| Summary: | 碩士 === 輔仁大學 === 化學系 === 106 === Recently, many studies shown alteration of tryptophan (Trp) metabolism elicited by proinflammatory cytokines gained more attention as a new notion were associated with several neurological and inflammatory disorders such as patients with primary Sjogren’s syndrome, lung cancer and Alzheimer disease.
The sample preparation methods included PMME ( Polymer monolith microextraction) and d-SPE (Dispersive solid phase extraction) . The sample preparation methods were applied for Trp metabolites, including xanthurenic acid (XANA), tryptophan (Trp), serotonin (5-HT), 5-hydroxy-indole-3-acetic acid (5-HIAA), kynurenine (KYN), kynurenic acid (KYNA), 3-hydroxyanthranilic acid (3-HAA), anthranilic acid (AA), 3-hydroxykynurenine (3-HK), and quinolinic acid (QA) by UPLC-DAD ( Ultra performance liquid chromatography- Diode array detection) and LC-MS/MS (Liquid chromatography tandem mass spectrometry).
The PMME optimal parameters, loading speed for the PMME was 2.5 mL/hr and the elute solvent was 100 μL MeOH and 100 μL MeOH:NH4OH (95:5,v/v). The coefficients of correlation (r2) for the UPLC-DAD calibration curves were greater than 0.9995. The limit of detection (LOD) and limit of quantification (LOQ) were determined within 1.56 to 19.82 ng/mL and 5.20 to 66.06 ng/mL.The recoverys of tryptophan metabolites in spike serum were 70.67 % to 132.67 %, standard deviation range from 1.33 to 12.98.
Under optimal conditions, the d-SPE methods were obtained such as extraction time of 1 min, 5 mg of PHF-1a, 400 μL of MeOH as desorption solvent and desorption time of 3 min. The coefficients of correlation (r2) for the LC-MS/MS calibration curves were greater than 0.9973. The limit of detection (LOD) and limit of quantification (LOQ) were determined within 0.002 to 0.127 ng/mL and 0.008 to 0.424 ng/mL.The recoverys of tryptophan metabolites in spike serum were 77.13 % to 113.30 %, standard deviation range from 0.60 to 18.48. The intra-day and inter-day standard deviation range from 0.63 % to 19.52 % and 5.24 % to 18.29 %. Satisfactory results shown that two pretreament methods could be simple and rapid extraction of tryptophan metabolites in human serum serum.
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