BioSystems® Y15 组胺脱氢酶试剂盒用于检测鱼类和渔业产品中组胺的验证研究：AOAC 性能测试方法 SM 072001。

Validation Study of BioSystems® Y15 Histamine Dehydrogenase Kit for the Detection of Histamine in Fish and Fishery Products: AOAC Performance Tested MethodSM 072001.

机构信息

BioSystems S.A., Costa Brava 30, Barcelona, Catalonia, Spain, 08030.

出版信息

J AOAC Int. 2021 Jun 12;104(3):693-711. doi: 10.1093/jaoacint/qsaa139.

DOI:10.1093/jaoacint/qsaa139

PMID:33367653

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8372101/

Abstract

BACKGROUND

BioSystems has developed a histamine kit for automated procedure, through the use of a Y15 analyzer, for quantification of histamine in fish and fishery products.

OBJECTIVE

Validate the method under the specific guidelines of the AOAC Research Institute Performance Tested MethodSM (PTM) program.

METHOD

Samples are extracted with boiling water. The enzymatic method is based on histamine dehydrogenase that catalyzes the oxidation of histamine in the presence of an electron mediator that reduces a dye that is measured at 420 nm. The increase of absorbance is proportional to the histamine concentration. Dispensing of reagents and sample, absorbance readings, calibration, and calculation of results are performed automatically in the analyzer BioSystems Y15.

RESULTS

The linearity ranges from 0 to 200 mg/kg (r2 > 0.99). The LOQ is 10 mg/kg in all the matrixes. Recoveries range from 75 to 107% at concentrations from 5 to 200 mg/kg, with repeatability precision values between 0.8 and 5.5%. Comparisons with the HPLC reference method shows a good correlation. Cross-reactivity of the assay is negligible for all biogenic amines tested except for agmatine (6.3%). Product consistency was verified by validating lot-to-lot variations and variations within the same lot. Shelf life was verified by real-time stability testing during 40 months at 2-8°C. No differences in histamine detection were observed in robustness testing, in which minor changes are introduced to the assay protocol.

CONCLUSIONS

The automated, simple, and rapid BioSystems Y15 Histamine Dehydrogenase Kit has been successfully validated.

HIGHLIGHTS

The method is qualified for PTM certification No. 072001.

摘要

背景

BioSystems 公司开发了一种用于自动化处理的组胺试剂盒，通过使用 Y15 分析仪，可定量检测鱼类和渔业产品中的组胺。

目的

根据 AOAC 研究机构性能测试方法（PTM）计划的具体准则验证该方法。

方法

样品用沸水提取。该酶法基于组胺脱氢酶，在电子介体存在下，该酶催化组胺的氧化，还原染料并在 420nm 处测量吸光度。吸光度的增加与组胺浓度成正比。试剂和样品的分配、吸光度读数、校准和结果计算均在 BioSystems Y15 分析仪中自动进行。

结果

线性范围为 0 至 200mg/kg（r2>0.99）。在所有基质中，LOQ 均为 10mg/kg。在 5 至 200mg/kg 的浓度下，回收率范围为 75%至 107%，重复性精密度值在 0.8%至 5.5%之间。与 HPLC 参考方法的比较显示出良好的相关性。除精胺（6.3%）外，该测定法对所有生物胺的交叉反应性可忽略不计。通过验证批间和同批内的变化，验证了产品一致性。通过在 2-8°C 下实时稳定性测试，验证了保质期为 40 个月。在稳健性测试中，对测定方案进行了微小改动，未观察到组胺检测的差异。