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Blood samples were obtained when the patients were diagnosed with NTM-PD or BE during their routine health check-ups. The collected samples were incubated at 25°C for 15–30 min and centrifuged at 13,756 × g for 20 min to isolate serum. The serum samples (500 μL) were stored at −80°C until further analysis. Before metabolomic analysis, the frozen samples were thawed on ice and vortexed. Then, each serum (80 μL) was deproteinized by adding 350 μL prechilled acetonitrile/methanol (1:1, v/v), followed by shaking for 5 min and centrifugation at 18,341 × g for 10 min. Supernatant (400 μL) was transferred to a new tube and then further centrifuged. The resulting supernatant (350 μL) was diluted with equal volume of distilled water, and 150 μL of the final solution was transferred to an autosampler vial for analysis. A pooled quality control (PQC) sample was created by combining 50 μL aliquots from all serum samples. The PQC sample was randomly placed within the sequence after preparation, alongside the analysis samples.

Sample separation was performed on ACQUITY UPLC HSS T3 1.8 μM (2.1 × 100 mm) at a temperature of 50℃. The mobile phase consisted of 0.1% formic acid in water (A) and 0.1% formic acid in methanol (B). The gradient program initiated with 5% B for 1 min, 5%–95% B for 1–6 min, 95%–98% for 6–12 min, stayed at 98% B for 12–22 min, and equilibrated to the initial condition for 22–25 min. , Sample separation was performed on ACQUITY UPLC HSS T3 1.8 μM (2.1 × 100 mm) at a temperature of 50℃. The mobile phase consisted of 0.1% formic acid in water (A) and 0.1% formic acid in methanol (B). The gradient program initiated with 5% B for 1 min, 5%–95% B for 1–6 min, 95%–98% for 6–12 min, stayed at 98% B for 12–22 min, and equilibrated to the initial condition for 22–25 min.

The mass scan range was 66.7–1000 m/z in the full scan mode. Data were acquired with a mass resolution of 70,000 full width at half maximum (FWHM) at m/z 200, injection time of 100 ms, and automatic gain control target 1e6. , The mass scan range was 66.7–1000 m/z in the full scan mode. Data were acquired with a mass resolution of 70,000 full width at half maximum (FWHM) at m/z 200, injection time of 100 ms, and automatic gain control target 1e6.