摘要
目的:建立高效液相色谱定量分析虫草制品中8种核苷及碱基的方法。方法:用 HPLC 测定虫草制品超声提取液中的尿嘧啶、胞苷、次黄嘌呤、尿苷、腺嘌呤、鸟苷、腺苷和3’-脱氧腺苷含量。色谱条件:Kromasil C_8柱(200 mm×4.6 mm,5μm),采用梯度洗脱,流动相甲醇-双蒸水(2:98)10 min(15:85)25 min,流速1 mL·min^(-1),柱温30℃,检测波长260 nm,进样量20μL,灵敏度0.01 AUFS。结果:此法成功应用于不同来源虫草制品中核苷及碱基的测定,在相应浓度范围内各组分线性相关系数均高于0.9988,精密度和重复性试验的 RSD 值都小于5.0%,8种组分的回收率为95.8%～102.5%,含量8 h内稳定。实验表明不同菌种、培养方法所得虫草产品及同一发酵产物的不同部位其核苷、碱基含量有较大差异。结论:本测定方法快速、简便,能用于不同来源虫草及其发酵品中多种核苷、碱基的同时测定,对于全面评估虫草制品的质量有重要意义。
Objective： To developed a method with HPLC to simultaneously determine nucleosides and bases in Cordyceps sp. fermentation preparations. Methods： HPLC was adopted to measure the amount of uracil, cytidine, hypoxanthine ,uridine, adenine, guanosine, adenosine and 3＇-deoxyadenosine in Cordyceps sp. fermentation preparations. The working conditions were ： column, Kromasil C8 （200 mm × 4. 6 mm,5 μm） ; solvent A, double distilled water; solvent B, methanol; linear gradient ： methanol - water （ 2 ： 98 ） 10 min ^0.2min→ （ 15 ： 85 ） 25 min; flow rate, 1.0 mL · min^-1 ;temperature,30℃ ;detective wavelength,260 nm. Results：The method was successfully applied to analyze nucleosides and bases in Cordyceps sp. fermentation preparations from various origins. The correlation coefficients of the standard calibration curves of eight components were all higher than 0. 9988 over the corresponding concentration range,the RSD of the accuracy and reproducibility of the assay were all less than 5.0% ; the mean recoveries of these eight components were 95.8% - 102. 5%, and components were stable within 8 h. The experiment showed the significant difference in quantity of nueleosides and bases from various oringins as well as the different accumulation regions in the Cordyceps militaris fermentation preparations formed by some of the nueleosides and bases. The results also indicated that under the similar culture conditions ,the submerged and solid substrate fermentation preparations of the same Cordyceps sinensis strain produced different amounts of nueleosides and bases. Conclusion： The constructed HPLC method is simple, fast and convenient,it is suitable for assay of several nueleosides and bases of Cordyceps sp. fermentation preparations. Thus, it is valuable for evaluating the quality of productions comprehensively.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2006年第7期953-957,共5页
Chinese Journal of Pharmaceutical Analysis
基金
上海市重点学科建设项目资助
关键词
高效液相色谱
冬虫夏草
蛹虫草
核苷
碱基
发酵制品
HPLC
Cordyceps sinensis
Cordyceps militaris
nucleosides
bases
fermentation preparations
