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投稿时间：2015-02-18
投稿时间：2015-02-18
中文摘要: 利用反相高效液相色谱法从 24 种真菌中筛选出对根皮素具有转化作用的菌种并进行底物消耗率的测定。色 谱条件：Welchrom C18 色谱柱（4.6 mm×250 mm，5 μm）；流动相 A 相：冰醋酸 ∶ 水=2 ∶ 98（体积比），B 相∶乙腈 ∶ 水 ∶ 冰醋酸 =80 ∶ 19.6 ∶ 0.4（体积比），梯度洗脱；流速：1.0 mL/min；检测波长：280 nm。根皮素质量浓度在 0.067 1 mg/mL～1.073 6 mg/mL 内线性关系良好，回归方程为 Y= 3.5×107 X + 1.5×106（R2 = 0.999 9）。经 HPLC 检测 4 种真菌对根皮素有转化作用，当液 体培养基中根皮素质量浓度为 0.3 mg/mL 时，测得培养后底物消耗率分别为 60.46 %、43.67 %、91.63 %、40.32 %。该法简 便、准确、重现性好，可适用于根皮素微生物转化研究中非单一转化产物的定性和定量分析。
Abstract:To study 24 species of fungi on the transformation of phloretin and determine the consumption rate of activated fungi by RP-HPLC. The biotransformation reactions of phloretin were identified by RP-HPLC. The optimum HPLC conditions were as follows：column was Welchrom C18（4.6 mm×250 mm， 5 μm）. Gradient elu－ tion with mobile phase A：HAC ∶ H2O =2 ∶ 98（volume ratio） and mobile phase B：MeCN∶H2O∶HAC=80∶19.6∶0.4 （volume ratio） in the gradient program at a flow rate of 1.0 mL/min was utilized after optimization. The detection wavelength was set at 280 nm. The calibration curve was Y= 3.5×107 X + 1.5×106 （R2 = 0.999 9）with the linear in the range of 0.067 1 mg/mL～1.073 6 mg/mL for phloretin. The results showed that phloretin was biotrans－ formed by four kinds of fungi and the consumption rate were 60.46 %， 43.67 %， 91.63 %， 40.32 % when the concentration of phloretin in medium was 0.3 mg/mL. The results indicated that this detective method was stable and accurate for qualitation and quantitation of the microbial conversation products of phloretin.
keywords: phloretin RP-HPLC biotransformation consumption rate
文章编号：201608040 中图分类号： 文献标志码：A
基金项目:国家自然科学基金项目（31201345）；中国博士后基金项目（2014M562674）；天津市自然科学基金项目（13JCYBJC24400）
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