恩拉霉素生物检测方法的改进

Improved Biotin Detection Method of Enduracidinmold

试验对国家标准中恩拉霉素的生物检测方法进行了改进,以干燥滤纸片法代替管碟法对不同样品中的恩拉霉素含量进行了测定;并研究了浸提液组成、浸提时间、检定菌浓度等因素对检测结果的影响,建立了恩拉霉素浓度与抑菌圈直径之间的线性方程。研究结果表明,利用丙酮浸提液A(丙酮:1mol/mL HCl∶水=35∶12∶56,用1mol/mL HCl将pH调至3.0)浸提供试品,浸提时间3.5h,可以充分浸提供试品中的恩拉霉素;使用生物测定指示菌CMCC(B)63501芽孢数为3.4×107个/mL的平皿,形成的抑菌圈边缘规则清晰,准确度好;使用干燥滤纸片法可有效消除浸提液或酸性稀释液本身对检测菌株的抑制作用,且操作简单、重复性好,可以同时对大批量的样品进行恩拉霉素含量的测定。恩拉霉素标准溶液在浓度为150~500U/mL的范围内,其产生的抑菌圈直径与效价有较好的线性回归性,所得回归方程:Y=0.00944X+11.55344,相关系数R=0.9962,最终有效区间是0.75~2.5U,测得的结果更接近真实值,可作为一种高效、快速的恩拉霉素含量测定方法,值得推广和使用。

The national standard pinella mildew element biological detection method was improved,using dry filter paper method instead of cylinder plate method,the potassium content of different samples of pinella mold were determined,and the extract composition,extraction time,the concentration of bacteria test and other factors influence on the test results of the established enramycin between concentration and the antibacterial circle diameter of linear equations was studied.The results showed that using acetone extraction liquid acetone（A∶1mol/mL HCl∶water=35∶12∶56,pH value was adjusted to 3.0with 1mol/mL HCl）leaching provided sample,extraction time of 3.5hcould fully immersed test of enramycin.Use of biological indicator bacteria CMCC（B）63501spore count of 3.4×10^7 per mL of the measurement of living flat dish,the formation of the inhibitory circle edge rule was clear,good accuracy.Using dry filter paper method could effectively eliminate the extraction or dilute acidic fluid itself to detect strains inhibitory effect,and the utility model had the advantages of simple operation,good repeatability.It could be used for large quantities of samples of enramycin content determination.Enramycin standard solution at concentrations for 150-500 U/mL and its bacteriostatic circle diameter and valence have good linear regression.The resulting regression equation was Y=0.00944X＋11.55344,the correlation coefficient was R=0.9962,the final effective range was 0.75-2.5U,the measured results were more close to the true value.It could be used as an efficient enduracidin determination method,worthy of promotion and use.