槲寄生与光合细菌转化槲寄生培养液中总三萜类化合物与齐墩果酸的含量测定.doc

槲寄生与光合细菌转化槲寄生培养液中总三萜类化合物与齐墩果酸的含量测定 摘 要 目的：建立测定槲寄生与光合细菌转化槲寄生培养液中总三萜类化合物与齐墩果酸含量的方法，并对各个样品的含量进行比较。方法：采用紫外分光光度法测定总三萜类化合物的含量，以齐墩果酸为对照品，以5％香草醛冰醛酸溶液、高氯酸为显色系统，检测波长为548 nm；采用高效液相色谱法测定齐墩果酸的含量，色谱柱为Diamonsil C18（200 mm×4.6 mm，5 μm），流动相为甲醇-0.18％磷酸水溶液（86∶14），检测波长为210 nm，柱温为25 ℃。结果：测定总三萜类化合物时，齐墩果酸检测浓度的线性范围为2～12 μg·mL-1（r＝0.995 3）；测定齐墩果酸时，齐墩果酸检测浓度的线性范围为0.41～4.1 μg·mL-1（r＝0. 999 1）。槲寄生75％的乙醇提取物通过球形红细菌、沼泽红假单胞菌分别转化后，总三萜类化合物的含量分别提高36.0％、14.7％，齐墩果酸的含量分别提高880.0％、260.0％。结论：2种光合细菌对槲寄生醇提物的转化均可以提高总三萜和齐墩果酸的含量，推测可能诱导了相关酶的生成。本试验可为光合细菌转化槲寄生化学成分及其转化机制的研究奠定基础。 关键词 槲寄生；光合细菌；三萜类化合物；齐墩果酸；含量 Content Determination of Total Triterpenes Compound and Oleanolic Acid in Viscum coloratum and Its Photosynthetic Bacterium Transformation Products ABSTRACT OBJECTIVE：To establish the methods for the content determination of total triterpenes compound and oleanolic ac- id in Viscum coloratum and its photosynthetic bacterium transformation products，and to compare the contents of each sample. METHODS：With oleanolic acid as control，total triterpenes compound was measured by UV spectrophotometry with 5％ vanil- lin-glacial acetic acid solution and perchloric acid as color-developing agent at detection wavelength of 548 nm. The content of olea- nolic acid was determined by HPLC. The determination was performed on Diamonsil C 18 （200 mm×4.6 mm，5 μm）column with mobile phase consisted of methanol-0.18％ phosphoric acid water（86 ∶ 14）with the detection wavelength at 210 nm and the column temperature at 25 ℃. RESULTS：The linear range of total triterpenes compound was 2～12 μg · mL -1 （r＝0.995 3），and the linear range of oleanolic acid was 0.41～4.1 μg · mL -1 （r＝0.999 1）. 75％ ethanol extracts of V. coloratum was transformed by Rhodobacter sphaeroides and Rhodopseudomonas palustris respectively. The contents of the total triterpenes increased by 36.0％ and 14.7％，and the contents of oleanolic acid increased by 880.0％ and 260.0％，respectively. CONCLUSION：The contents of total triterpenes and oleanolic acid in 75％ ethanol extracts of