研究目的
To understand why other studies failed to detect Trp after CBQCA derivatization, by studying the Trp derivatization using MS, by comparing ?uorescence intensity of Trp and Leu after derivatization and by using (cid:3)-cyclodextrin in the buffer as an enhancement factor of the ?uorescence yield.
研究成果
The study demonstrated that Trp can be identified using CBQCA derivatization and CE/LEDIF, despite its low fluorescence yield. The addition of (cid:3)-CD improved the sensitivity of Trp-CBQCA fluorescence detection. The decarboxylation observed during MS analysis occurs in the source of the mass spectrometer, not during the derivatization process.
研究不足
The main limitation is the poor fluorescent yield of the Trp-CBQCA derivative, which makes its identification challenging. Additionally, the decarboxylation of the labeled amino acids in the mass spectrometer source complicates the analysis.
1：Experimental Design and Method Selection:
The study involved CE/LEDIF experiments and MS analysis to understand the derivatization and fluorescence properties of Trp-CBQCA and Leu-CBQCA.
2：Sample Selection and Data Sources:
Samples of Trp and Leu were derivatized with CBQCA under specific conditions.
3：List of Experimental Equipment and Materials:
An Agilent Technologies CE7100 instrument, a Zetalif LED detector, and a Micromass Qtof Ultima 2 mass spectrometer were used. Chemicals included boric acid, SDS, gamma-cyclodextrin, beta-cyclodextrin, L-Leu, DL-Trp, nonylamine, KCN, and CBQCA.
4：Experimental Procedures and Operational Workflow:
Derivatization was performed in borate buffer at pH
5：9, followed by CE/LEDIF and MS analysis. Data Analysis Methods:
Fluorescence and absorbance measurements were conducted, and MS/MS spectra were analyzed to understand the derivatization process.
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