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LC/MS Analysis of L-Methionine Sulfoximine, Sulfone and Sulfoxide on Ascentis® Express HILIC

LC/MS Analysis of L-Methionine Sulfoximine, Sulfone and Sulfoxide on Ascentis® Express HILIC application for HPLC

Materials

Analytical column

Product No.
Description
Pricing

Ascentis® Express HILIC, 2.7 μm HPLC Column

2.7 μm particle size, L × I.D. 10 cm × 3 mm

Standard

Product No.
Description
Pricing

L-Methionine sulfone

L-Methionine sulfoxide

L-Methionine sulfoximine

suitable for cell culture

CONDITIONS

column

Ascentis Express HILIC, 10 cm x 3.0 mm I.D., 2.7 μm particles (53970-U)

mobile phase

[A] acetonitrile; [B] water with 0.1% formic acid, pH 3.5 with ammonium formate; (75:25, A:B)

flow rate

0.4 mL/min

pressure

1350 psi 93 bar

temp.

25 °C

detector

MS, ESI (+), TIC, m/z 150-300

injection

5 μL

sample

10 μg/mL in 90:10, methanol:water

Description

Analysis Note

L-methionine sulfoximine and L-buthionine sulfoximine are used in are used to prevent additional enzyme activity in Chinese hampster ovary (CHO) cell lines with supplemental Glutamine Synthase (GS). It is of interest to be able to monitor both purity of such molecules in order to control addition to cell media as well as for the assay of parent molecules during use. The analytes are highly polar and thus should be amenable to HILIC-LC-MS analysis. This first installment investigates the ability of Ascentis Express HILIC and Ascentis Express OH5 stationary phases for the separation of the methionine analog and related compounds as well as methionine and buthionine separations.

Legal Information

Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany