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MB Sample ID: SA031723
Local Sample ID: | C03_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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Subject:
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 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
C03_1 | SA031723 | FL007190 | Control | Sample Type |
Collection:
Collection ID: | CO000602 |
Collection Summary: | The FF samples were collected based on follicular aspiration after ovarian controlled stimulation protocol, defined by the doctor prior to the begining of the treatment. Follicular fluid samples were collectd and 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. |
Sample Type: | Follicular fluid |
Treatment:
Treatment ID: | TR000622 |
Treatment Summary: | Does not apply. Patients were treated for infertility according to their medical needs. The present study has no interference on patients management and it is defined as a prospective case-control study. |
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). |
Combined analysis:
Analysis ID | AN000900 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Ultima Q-TOF |
Column | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Waters Ultima QTOF |
Ion Mode | POSITIVE |
Units | unspecified |
Chromatography:
Chromatography ID: | CH000640 |
Instrument Name: | Ultima Q-TOF |
Column Name: | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) |
Column Temperature: | 35 °C |
Solvent A: | 100% water; 0.1% formic acid |
Solvent B: | 100% acetonitrile; 0.1% formic acid |
Analytical Time: | 17 minutes |
Capillary Voltage: | 3 kV |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS000802 |
Analysis ID: | AN000900 |
Instrument Name: | Waters Ultima QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
Ion Mode: | POSITIVE |