柱前衍生化高效液相色谱法测定蝉花菌丝体中多球壳菌素含量___

 

目的：建立柱前衍生化高效液相色谱法测定蝉花菌丝体中多球壳菌素含量。方法：样品采用甲醇超声提取，固相萃取小柱纯化，用邻苯二甲醛衍生化，高效液相色谱法测定。色谱柱为Eclipse XDB-C₈柱(150 mm×4.6 mm,5 μm)；流动相为乙腈-水(60∶40)，流速1.0 mL·min^-1，检测波长260 nm，柱温30 ℃。结果：多球壳菌素浓度在6.0~360.0 μg·mL^-1范围内与其衍生物峰面积呈良好的线性关系(r=0.9985)；平均加样回收率(n=6)为989%，RSD=2.9%。结论：方法选择性好，灵敏度高，结果准确、可靠，可用于测定蝉花菌丝体中多球壳菌素含量。

 

 

 

Objective:To develop a precolumn derivatization-HPLC method for the determination of myriocin in Cordyceps cicadae mycelia.Methods:Sample was extracted with methanol,cleaned up with solid phase extraction(SPE) and derivatized with o-phthalaldehyde.The separation was performed on an Eclipse XDB-C₈ column(150 mm×4.6 mm,5 μm);The mobile phase was composed of acetonitrile-water(60∶40) with the flow rate of 1.0 mL·min-1;The detection wavelength was 260 nm and the column temperature was 30 ℃.Results:A linearity between derivative peak area and myriocin concentration was achieved in the range of 6.0-360.0 μg·mL^-1(r=0.9985).The average recovery was 98.9%,RSD=2.9%.Conclusion:The method is accurate and reliable,and can be used for determination of myriocin in C.cicadae mycelia.

 1CHEN Wan-qun(陈万群),CHEN Gu-rong(陈古荣).Study development of the substitute of Cordyceps sinensis(冬虫夏草代用品研究进展).Chin Tradit Herb Drugs(中草药),1994,25(5):269

2HU Feng-lin(胡丰林),LI Zeng-zhi(李增智).Secondary metabolites and their bioactivities of Cordyceps and its related fungi(虫草及相关真菌的次生代谢产物及其活性).Mycosystema(菌物学报),2007,26(4):607

3Fujita T,Inoue K,Yamamoto S,et al.Fungal metabolites.Part 11.A potent immunosuppressive activity found in Isaria sinclairii metabolite. J Antibiot,1994,47(2):208

4Miyake Y,Kozutsumi Y,Nakamura S, et al.Serine palmitoyltransferase is the primary target of a sphingosine-like immunosuppressant,ISP-1/myriocin.Biochem Biophys Res Comm,1995,211(2):396

5Hojjati MR,Li ZQ,Zhou HW, et al.Effect of myriocin on plasma sphingolipid metabolism and atherosclerosis in apoE-deficient mice.J Biol Chem,2005,280(11):10284

6XIAO Zhao-hua(肖朝华),ZHOU Jian-hua(周建华),WU Heng–sheng(吴衡生).Effect of myriocin on hypertrophy of glomerular mesangial cells and cell cycle proteins(多球壳菌素对肾小球系膜细胞肥大及细胞周期蛋白的影响).Chin J Nephrol(中华肾脏病杂志),2006,22(8):507

7YU Jia-wen(余佳文),MO Zhi-hong(莫志宏),YANG Ming-jing(杨明静),et al.Determination of myriocin in Cordyceps cicadae mycelia by HPLC with solid-phase extraction(固相萃取结合HPLC测定蝉花菌丝体中多球壳菌素的含量).Chin Pharm J (中国药学杂志),2008,43(15):1191