Moisture, crude protein, ash, and crude fiber were analyzed according to the standard methods [29 ], and crude lipid extraction was conducted according to the method of Folch et al. [30 (link)]. Moisture was determined by drying the samples at 105°C until a constant weight was measured. Crude protein (N × 6.25) was examined using the Kjeldahl method. Crude lipid was extracted using a chloroform–methanol solvent (2 : 1 v/v), and the solvent was removed using a vacuum concentrator (EYELA, Tokyo, Japan). Ash was measured through incineration in a muffle furnace (Barnstead Thermolyne, Moorhead, MN, USA) at 550°C for 6 h. Crude fiber was measured with a fiber analyzer (ANKOM Technology, Macedon, NY, USA) after digestion with 1.25% sulfuric acid and 1.25% sodium hydroxide solutions. Nitrogen-free extract (NFE) was calculated as 100% − (moisture + crude protein + crude lipid + ash + crude fiber). Gross energy was calculated as 16.7, 37.4, and 16.7 kJ g−1 for crude protein, crude lipid, and NFE, respectively [31 ]. The amino acid composition was determined using a high-speed amino acid analyzer (HITACHI, Tokyo, Japan) after hydrolyzation in a 4 N methane sulfonic acid at 110°C for 24 h.
High speed amino acid analyzer
Manufactured by Hitachi
Sourced in Japan
The High-speed amino acid analyzer by Hitachi is a laboratory instrument designed for the rapid and accurate analysis of amino acid compositions. It employs a chromatographic technique to separate and quantify individual amino acids within a sample. The core function of this equipment is to provide efficient and reliable amino acid analysis for various applications in research, clinical, and industrial settings.
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2 protocols using high speed amino acid analyzer
1
Proximate and Amino Acid Analysis
2
Chemical Analysis of Fermented Products
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The pH and electrical conductivity (EC) value was measured every week during the whole fermentation period. The pH value was measured with the pH meter (METTLER TOLEDO) and EC value was measured with the EC meter (METTLER TOLEDO). The contents of free amino acids (Aa) in the fermented products were determined by the High Speed Amino Acid Analyzer (Hitachi, Japan) [20 (link)]. The contents of main ions in the fermentation products were determined by the Ion Chromatograph (Thermo Fisher, Germany) [21 (link)]. The content of available N in the fermentation products was determined by Alkaline Hydrolysis Diffusion method [22 (link)]. The content of total phenol was determined by Folin-Ciocalteu colorimetric method [23 (link)] and the content of total acids were determined by Acid-base Titration method.
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