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药用及食用蝗虫中多种霉菌毒素的污染部位及残留水平

Contamination Parts and Residue Levels of Multi-Mycotoxins in Medicinal and Edible Locust.

作者信息

Kong Dandan, Kong Weijun, Yang Xiaoli, Yang Meihua

机构信息

Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences - Peking Union Medical College, Beijing, China.

出版信息

Front Pharmacol. 2018 May 17;9:480. doi: 10.3389/fphar.2018.00480. eCollection 2018.

Abstract

Locust is esteemed as a traditional Chinese medicine, as well as one of the most important nutritional foods especially in Asian countries. However, some toxic secondary metabolites such as mycotoxins are usually found in different parts of locust to affect its quality and safety. This study aimed to investigate the aflatoxins (AFs) contaminated parts by observing , spores' diameter, amount and distribution on head, tentacle, wing, belly and shank parts of the locust with scanning electron microscopy (SEM). Furthermore, to assess the residue levels of multi-mycotoxins in the locust, the high performance liquid chromatography with fluorescence detection (HPLC-FLD) was adopted. The technique was used to determine the contents of AFs, zearalenone (ZON) and α-zearalenol (α-ZOL) in locust and the positive samples were confirmed by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The chromatographic conditions, MS/MS parameters and the method of sample extraction were carefully optimized. Results revealed that obvious differences of strains and spores were found, while the spores' diameter ranged from 3.0 to 13.0 μm in different contaminated parts of the locust samples. The HPLC-FLD method for multi-mycotoxins analysis showed good selectivity, linearity, recovery and precision. Limits of quantification (LOQs) were lower than 27.6 μg/kg, while limits of detection (LODs) were in the range of 0.02-8.6 μg/kg. The accuracy of the developed method was validated regarding recoveries of 80.1-118.1% with relative standard deviation (RSD) ≤ 11.4%. Finally, the developed multi-mycotoxin method was applied for screening of these mycotoxins in 11 commercial locust samples. Only AFB and AFB were found in six samples, and the contamination levels ranged from 0.12 to 4.4 μg/kg, which were lower than the maximum residue limit and can be used safely. This is the first report on the exploration of contamination parts and levels of multi-mycotoxins in medicinal and edible locust. The combined method of SEM and HPLC-FLD exhibited advantages of low cost, high sensitivity, rapid determination, convenience and especially intuitive judgment, which is proposed for contamination parts observation, for the large-scale quantification of multi-mycotoxins in other medicinal animal matrices.

摘要

蝗虫被视为一种传统中药,也是最重要的营养食品之一,尤其在亚洲国家。然而,蝗虫的不同部位通常会发现一些有毒的次生代谢产物,如霉菌毒素,这会影响其质量和安全性。本研究旨在通过扫描电子显微镜(SEM)观察蝗虫头部、触角、翅膀、腹部和腿部的霉菌孢子直径、数量及分布,以调查黄曲霉毒素(AFs)的污染部位。此外,为评估蝗虫中多种霉菌毒素的残留水平,采用了带荧光检测的高效液相色谱法(HPLC-FLD)。该技术用于测定蝗虫中AFs、玉米赤霉烯酮(ZON)和α-玉米赤霉醇(α-ZOL)的含量,阳性样品通过高效液相色谱-串联质谱法(HPLC-MS/MS)进行确证。对色谱条件、MS/MS参数和样品提取方法进行了仔细优化。结果表明,在蝗虫样品的不同污染部位发现了明显不同的菌株和孢子,孢子直径在3.0至13.0μm之间。用于多种霉菌毒素分析的HPLC-FLD方法具有良好的选择性、线性、回收率和精密度。定量限(LOQs)低于27.6μg/kg,而检测限(LODs)在0.02-8.6μg/kg范围内。所建立方法的准确性通过回收率为80.1-118.1%、相对标准偏差(RSD)≤11.4%得到验证。最后,将所建立的多种霉菌毒素方法应用于11个市售蝗虫样品中这些霉菌毒素的筛查。仅在6个样品中发现了AFB和AFB,污染水平在0.12至4.4μg/kg之间,低于最大残留限量,可以安全使用。这是关于药用和食用蝗虫中多种霉菌毒素污染部位和水平探索的首次报告。SEM和HPLC-FLD的联合方法具有成本低、灵敏度高、测定快速、方便,尤其是判断直观等优点,建议用于污染部位观察以及其他药用动物基质中多种霉菌毒素的大规模定量分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9e4/5966561/cd455ea48567/fphar-09-00480-g001.jpg

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