Proline: Trans-4-Hydroxy-L-Proline by LCMS – AppNote

Rapid, High Efficiency Method by LCMS

This method is highly specific, efficient and fast for the analysis detection, small volumes of the reaction mixture can be injected and of trans-4-hydroxy-L-proline. Due to the high specificity of the the amount of the produced compound can be determined. No derivatization is required for the detection of this important compound.

Trans-4-Hydroxy-L-Proline 132.0655 m/z (M + H)+

Method Conditions
Column: Cogent Diamond Hydride™, 4μm, 100Å
Catalog No.: 70000-05P-2
Dimensions: 2.1 x 50mm
Mobile Phase:
A: 50% Methanol / 50% DI Water / 0.05% Acetic Acid
B: 97% Acetonitrile / 3% DI Water / 0.05% Acetic Acid

Time (minutes) %B
0 95
10 30
11 30
12 95

Post Time: 5 minutes
Flow rate: 0.4 mL / minute
Detection: ESI – POS – Agilent 6210 MSD TOF Mass Spectrometer
Injection vol.: 1μL
Sample Preparation:
Stock Solution: 1 mg / mL in Methanol diluent.
Working Solution: Stock aliquot was diluted using 50% Solvent A and 50% Solvent B mixture for the final concentration 0.5 mg / L. Before injection, solution was filtered using a 0.45µm Nylon Syringe Filter (MicroSolv Tech Corp.).

Note: Isomers of Hydroxyproline have been found in nature. Trans-4-Hydroxy-L-Proline is the major component of Collagen, Gelatin, plant wall Proteins, etc. It is a useful material for synthesis of pharmaceuticals such as Angiotensin converting enzyme inhibitors and Carbapenem antibiotics.


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