动态显色法鲎试验用于果汁细菌内毒素活性检测及干扰分析

Quantitative detection and influence factors of endotoxin activities in juice products using by Tachypleus amebocyte lysate assay of kinetic chromogenic analysis

目的建立一种定量检测果汁细菌内毒素活性的方法,准确评估果汁潜在的内毒素污染风险。方法采用动态显色法东方鲎(Tachypleus amebocyte lysate, TAL)试验对常见的4种果汁(橙汁、桃汁、苹果汁和葡萄汁)细菌内毒素活性进行检测,对果汁样品中的干扰因素进行识别与排除,最后使用抗增液排除β-葡聚糖等类似物质带来的假阳性结果。结果市售果汁(n=36)的细菌内毒素活性范围为<0.2~209.5 EU/mL,自榨果汁(n=12)的细菌内毒素活性范围为<0.4~4.2EU/mL,加标回收率范围均在50%~200%之间,符合药典要求。结论动态显色法TAL试验适用于常见的4种果汁(橙汁、桃汁、苹果汁和葡萄汁)细菌内毒素活性的检测,为果汁的健康风险评价和卫生监管提供参考。

Objective To establish a quantitative detection method for endotoxin activities in juice, in order to provide technical support for assessment of potential endotoxin contamination risk in juice. Methods The endotoxin activities of juice were detected by Tachypleus amebocyte lysate (TAL) assay of kinetic chromogenic analysis, the interference factors were identified and eliminated, and the juice samples with positive results of TAL assay were verified by endotoxin-specific buffer in order to avoid the false positive caused by β-glucan. Results The endotoxin activities of 4 kinds of juice products (n=36) and self-squeezed juice (n=12) were <0.2~209.5 EU/mL and <0.4~4.2 EU/mL, respectively. The recovery rates met requirements of Chinese Pharmacopoeia (50%~200%). Conclusion The TAL assay of kinetic chromogenic analysis is suitable for the quantitative determination of endotoxin activities in 4 kinds of common fruit juice (orange juice, peach juice, apple juice and grape juice), which provides reference for health risk assessment and health supervision of fruit juice.