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The elution sequence, for the digested protein samples, included a linear gradient from 10% to 60% of eluent B for 52 min, then a plateau at 95 % of eluent B for 4 min

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Although, zopolrestat and tolrestat were discontinued due to severe hepatotoxicity and an unfavorable risk-benefit profile observed in phase 2 and phase 3 clinical trials, respectively [150152], and ranirestat, which also progressed through phase 3 trial, focused primarily on diabetic polyneuropathy rather than cardiovascular outcomes [153]