Amino Acid Separation:Underivatized

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Amino Acid Separation without Derivatizing

This figure shows the extracted ion chromatogram (EIC) of a nineteen amino acid mixture under gradient conditions.

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Method Conditions and Sample Preparation Protocol

From our experience with amino acid separations in the ANP mode of separation, samples should be prepared as follows:

Stock sample solution (if used) should be prepared in 100% water with 0.1& formic acid (or acetic acid: depends which acid is used in your mobile phase).
Samples used for LC-MS should be diluted using 60% acetonitrile + 0.1%formic (or acetic acid) /40% DI water +0.1% formic (or acetic);

If the stock sample solution is not used sample should be prepared as in point 2 above.

Method Run Conditions:
Column: 2.1 x 150 mm Cogent Diamond Hydride™
Solvent A: DI water + 0.1% formic (or acetic) acid
Solvent B: Acetonitrile + 0.1% formic (or acetic acid)
Flow rate: 0.4 mL/min
Gradient conditions:
     0 min - 100 %B
     0-5 min to 75% B
     5 - 6 min to 60% B
     6 - 7 min hold 60% B
     7 - 9 min to 50%B
     9 - 10 min hold 50%B
     5 min post time at 100%B (for equlibration)
Please note Methanol is not a solvent of choice for ANP analysis, but Acetone is an excellent substitute when using LCMS.

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