Cheminformatics and statistical approaches were used to identify significant chemicals potentially associated with disease progression. Twelve Level 1-2a features were identified as statistically significant (Tukey's HSD post-hoc p-value < 0.05), summarized in Table and Significantly altered levels of valine and proline were observed, consistent with prior research indicating disrupted amino acids pathways in AD 11, , potentially due to the alteration of different neurotransmitters. Valine (Figure ) was significantly decreased in MCI and AD compared to the ND group, in line with previous studies reporting decreased valine levels in AD, associated with impaired neurotransmission and cognitive function . Furthermore, adenine (Figure ) was also significantly reduced in both MCI and AD, compared to ND. These results are consistent with previous studies performed in mice , indicating that the purine metabolism pathway may be altered in AD and potentially play an important role in the pathogenesis.