An Improved Analytical Method For Determination Of Biogenic Amines In Bee Products And Study Of Their Changes During Processing And Storage

In recent years, with bee products (BPs) more and more popular among customers, the risk assessment draws attention from supervision department. As potential quality and safety risk factors, biogenic amines (BAs) gradually receive great attention and deep investigation. BPs contains high amounts of free amino acids, and be prone to produce BAs when microorganism infected. Excessive intake of BAs may cause detriment to human health. Therefore, there is an urgent need to establish an analytical method for the detection of BAs in BPs, and to provide a technical support for the risk assessment. So far, neither the reports of effective analytical methods for determination of BAs in BPs, nor the survey on change rule of BAs in BPs during processing or storage is available. The aim of this paper is to establish a quick and effective method to determine BAs in honey, royal jelly and bee pollen, and to investigate the effect of hot processing on the stability of BAs in BPs, as well as the change rule of BAs during the storage, and therefore to provide references for the setting of the standard which is for quality evaluation and risk assessment of BPs.This paper contains three sections. A UHPLC-MS/MS method for the determination of 12 common BAs in BPs was established in the first section. The developed method had a good linearity with all the correlation coefficients greater than 0.99 among a certain concentration range. The limits of detection (LODs) and limits of quantification (LOQs) of the method were from 0.4 to 144.4 ?g/kg and from 1.3 to 476.5 ?g/kg, respectively. The recoveries of the samples spiked at 0.1-1.0 mg/kg were in the range of 72 to 116% with RSD of intra-day and inter-day lower than 10% and 15%, respectively. The matrix effect of suppression was observed for the determination of BAs, which could be minimized to an acceptable range by 20 times dilution using pure acetonitrile. This developed method is simple, sensitive and fast. Meanwhile, the method has reliable abilities of qualitification and quantification, and thus can be used for the qualitative and quantitative analysis of BAs in BPs.The varieties and amounts of BAs in the common marketing honey, royal jelly and bee pollen were determined in the second section.8 BAs were detected in honey samples. The total amounts of BAs in honeys were in big difference with a range of 45.4 to 4349.9 ?g/kg. Sample with the highest total amount of BAs was linden honey, while sample with the lowest one was schisandra chinensis honey. Results showed that phenylethylamine (PHE) and tryptamine (TR) were representative BAs of linden honey and jujube honey, respectively, and could be used as potential markers of these two honeys for characterization of botanical origins.9 BAs were detected in royal jelly, with total amounts ranging between 5138.9 and 9228.8 ?g/kg. Total amount of BAs in lyophilized royal jelly powder was higher than that in fresh royal jelly, especially distinct for putrescine (PUT) and dopamine (DOP).8 BAs were detected in bee pollen, with total amount ranging from 6837.4 to 25444.2 ?g/kg. Spermine (SP) and spermidine (SPD) were the two highest BAs in bee pollen. Results showed that a certain content of BAs existed in the common BPs, but they won't be risky for customers based on the acceptable daily intake of BAs, average daily consumption of BPs, and BA amounts determined in BPs. However, the existence of some BAs means that BPs have been infected by microorganism, whose hazards to human health are not clear so far. Therefore, from the viewpoint of ensuring a health diet to customers, it is still necessary to deeply investigate BAs in BPs, and to provide basic data for the formulation of relevant industry standards.The change rule of BAs in BPs during storage and the effect of processing temperature and fermentation on the varieties and amounts of BAs in BPs were preliminarily investigated in the third section. The varieties of BAs in BPs did not alter during the storage, while the amounts of BAs changed a little. Temperature can cause a certain impact on BA amounts. Amounts of BAs in bee pollen improved significantly when processing temperature increasing. Fermentation was also an influence factor of BA amounts. PUT amount in fermented rape bee pollen was apparently higher than that in unfermented one. The significant difference of PUT content in fermented and unfermented bee pollens could be used as a criteria for judging whether the pollen has been infected by bacteria. Therefore, it is essential to further investigate change rule of BAs in BPs during processing and storage, in order to guarantee the customers' health and safety.