Lc 20 ad t

Proximate analysis was done following official methods of analysis of AOAC International⁸ . The moisture and dry matter contents were determined using a hot air oven by drying the sample at 105 °C for 8 hours. Ash content was determined at 600 °C. Nitrogen content was estimated by Kjeldahl method (Kelplus Classic, DX VA, Pelican Equipment) and crude protein was calculated by multiplying nitrogen percentage by 6.25. Lipid was determined by the solvent extraction method by Soxtec system (SOCS Plus, SCS-6, Pelican Equipment) using diethyl ether (boiling point, 40–60 °C) as a solvent. The crude fiber was determined by digesting the moisture and fat-free sample successively with dilute (1.25%) acid and alkali using Fibre cap (Foss tecator, Sweden). Nitrogen free extract (NFE) was determined as per the following formula $$\mathrm{NFE}(\%)=100–(\%\mathrm{moisture}+\%\mathrm{crude}\mathrm{protein}+\%\mathrm{crude}\mathrm{lipid}+\%\mathrm{crude}\mathrm{fiber}+\%\mathrm{ash})$$
Fatty acid, amino acid, and mineral analysis were carried out following AOAC⁸ . Calcium, magnesium, chromium, manganese, iron, chromium analysis was performed in Absorption Spectrophotometer (Model 280 FS- Make Agilent Technology). Phosphorus content was estimated using UV-Vis Spectrophotometer (Model - Agilent Carry-60). Amino acids were analyzed using HPLC (Model - Shimadzu LC-20 AD/T)). Fatty acids were analyzed through gas chromatography (Model - Shimadzu 2010 Plus).