河鲀鱼基质中河豚毒素标准样品的研制

Preparation of matrix reference material for tetrodotoxin in Tetraodontidae

中文关键词:  河鲀鱼基质  河豚毒素  基体标准样品  制备

英文关键词:Tetraodontidae matrix  tetrodotoxin  matrix reference material  preparation

中文摘要:

      目的 研制基于河鲀鱼基质的河豚毒素(tetrodotoxin, TTX)定量分析标准样品。方法 以野生河鲀鱼肝脏、卵巢、精巢和肌肉组织为基料, 利用正己烷脱脂分离处理后, 经两次冷冻干燥制备河鲀鱼基质的TTX标准样品, 采用液相色谱-串联质谱法进行定性确证。样品分装成500瓶后, 进行均匀性、稳定性检验和定值分析。结果 经确证该基体标准样品中含有TTX; 从基体标准样品中随机抽取15瓶进行均匀性检验, 经F检验表明在95%的置信区间范围内样品均匀性良好; 长期稳定性考察在0~4 ℃低温条件下历经了长时间保存, 短期稳定性在-80~55 ℃范围内进行运输条件下的考察, 结果表明在考察期间内样品稳定性良好; 经国内8家实验室进行协同定值, 评定了定值结果的不确定度, 该基体标准样品中TTX特性值及不确定度为(5.20±0.03) mg/kg (k=2)。 结论 该基体标准样品达到国家标准样品的技术要求, 可用于对水产品中河豚毒素检测过程的方法验证与质量控制等活动。

英文摘要:

      Objective To develop a matrix reference material for quantitative analysis of tetrodotoxin (TTX) based on Tetraodontidae matrix. Methods Based on puffer fish matrix, the liver, ovary, testis and muscle tissue of wild puffer fish were used as base materials, degreased and separated with n-hexane, and then freeze-dried twice to prepare the matrix reference material of TTX in Tetraodontidae, which was qualitatively confirmed by liquid chromatography-tandem mass spectrometry. After the sample was sub packed into 500 bottles, the uniformity, stability test and fixed value analysis were carried out. Results The matrix reference materia had been confirmed to contain TTX; 15 bottles were randomly selected from the matrix reference material for uniformity test, and the F test showed that the sample uniformity was good within the confidence interval of 95%; the long-term stability was investigated under the low temperature of 0-4 ℃ for a long time and the short-term stability was investigated under the transportation condition within the range of -80-55 ℃, the results showed that the stability of the samples were good during the investigation; the collaborative setting by 8 laboratories in China were conducted to assess the uncertainty of the calibration result, the TTX characteristic value and uncertainty of the matrix reference material were (5.20±0.03) mg/kg (k=2). Conclusion The matrix reference material meets the technical requirements of the national reference material, and can be used for method verification and quality control of TTX detection process in aquatic products.