Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA301654

Local Sample ID:	837P
Subject ID:	SU002918
Subject Type:	Cultured cells
Subject Species:	Lentilactobacillus kefiri

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Sample Preparation:

Sampleprep ID:	SP002924
Sampleprep Summary:	Cell pellets were washed 3 times with cold PBS to remove any cell media left after collection. The cell pellets were then extracted using cold 100% methanol and spiked with 40 μL of 10 pinitol (internal standard). A quality control (QC) sample was prepared by mixing the same amount of each sample into one. The supernatants were then dried down using a speed-vac and then resuspended in 20 mg/mL methoxyamine hydrochloride reagent prepared in pure pyridine and incubated for 2 hr at 37 °C on a platform shaker at 1000 rpm. Next, for derivatization, the MSTFA +1% TMCS deri-vatization (ThermoFisher) was added to each sample, incubated for 30 min at 37°C on a platform shaker at 1000 rpm followed by a centrifugation for 10 min at 16,000 g prior to transferring the mixture to GC vials for injection into GC-MS.

Sampleprep ID:

SP002924

Sampleprep Summary:

Cell pellets were washed 3 times with cold PBS to remove any cell media left after collection. The cell pellets were then extracted using cold 100% methanol and spiked with 40 μL of 10 pinitol (internal standard). A quality control (QC) sample was prepared by mixing the same amount of each sample into one. The supernatants were then dried down using a speed-vac and then resuspended in 20 mg/mL methoxyamine hydrochloride reagent prepared in pure pyridine and incubated for 2 hr at 37 °C on a platform shaker at 1000 rpm. Next, for derivatization, the MSTFA +1% TMCS deri-vatization (ThermoFisher) was added to each sample, incubated for 30 min at 37°C on a platform shaker at 1000 rpm followed by a centrifugation for 10 min at 16,000 g prior to transferring the mixture to GC vials for injection into GC-MS.