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MB Sample ID: SA022647
| Local Sample ID: | UNIFESP_lip_LF18 |
| Subject ID: | SU000466 |
| Subject Type: | Human patients (Women) |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | <= 37 years |
| Gender: | Female |
| Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000466 |
| Subject Type: | Human patients (Women) |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | <= 37 years |
| Gender: | Female |
| Species Group: | Human |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| UNIFESP_lip_LF18 | SA022647 | FL005551 | LH | Treatment |
Collection:
| Collection ID: | CO000460 |
| Collection Summary: | The remaining follicular fluid from follicles aspiration (approximately 15 mL/patient) was centrifuged at 800 x g for 10 minutes to remove residual cells, and the supernatant was collected and stored at -20 °C until lipid analysis. |
| Sample Type: | Follicular Fluid |
| Collection Method: | Ovarian pucture |
| Collection Location: | Ovarian follicle |
| Collection Frequency: | One time |
Treatment:
| Treatment ID: | TR000480 |
| Treatment Summary: | To induce follicular growth, each of the eligible 14 patients received both rFSH 225 IU alone (Gonal F® [Merck-Serono, Darmstadt, Germany]) (FSH group) and rFSH 225 IU + hMG 75 IU (Menopur® [Ferring, Kiel, Germany]) (Low-dose-LH group) starting on cycle day 2 in two separate cycles (i.e. each patient acted as her own control). Sequential transvaginal ultrasounds were performed from stimulation day 5 onwards. Daily doses of GnRH antagonist analog cetrorelix acetate 0.25 mg (Cetrotide® [Merck-Serono, Darmstadt, Germany]) were administered in both groups when the leading follicle reached a mean diameter of 13 mm until the stimulation day 7, or from day 8. When at least one follicle was greater than 17 mm, 250 mcg of human chorionic gonadotropin (hCG) (Ovidrel® [Merck-Serono, Darmstadt, Germany]) was administered, and oocyte retrieval was scheduled 35-36 hours later. |
| Treatment: | Two types of controlled ovarian stimulation |
| Treatment Compound: | Exogenous gonadotropins to induce follicular growth |
Sample Preparation:
| Sampleprep ID: | SP000473 |
| Sampleprep Summary: | After thawing, lipids and metabolites were extracted based on the Bligh and Dyer protocol [26]. Briefly, 50 µL of sample was placed in a microtube, and 125 μL of chloroform (CHCl3) (Merck-Serono, Darmstadt, Germany) and 250 μL of methanol (CH3OH) (Merck-Serono, Darmstadt, Germany) were added. The mixture was submitted to homogenization for 2 minutes. The polar and nonpolar phases were separated by the addition of 125 μL of CHCl3 and 100 μL of Milli-Q water. Then, the samples were centrifuged for 5 minutes at 1000 x g. The organic phase (bottom) containing the lipids was recovered and transferred to a clean microtube, which was left open overnight at room temperature until the complete solvent evaporation. |
| Extraction Method: | Modified Bligh and Dyer protocol |
Combined analysis:
| Analysis ID | AN000696 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Waters Acquity I-Class |
| Column | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Waters Synapt G2 S QTOF |
| Ion Mode | POSITIVE |
| Units | m/z intensity |
Chromatography:
| Chromatography ID: | CH000504 |
| Chromatography Summary: | Reversed-phase analysis was performed on a Waters Ultra Performance liquid chromatography Acquity I-Class system using an Acquity HSS T3 2.10 x 100 mm column (Waters, Milford, USA). The column was maintained at 55 °C. The lipid extracts were analyzed by UPLC-MSE by injecting 10 μl of each sample dissolved in 1 mL of water:acetonitrile:2-propanol (H2O:ACN:2-propanol; 1:1:2, v:v:v) (Merck, Darmstadt, Germany). The chromatographic run was carried out in gradient mode with a flow rate of 0.5 mL min-1 with initial composition of 50% solvent B (ACN:2-propanol; 10:90) in solvent A (H2O:ACN; 40:60, v:v + 10 mM ammonium acetate (NH4Ac) (Sigma-Aldrich, Saint Louis, USA), maintained for 0 to 0.5 min. The gradient composition was changed linearly from 50% to 100% solvent B between 0.5 to 4.5 minutes, returning to the initial composition of 50% B in 4.6 minutes, kept to 5.0 minutes. |
| Instrument Name: | Waters Acquity I-Class |
| Column Name: | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) |
| Column Temperature: | 55ºC |
| Flow Rate: | 0.5 mL.min-1 |
| Solvent A: | 40% water/60% acetonitrile; 10 mM ammonium acetate |
| Solvent B: | 10% acetonitrile/90% isopropanol |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000618 |
| Analysis ID: | AN000696 |
| Instrument Name: | Waters Synapt G2 S QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
