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MB Sample ID: SA031753
Local Sample ID: | END18_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). |