鲎试剂法测定毕格犬血清中香菇多糖的含量

Quantitative detection of lentinan in Beagle dog serum with limulus amebocyte lysate

目的:对鲎试剂显色基质法测定毕格犬血清中香菇多糖含量的方法进行探讨。方法:香菇多糖、鲎试剂、鲎三肽在一定的条件下反应释放出对硝基苯胺(PNA),溶液呈现黄色,再加入显色剂,呈现玫瑰红色,于550nm波长处比色测定吸光度,其与香菇多糖浓度呈线性关系,从而定量测定出香菇多糖的含量。结果:香菇多糖浓度在5～200ng.mL-1范围内线性关系良好,相关系数0.9922;最低定量检测限为5ng.mL-1;低、中、高3种浓度的日内精密度分别为15.4%,9.8%,4.6%;日间精密度分别为14.3%,9.3%,5.9%;相对回收率为(99.1±9.8)%;绝对回收率为(99.0±4.7)%;血清样品在4℃和-20℃放置条件下7d内稳定性良好。结论:鲎试剂显色基质法具有灵敏度高,准确、快速,重复性好,经济实用等特点,可用于毕格犬血清中香菇多糖含量的测定。

Objective：To study the chromogenic substrate limulus amebocyte lysate（LAL） to detect the content of lentinan in Beagle dog serum.Methods：Lentinan,LAL and the middle three limulus peptide substrate interacted to release paranitroaniline（PNA） under a proper condition.PNA with some reagents will produce a rose red color.The solution was measured photometrically at 550 nm.The amount of PNA released is in proportion to the amount of lentinan,so the method can be used in quantitative detection of lentinan in the samples.Results：The range of linearity was 5-200 ng·mL-1;The correlation coefficient of standard curve was greater or equal to 0.9900;The RSDs of intra-day precision tests were 15.4%,9.8%,4.6%;The RSDs of inter-day precision tests were 14.3%,9.3%,5.9%;The average relative recovery was （99.1±9.8）%;The average absolute recovery was （99.0±4.7）%;The stability was very good.Conclusion：LAL is proved to be sensitive,accurate,quick,repeatable,economical and practical in detection of lentinan in the serum of Beagle dogs.