Abstract

Chitin, a biopolymer present in fungi and arthropods, is a compound of interest for various applications, such as in the agricultural and medical fields. With the recently growing interest in the development of insect farming, the availability of chitin-containing residual streams, particularly the molting skins (exuviae), is expected to increase in the near future. For application purposes, accurate quantification of chitin in these insect sources is essential. Previous studies on chitin extraction and quantification often overlooked the purity of the extracted chitin, making the outcomes inconsistent and prone to overestimation. The present study aims to determine chitin content in the exuviae of three insect species mass-reared worldwide: black soldier fly (BSF), mealworm, and house cricket. Chitin was chemically extracted using acid and alkali treatments to remove minerals and proteins. The purity of extracted chitin was evaluated by hydrolyzing the chitin into glucosamine, followed by quantitative determination of the latter using two liquid chromatography methods: electrochemical detection (ECD) and tandem mass spectrometry (MS/MS). Both methods proved accurate and precise, without the need for labor-intensive derivatization steps. Pearson's correlation and Bland-Altman plots showed that the glucosamine determination results obtained by the two methods were comparable, and there is no consistent bias of one approach vs. the other. The chitin content in extracted residues ranged between 7.9 and 18.5%, with the highest amount found in BSF puparium. In summary, the study demonstrated that (1) the residual streams of the insect farming industry have a great potential for utilization as an alternative chitin source, and (2) both LC-ECD and LC-MS/MS are reliable for the quantitative determination of glucosamine in insect chitin.

Highlights

  • Chitin is a naturally occurring polysaccharide composed of N-acetyl-D-glucosamine units linked by β-(1-4) glycosidic bonds and is the second most abundant polymer on Earth after cellulose (Roberts, 1992)

  • The present study aims to compare the electrochemical detection (ECD) with an established mass spectrometry (MS)/MS method and discuss the applicability of the methods to determine chitin content in insect sources

  • The purity of extracted chitin was evaluated by hydrolyzing the chitin into its constituents, followed by quantifying the latter with liquid chromatography methods

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Summary

Introduction

Chitin is a naturally occurring polysaccharide composed of N-acetyl-D-glucosamine units linked by β-(1-4) glycosidic bonds and is the second most abundant polymer on Earth after cellulose (Roberts, 1992). Insect-based bioconversion offers practical solutions to waste management while at the same time presenting an opportunity to produce protein in a sustainable manner (Dossey et al, 2016; van Huis, 2021) This sector has recently witnessed rapid growth worldwide, with an expected volume of 730.000 metric tons by 2030 (Anon, 2019). The residual streams of the insect farming industry, namely the excrements (frass) and the molting skins (exuviae), are expected to become more readily available The latter is most interesting as a source for chitin purification because exuviae contain a considerable amount of chitin, especially in the innermost layer (procuticle) (Kramer et al, 1995; Roer et al, 2015)

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