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The mixture was subjected to sonication (5 min) and centrifugation (3 min, 1800 rpm).
The hybrid mixture was subjected to ultrasonication for effective blending.
These were excised and the protein mixture was subjected to tryptic digestion and LC-MS/MS analysis.
This mixture was subjected to semi-preparative HPLC purification.
After incubation, 10 μl of the reaction mixture was subjected to 20 % SDS-PAGE.
This mixture was subjected to column chromatography for the separation of individual acids.
The obtained mixture was subjected to SAFE distillation [22] and extracted with dichloromethane (2 × 200 mL).
Initially, the mixture was subjected to dynamic vacuum for 1 h.
Subsequently, the mixture was subjected to the Soxhlet extraction technique to obtain the extractive compound.
The mixture was subjected to autoclaving at 121 °C and 15 psi for 15 min.
The mixture was subjected to stirring at room temperature for 2 h.
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