Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA031774

Local Sample ID:	QC_Control_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
QC_Control_1	SA031774	FL007192	Quality 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).

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