSEUS TISSUE AND CELL CULTURES BY RPHPLC

摘要

摘要: 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—0.025mol·L^-1乙酸铵—三乙胺(15∶40∶45∶0.1),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94.5%,95.9%,96.0%,97.0%和98.2%;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。

Abstract: AIM: To determine the indole alkaloids tryptamine, serpentine, vindoline, ajmalicine and cathananthine in Catharanthus roseus tissue and cell cultures by RPHPLC and UV detector. METHODS: These indole alkaloids were successfully seperated on a Nucleosil 5 C₁₈ column (250 mm×4.6 mm, 5 μm) with a mobile phase of methanol — acetonitrile — 0.025 mol·L^-1 ammonium acetate — triethylamine (15∶40∶45∶0.1) by UV detector at 280 nm. RESULTS: The average recoveries and equations of linear regression of tryptamine, serpentine, vindoline, ajmalicine and catharanthine were 94.5%, 95.9%, 97.0%, 96% and 98.2%, respectively, and Y=85157143X-185275 (γ=0.9988), Y=43939499X-3414.24 (γ=0.9996), Y=10386.97X-357.53 (γ=0.9994), Y=40717.75X-1551.22 (γ=0.9996), Y=5694.16X-1668.667 (γ=0.9979), respectively. Intra and inter day precisions of these alkaloids were lower than 2% and 3% (RSD), respectively. CONCLUSION: A simple, rapid and sensitive method with good precision was established for the determination of these alkaloids in cell cultures of Catharanthus roseus.

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