Developing an in-house colorimetric method for detecting organophosphate and carbamate residue using cricket cholinesterase

IF 5.2 2区农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY

Chemical and Biological Technologies in Agriculture Pub Date : 2023-07-27 DOI:10.1186/s40538-023-00442-3

Anurak Wongta, Surat Hongsibsong, Priyanshi Anand, Udomsurb Jaitum, Nootchakarn Sawarng, Wasin Wongwilai, Pongsathorn Dhumtanom

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引用次数: 0

Abstract

Background

Pesticides such as organophosphates and carbamates are used to control pests, leaving residues on fruits and vegetables. The assessment of residues in food materials is required for consumer and agricultural worker health protection. The purpose of this study was to develop an in-house colorimetric method for detecting organophosphate and carbamate residues using cricket cholinesterase that can be used as a low-cost, simple-to-use, and widely used test.

Results

The method’s parameters, including substrate specificity, kinetics, and incubation time, were optimized. The results indicated that acetylthiocholine iodide was more active than butyrylthiocholine iodide. Using a buffer with a pH of 8.0 resulted in the greatest activity after 5 min. The same enzyme activity was observed at 25 °C and 30 °C, which was greater than at 35 °C and 40 °C. In this study, the Km and Vmax values for cricket cholinesterase were 3.75 mM and 35 µM/min, respectively. The specificity and purification factor of the produced ChE were 1.54 µM/min/mg and 1.43-fold, respectively. The limit of detection for detecting organophosphates and carbamates was in the range of 0.002–0.877 ppm. The validity of the method yielded the following results: relative sensitivity of 95%, relative specificity of 90%, relative accuracy of 93%, positive predictive value of 95%, and negative predictive value of 90%.

Conclusions

The developed method is simple and inexpensive, and it can analyze organophosphates and carbamates in a variety of vegetables at an appropriate detection level. Cholinesterase from cricket has the potential to be used for the development of a variety of methods for detecting residues, and the sensitivity could be enhanced using new techniques in the future.

Graphical Abstract

查看原文本刊更多论文

利用蟋蟀胆碱酯酶建立了一种检测有机磷和氨基甲酸酯残留的室内比色法

有机磷和氨基甲酸酯等杀虫剂被用来控制害虫，在水果和蔬菜上留下残留物。为了保护消费者和农业工人的健康，需要对食品材料中的残留物进行评估。本研究的目的是开发一种利用蟋蟀胆碱酯酶检测有机磷酸酯和氨基甲酸酯残留的内部比色法，该方法可作为一种低成本，简单易用且广泛使用的检测方法。结果优化了该方法的底物特异性、动力学和孵育时间等参数。结果表明，乙酰硫代胆碱的活性高于丁基硫代胆碱。使用pH为8.0的缓冲液，5 min后酶活性最高。25°C和30°C条件下酶活性相同，大于35°C和40°C条件下。本研究蟋蟀胆碱酯酶的Km和Vmax分别为3.75 mM和35µM/min。所得ChE的特异性为1.54µM/min/mg，纯化系数为1.43倍。有机磷酸盐和氨基甲酸酯的检出限在0.002 ~ 0.877 ppm范围内。方法的效度为:相对灵敏度95%，相对特异性90%，相对准确度93%，阳性预测值95%，阴性预测值90%。结论该方法操作简便，成本低廉，可在适当的检测水平下对多种蔬菜中的有机磷和氨基甲酸酯类进行检测。蟋蟀胆碱酯酶具有开发多种残留检测方法的潜力，未来可通过新技术提高灵敏度。图形抽象

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来源期刊

Chemical and Biological Technologies in Agriculture Biochemistry, Genetics and Molecular Biology-Biotechnology

CiteScore

6.80

自引率

3.00%

发文量

审稿时长

15 weeks

期刊介绍： Chemical and Biological Technologies in Agriculture is an international, interdisciplinary, peer-reviewed forum for the advancement and application to all fields of agriculture of modern chemical, biochemical and molecular technologies. The scope of this journal includes chemical and biochemical processes aimed to increase sustainable agricultural and food production, the evaluation of quality and origin of raw primary products and their transformation into foods and chemicals, as well as environmental monitoring and remediation. Of special interest are the effects of chemical and biochemical technologies, also at the nano and supramolecular scale, on the relationships between soil, plants, microorganisms and their environment, with the help of modern bioinformatics. Another special focus is the use of modern bioorganic and biological chemistry to develop new technologies for plant nutrition and bio-stimulation, advancement of biorefineries from biomasses, safe and traceable food products, carbon storage in soil and plants and restoration of contaminated soils to agriculture. This journal presents the first opportunity to bring together researchers from a wide number of disciplines within the agricultural chemical and biological sciences, from both industry and academia. The principle aim of Chemical and Biological Technologies in Agriculture is to allow the exchange of the most advanced chemical and biochemical knowledge to develop technologies which address one of the most pressing challenges of our times - sustaining a growing world population. Chemical and Biological Technologies in Agriculture publishes original research articles, short letters and invited reviews. Articles from scientists in industry, academia as well as private research institutes, non-governmental and environmental organizations are encouraged.