Nakata Ryu, Yoshinaga Naoko, Teraishi Masayoshi, Okumoto Yutaka, Mori Naoki
a Graduate School of Agriculture, Kyoto University , Kyoto , Japan.
Biosci Biotechnol Biochem. 2019 Sep;83(9):1624-1629. doi: 10.1080/09168451.2019.1611407. Epub 2019 May 3.
Chitin, poly (β-(1→4)--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-I 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 (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.
几丁质，即聚（β-(1→4)-N-乙酰-D-葡萄糖胺），是昆虫体内一种重要的生物聚合物，用作围食膜的主要成分。尽管几丁质难以定量，但从害虫防治的角度来看，围食膜中的几丁质含量值得关注。在本研究中，我们建立了一种通过图像处理定量围食膜中几丁质的简便方法。在该方法中，几丁质通过壳聚糖-I复合物间接定量，该复合物呈现出特定的红紫色。使用壳聚糖溶液绘制的校准曲线在0.05-0.5μg/μL的浓度范围内显示出良好的线性。我们使用该方法对夜蛾科幼虫围食膜中的几丁质含量进行了定量。在整个研究过程中，仅使用了常见的廉价试剂和易于获得的仪器。该方法可被广泛的研究人员轻松应用于材料中几丁质和壳聚糖含量的灵敏定量。缩写：LOD：检测限；LOQ：定量限；ROI：感兴趣区域；RSD：相对标准偏差。
