纳他霉素效价测定方法的改良及含量的快速测定

An Improved Method for Natamycin Titer Measurement and the Rapid Determination of Its Content

摘要:

为了确定固体发酵过程中纳他霉素含量快速测定和生物效价准确测定方法，本文采用高效液相色谱法、紫外分光光度法和管碟法测定了同一批褐黄孢链霉菌固体发酵产物纳他霉素的含量。其中高效液相色谱法和紫外分光光度法测定纳他霉素固体发酵产物含量分别为2351.52 mg/L和1979.87 mg/L，前者结果准确而稳定。后者测定准确率为96.97%（以高效液相色谱的测定结果为100%计）。适用于固体发酵过程中的纳他霉素含量的快速测定。管碟法测定纳他霉素平均效价为1979.87 mg/L。改良管碟法采用正己烷封盖纳他霉素溶剂甲醇后，测定的平均效价达到1990.29 mg/L，准确率由84.20%提高到84.64%（以高效液相色谱的测定结果为100%计）。采用正己烷封盖能有效控制纳他霉素溶剂甲醇的挥发性并提高管碟法的准确性和测定的浓度范围，而正己烷本身不具有抑菌效果。

Abstract:

To find an effective method for the rapid detection of natamycin and an accurate analysis of biological potency during solid-phase fermentation, three methods including high-performance liquid chromatography (HPLC), ultraviolet spectroscopy, and the cylinder-plate method were used to determine the natamycin content in the product of a batch of Streptomyces glvosporue during solid-state fermentation. The natamycin content in the solid fermentation product determined by HPLC and ultraviolet spectroscopy was 2351.52 mg/L and 2280.16 mg/L, respectively. The former showed good accuracy and reproducibility, while the accuracy of latter was 96.97% of the former (assigning the accuracy of HPLC method as 100%) and proved to be applicable for the rapid determination of natamycin content during solid fermentation. The average potency of natamycin determined by the cylinder-plate method was 1979.87 mg/L, and using an improved cylinder-plate method was 1990.29 mg/L when n-hexane was used in combination with methanol as the solvent. The accuracy of HPLC determined result increased from 84.20% to 84.64% using the cylinder-plate method (assigning the accuracy of HPLC method as 100%). The results indicated that n-hexane could effectively control the volatility of methanol, and improved the accuracy and concentration range for measurements using the cylinder-plate method, without having an antimicrobial effect.