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During the validation and optimization method, the internal standards dissolved in the four solvents were tested in a mixture of 100 % methanol, methanol/water (50:50) (v/v), methanol + 0.01 % acetic acid, and ammonium buffer pH 3.5 [H 2 O/ACN (90:10) (v/v) + 1 mM NH 4 (CH 3 COO)]

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Two points deserve emphasis

Disrupt microbiome balance - Concussions can disrupt the balance of beneficial bacteria (12), allowing pro-inflammatory species to take over, contributing to increased inflammation

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