Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA031752

Local Sample ID:	END08_1
Subject ID:	SU000608
Subject Type:	Human Follicular Fluid
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Age Or Age Range:	18-40
Gender:	female
Human Medications:	Controled ovarian Stimulation
Human Exclusion Criteria:	Polycystic ovary syndrome, cancer, premature ovarian failure.
Species Group:	Human

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Sample Preparation:

Sampleprep ID:	SP000615
Sampleprep Summary:	The FF samples were centrifuged at 800 × g for 10 minutes to separate the fluid from follicular cells and the samples were maintained frozen in -20 °C until the metabolites extraction. The Bligh & Dyer protocol (1959) was applied for proteins removal and was performed as herein: 50 µL of sample was placed in a microtube, followed by addition of 50 µL of milli-Q water, 125 µL of chloroform (CHCl3 - Merck Millipore, MA, EUA) and 250 µL of methanol (MeOH - Merck Millipore - Massachusetts, EUA). The mixture was vortexed for 30 seconds. Next, polar and apolar phases were separated by the addition of 100 µL of milli-Q water and 125 µL of CHCl3 (Merck Millipore, MA EUA). Both polar and apolar phases were collected and transferred to a new microtube. The final content was dried by using a speedvac (CentriVap Cold Trap, Labconco, MO, USA). Prior to the LC-MS analysis, the metabolites were recovered by using 100 µL of acetonitrile and water (ACN:H2O 1:1, v:v - Merk Millipore, MA, USA) and filtered in 0.22 μm PVDF filters (Merck Millipore, MA, EUA).

Sampleprep ID:

SP000615

Sampleprep Summary:

The FF samples were centrifuged at 800 × g for 10 minutes to separate the fluid from follicular cells and the samples were maintained frozen in -20 °C until the metabolites extraction. The Bligh & Dyer protocol (1959) was applied for proteins removal and was performed as herein: 50 µL of sample was placed in a microtube, followed by addition of 50 µL of milli-Q water, 125 µL of chloroform (CHCl3 - Merck Millipore, MA, EUA) and 250 µL of methanol (MeOH - Merck Millipore - Massachusetts, EUA). The mixture was vortexed for 30 seconds. Next, polar and apolar phases were separated by the addition of 100 µL of milli-Q water and 125 µL of CHCl3 (Merck Millipore, MA EUA). Both polar and apolar phases were collected and transferred to a new microtube. The final content was dried by using a speedvac (CentriVap Cold Trap, Labconco, MO, USA). Prior to the LC-MS analysis, the metabolites were recovered by using 100 µL of acetonitrile and water (ACN:H2O 1:1, v:v - Merk Millipore, MA, USA) and filtered in 0.22 μm PVDF filters (Merck Millipore, MA, EUA).