Protein binding assays for an accurate differentiation of vitamin B12 from its inactive analogue. A study on edible cricket powder.
Protein binding assays for an accurate differentiation of vitamin B12 from its inactive analogue. A study on edible cricket powder.
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蛋白质结合测定法，以准确分化维生素B12与其无活性类似物的分化。一项关于食用板球粉的研究。
DOI：
10.1016/j.fochx.2023.100824
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发表时间：
2023-10-30
期刊：
影响因子：
--
通讯作者：
Mason JB
中科院分区：
文献类型：
--
作者：
Fedosov SN;Nexo E;Heegaard CW;Goldin J;Mason JB
微生物检测高估了昆虫食品中的维生素B12。一种带有蛋白质内在因素的简便方法可以准确地量化B12。使用新开发的校正系数进一步提高了特异度。在可食用的蟋蟀粉中发现低水平的维生素B12：0.75-2.2gμg/100g。相比之下，存在过多的非功能性B12类似物：40-60g/100g。如果使用传统的微生物测量，非活性的维生素B12类似物(钴胺，Cbl)可以模拟食物中的活性Cb1。因此，最近利用免疫亲和吸附、高效液相色谱分离和质谱学(https://doi.org/10.1016/j.foodchem.2021.129048).技术在食用昆虫中发现了高估的cbl。在这里，我们展示了一种简便的结合试验来评估可食用板球粉中Cbl的实用性。该检测使用了Cbl特异的蛋白质固有因子(IF)和类似检测蛋白质结合角蛋白。过量的类似物对IF的亲和力较弱，导致Cbl被适度高估。这种过高的估计被一种新的数学程序纠正了，该程序基于样本中模拟/Cbl的比率以及它们对IF的相对亲和力。我们发现，100g板球粉中含有40-60g类似物和0.75-2.2gμ的Cbl。这一结果得到了高效液相色谱的证实。正确的Cbl测量方法对于任何含有类似物的食物的营养评估都是至关重要的。
Microbiological assays overestimate the vitamin B12 in insect food products. A convenient protocol with the protein intrinsic factor accurately quantifies B12. The specificity is further improved by using a newly-developed correction coefficient. A low level of vitamin B12 is found in edible cricket powder: 0.75–2.2 μg/100 g. In contrast, excessive nonfunctional B12-analogues are present: 40–60 µg/100 g. Inactive analogues of vitamin B12 (cobalamin, Cbl) can mimic the active Cbl in food if using the traditional microbiological measurements. Thus, overestimated Cbl was recently revealed in edible insects employing immunoaffinity adsorption, HPLC-separation and mass spectrometry (https://doi.org/10.1016/j.foodchem.2021.129048). Here we demonstrate the utility of a convenient binding assay to evaluate Cbl in edible cricket powders. The assay employed the Cbl-specific protein intrinsic factor (IF) and the analogue-detecting protein haptocorrin. The excessive analogues had a weak affinity for IF, resulting in a modest overestimate of Cbl. This overestimate was corrected by a novel mathematical procedure, based on the ratio of analogue/Cbl in the sample and their relative affinities for IF. We found that 100 g of cricket powders contained 40–60 µg of analogues and 0.75–2.2 μg of Cbl. This result was confirmed by HPLC. A correct approach to Cbl-measurements is essential for nutritional assessment of any analogue-containing food.
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影响因子：
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作者：
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通讯作者：
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