灵芝孢子粉中总三萜含量标准测定方法的改进  

Optimization of a Standard Protocol for the Quantitative Determination of Total Triterpenoids in Ganoderma lucidum

作　　者：王红梅[1] 刘海燕[1] 林淼[1] 雷萍[1] 金晓芬[1] 

机构地区：[1]上海市农业科学院农产品质量标准与检测技术研究所,上海201403

出　　处：《食用菌学报》2013年第2期42-45,共4页Acta Edulis Fungi

摘　　要：对农业部部颁备案灵芝（Ganodermalucidum）孢子粉总三萜含量测定标准NY／sJ339—2001进行优化，将齐墩果酸标准溶液浓度从1mg／mL优化为0．2mg／mL，取样体积范围从0．02～0．1mL优化为0．1～0．6mL，即样品质量范围从20～100Mg变化为20--120μg；取样后添加沸水浴加热挥去溶剂这一优化步骤；显色剂5％香草醛一冰醋酸溶液和高氯酸最适用量分别从0．2mL和0．5mL优化为0．1mL和0．8mL，显色稳定范围确定为10~30min。运用优化的检测方法可以获得很好的标准曲线，提高了R2值，且重复性好、回收率高，可作为灵芝孢子粉总三萜含量检测的一个简便、快速、准确的检测技术。Selected parameters adopted as part of the Chinese Ministry of Agriculture＇s NY/SJ 339-2001 standard protocol, the most common method used for quantifying total triterpenoids in Ganoderrna lucidum, have been optimized. Experimental error derived from operating with small sample volumes was decreased by adopting larger sample volumes （0.1 - 0.6 mL compared with 0.02 - 0.1 mL used in the original protocol） representing the same range of sample concentration. A standard oleanolic acid solution of 0. 2 mg/mL （instead of 1.0 mg/mL used in the original protocol） was adopted for the same purpose. A volatilization step to remove the methanol solvent used to dissolve the oleanolic acid reagent has also been introduced to avoid residual solvent affecting the final concentration of the reactants in the assay mixture. Increased sensitivity, and an absorbance value at 550 nm that was stable for between 10 and 30 min after initiating the reaction, was achieved using 0.1 mL 5% vanillin-glacial acetic acid and 0.8 mL 72% perchloric acid solution. The optimized protocol resulted in an improved standard curve （R2 value increased from 0. 9951 to 0. 9993）, higher recovery levels, greater reproducibility, and provided a convenient, rapid and accurate method for quantifying total triterpenoids in G. lucidum.

关 键 词：灵芝 总三萜含量 优化 

分 类 号：TS207.3[轻工技术与工程—食品科学] R286.0[轻工技术与工程—食品科学与工程]