Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA206257

Local Sample ID:	Q13_Positive_PostspikeB_Positive_1
Subject ID:	SU002237
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Sample Preparation:

Sampleprep ID:	SP002243
Sampleprep Summary:	One sample per tissue was used in chemical analyses. Chemicals were separated from tissue via solid-liquid extraction by adding 2 mL of room temperature water:acetonitrile (1:1) per sample. Placenta samples were disrupted and homogenized using a TissueRuptor (Qiagen), vortexed for 5 min, and placed in a centrifuge at 4000 rpm for 5 min. The resulting supernatant was collected, and a second round of solid-liquid extraction was performed using 8 mL acetonitrile with 2% formic acid. Samples were vortexed and centrifuged again, and both supernatants combined yielding a final total volume of 10 mL. Of the 10 mL combined supernatant sample, 3 mL were then run through solid filtration with a Captiva EMR-lipid column (Agilent Technologies). Resulting eluates were placed into liquid chromatography mass spectrometry (LCMS) vials in 1.0 mL aliquots. Remaining supernatant volumes were banked at -80°C. Method blanks were collected in parallel using the same steps without adding tissues. For non-targeted chemical analysis, 20 µL of 1.25 µg/mL tracer solution was added to each 1.0 mL aliquot of post-filtration eluate (for study samples, blanks, and controls) yielding a final concentration of 25 ng/mL per sample. In addition, tracers were added to a single sample in duplicate prior to extraction to evaluate extraction recovery and reproducibility.

Sampleprep ID:

SP002243

Sampleprep Summary:

One sample per tissue was used in chemical analyses. Chemicals were separated from tissue via solid-liquid extraction by adding 2 mL of room temperature water:acetonitrile (1:1) per sample. Placenta samples were disrupted and homogenized using a TissueRuptor (Qiagen), vortexed for 5 min, and placed in a centrifuge at 4000 rpm for 5 min. The resulting supernatant was collected, and a second round of solid-liquid extraction was performed using 8 mL acetonitrile with 2% formic acid. Samples were vortexed and centrifuged again, and both supernatants combined yielding a final total volume of 10 mL. Of the 10 mL combined supernatant sample, 3 mL were then run through solid filtration with a Captiva EMR-lipid column (Agilent Technologies). Resulting eluates were placed into liquid chromatography mass spectrometry (LCMS) vials in 1.0 mL aliquots. Remaining supernatant volumes were banked at -80°C. Method blanks were collected in parallel using the same steps without adding tissues. For non-targeted chemical analysis, 20 µL of 1.25 µg/mL tracer solution was added to each 1.0 mL aliquot of post-filtration eluate (for study samples, blanks, and controls) yielding a final concentration of 25 ng/mL per sample. In addition, tracers were added to a single sample in duplicate prior to extraction to evaluate extraction recovery and reproducibility.