Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA156588

Local Sample ID:	792 _2
Subject ID:	SU001766
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Gender:	Not applicable

Select appropriate tab below to view additional metadata details:

Subject:

Subject ID:	SU001766
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Gender:	Not applicable

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
792 _2	SA156588	FL018359	3 months	Sample Code

Collection:

Collection ID:	CO001759
Collection Summary:	Breast milk samples were collected at room temperature and stored at −80 °C until analyzed.
Sample Type:	Breast milk

Treatment:

Treatment ID:	TR001779
Treatment Summary:	Sample were stored at -80 as it is, no treatment done.

Sample Preparation:

Sampleprep ID:	SP001772
Sampleprep Summary:	30 µl breast milk sample or 10 µl of serum of was extracted with using a modified version of the previously published Folch procedure (Nygren et al. 2011). In short, 10 µL of 0.9% NaCl and, 120 µL of CHCl3: MeOH (2:1, v/v) containing the internal standards (c = 2.5 µg/mL) was added to the sample. The standard solution contained the following compounds: 1,2-diheptadecanoyl-sn-glycero-3-phosphoethanolamine (PE(17:0/17:0)), N-heptadecanoyl-D-erythro-sphingosylphosphorylcholine (SM(d18:1/17:0)), N-heptadecanoyl-D-erythro-sphingosine (Cer(d18:1/17:0)), 1,2-diheptadecanoyl-sn-glycero-3-phosphocholine (PC(17:0/17:0)), 1-heptadecanoyl-2-hydroxy-sn-glycero-3-phosphocholine (LPC(17:0)) and 1-palmitoyl-d31-2-oleoyl-sn-glycero-3-phosphocholine (PC(16:0/d31/18:1)), were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL, USA), and, triheptadecanoylglycerol (TG(17:0/17:0/17:0)) was purchased from Larodan AB (Solna, Sweden). The samples were vortex mixed and incubated on ice for 30 min after which they were centrifuged (9400 × g, 3 min). 60 µL from the lower layer of each sample was then transferred to a glass vial with an insert and 60 µL of CHCl3: MeOH (2:1, v/v) was added to each sample. The samples were stored at -80 °C until analysis.

Sampleprep ID:

SP001772

Sampleprep Summary:

30 µl breast milk sample or 10 µl of serum of was extracted with using a modified version of the previously published Folch procedure (Nygren et al. 2011). In short, 10 µL of 0.9% NaCl and, 120 µL of CHCl3: MeOH (2:1, v/v) containing the internal standards (c = 2.5 µg/mL) was added to the sample. The standard solution contained the following compounds: 1,2-diheptadecanoyl-sn-glycero-3-phosphoethanolamine (PE(17:0/17:0)), N-heptadecanoyl-D-erythro-sphingosylphosphorylcholine (SM(d18:1/17:0)), N-heptadecanoyl-D-erythro-sphingosine (Cer(d18:1/17:0)), 1,2-diheptadecanoyl-sn-glycero-3-phosphocholine (PC(17:0/17:0)), 1-heptadecanoyl-2-hydroxy-sn-glycero-3-phosphocholine (LPC(17:0)) and 1-palmitoyl-d31-2-oleoyl-sn-glycero-3-phosphocholine (PC(16:0/d31/18:1)), were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL, USA), and, triheptadecanoylglycerol (TG(17:0/17:0/17:0)) was purchased from Larodan AB (Solna, Sweden). The samples were vortex mixed and incubated on ice for 30 min after which they were centrifuged (9400 × g, 3 min). 60 µL from the lower layer of each sample was then transferred to a glass vial with an insert and 60 µL of CHCl3: MeOH (2:1, v/v) was added to each sample. The samples were stored at -80 °C until analysis.

Combined analysis:

Analysis ID	AN002759
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Waters Acquity UPLC
Column	Waters Acquity CSH C18 (100 x 2.1mm,1.7um)
MS Type	ESI
MS instrument type	QTOF
MS instrument name	Agilent 6210 TOF
Ion Mode	NEGATIVE
Units	ng/ml

Chromatography:

Chromatography ID:	CH002039
Chromatography Summary:	BEH C18 (2.1 x 100 mm, particle size 1.7 µm) (Waters Corporation, Milford, MA, USA)
Instrument Name:	Waters Acquity UPLC
Column Name:	Waters Acquity CSH C18 (100 x 2.1mm,1.7um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS002556
Analysis ID:	AN002759
Instrument Name:	Agilent 6210 TOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	Dual jet stream electrospray (dual ESI) ion source was used and the ion polarity was on negative mode. The capillary voltage and the nozzle voltage were kept at 4500 V and 1500 V. The N2 pressure was set on 21 psi, with the sheath gas flow as 11 L/min and temperature at 379°C for the nebulizer. The data was acquired with MassHunter B.06.01 software (Agilent Technologies, Santa Clara, CA, The United States of America).
Ion Mode:	NEGATIVE