以高效液相層析／比色分析組合法定量Penicillium sclerotiorum生產之Sclerotiorin   全文下載

HPLC/Colorimetry Combination Method for Quantitative Analyses of Sclerotiorin Produced by Penicillium sclerotiorum

本研究擬建立一種在無法獲得純標準品之情況下，利用HPLC配合比色法（HPLC／colorimetry combination method）定量Penicillium sclerotiorum菌體中sclerotiorin含量之方法。菌體以甲醇萃取後，採用C18逆相管柱進行HPLC分析（流動相為乙?／水 = 65／35，v／v；檢測波長為370 nm）同時進行比色分析（測定OD370）， 可以HPLC測得之面積百分比和比色法測得之OD370之關係所導出之方程式計算出菌體中sclerotiorin之含量。結果顯示，最低檢測濃度（LOQ）為0.5μg／mL，回收試驗之回收率為97-109%，變異係數均在3.8%以下，分析一個樣品，僅需約30min，相當簡便。

In this study, quantitative analysis of sclerotiorin from Penicillium sclerotiorum was established by using HPLC/colorimetry combination method at the condition of unavailability of authentic sclerotiorin compound. Mycelium was extracted with methanol, then analyzed by HPLC (a C18 reverse phase column with acetonitrile/water = 65/35, v/v as mobile phase and detected at 370 nm) and simultaneously measured OD370 with spectrophotometers. The limit of quantitation (LOQ) was 0.5 μg/mL. Recovery studies were performed using the lyophilized mycelia, with each spiked with sclerotiorin at 1 and 10 mg, respectively. The recoveries were found in the range of 97~109%. The coefficients of variation were less than 3.8% with this HPLC / OD370 combination method. 本研究擬建立一種在無法獲得純標準品之情況下，利用HPLC配合比色法（HPLC／colorimetry combination method）定量Penicillium sclerotiorum菌體中sclerotiorin含量之方法。菌體以甲醇萃取後，採用C18逆相管柱進行HPLC分析（流動相為乙腈／水 = 65／35，v／v；檢測波長為370 nm）同時進行比色分析（測定OD370）， 可以HPLC測得之面積百分比和比色法測得之OD370之關係所導出之方程式計算出菌體中sclerotiorin之含量。結果顯示，最低檢測濃度（LOQ）為0.5μg／mL，回收試驗之回收率為97-109%，變異係數均在3.8%以下，分析一個樣品，僅需約30min，相當簡便。