An easy, inexpensive, and sensitive method for the quantification of chitin in insect peritrophic membrane by image processing

Author:

Nakata Ryu1,Yoshinaga Naoko1,Teraishi Masayoshi1,Okumoto Yutaka1,Mori Naoki1

Affiliation:

1. Graduate School of Agriculture, Kyoto University, Kyoto, Japan

Abstract

ABSTRACT Chitin, poly (β-(1→4)-N-acetyl-d-glucosamine), is an important biopolymer for insects that is utilized as a major component of peritrophic membrane. The chitin content in peritrophic membrane is of expedient interest from a pest control perspective, although it is hard to quantify chitin. In this study, we establish a facile method for the quantification of chitin in peritrophic membrane by image processing. In this method, chitin was indirectly quantified using chitosan–I3− complex, which exhibited a specific red-purple color. A calibration curve using a chitosan solution showed good linearity in a concentration range of 0.05–0.5 μg/μL. We quantified the amount of chitin in peritrophic membrane of Spodoptera litura (Lepidoptera: Noctuidae) larvae using this method. Throughout the study, only common inexpensive regents and easily attainable apparatuses were employed. This method can be easily applied to the sensitive quantification of the amounts of chitin and chitosan in materials by wide range of researchers. Abbreviations: LOD: limit of detection; LOQ: limit of quantification; ROI: region of interest; RSD: relative standard deviation.

Funder

Grants-in-Aids for Scientific Research

JSPS Research Fellowship for Young Scientist

JSPS KAKENHI

Publisher

Oxford University Press (OUP)

Subject

Organic Chemistry,Molecular Biology,Applied Microbiology and Biotechnology,General Medicine,Biochemistry,Analytical Chemistry,Biotechnology

Reference35 articles.

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