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MB Sample ID: SA176714

Local Sample ID:	P07B
Subject ID:	SU001984
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Subject:

Subject ID:	SU001984
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
P07B	SA176714	FL021901	Diabetes	Diagnosis

Collection:

Collection ID:	CO001977
Collection Summary:	Fasting plasma used for metabolomics was collected, kept at 4℃ in a freezer (CubeCooler®; Forte Grow Medical, Tochigi, Japan), and then subsequently frozen at −80℃
Sample Type:	Blood (plasma)

Treatment:

Treatment ID:	TR001996
Treatment Summary:	Systemically healthy controls and T2D patients were recruited.

Sample Preparation:

Sampleprep ID:	SP001990
Sampleprep Summary:	At first, 50 µL of plasma was mixed with 150 µL of deaerated H2O containing 0.2 mg/mL of ribitol, which was used as an internal standard, (1200 rpm, 10 min, 4°C). Then, 800 µL of deaerated MeCN was added (1200 rpm, 10 min, 4°C) and centrifuged (16000×g, 3 min, 4°C). Next, 300 µL of supernatant were transferred to an Eppendorf tube. All the steps up to this point were performed in a cold room at 4°C. The samples were then dried in a vacuum centrifuge dryer for 1 hour and lyophilized overnight. For derivatization, first, 100 µL of methoxyamine hydrochloride in pyridine (20 mg/mL) was added to the samples, and the mixture was incubated (1200 rpm, 90 min, 30°C). A second derivatizing agent, N-methyl-N-trimethysilyl-trifuroacetamide (MSTFA), was then added and the mixture was incubated (1200 rpm, 30 min, 37°C). After centrifucation (16000×g, 3 min), 100 µL of derivatized samples were transferred to glass vials.

Sampleprep ID:

SP001990

Sampleprep Summary:

At first, 50 µL of plasma was mixed with 150 µL of deaerated H2O containing 0.2 mg/mL of ribitol, which was used as an internal standard, (1200 rpm, 10 min, 4°C). Then, 800 µL of deaerated MeCN was added (1200 rpm, 10 min, 4°C) and centrifuged (16000×g, 3 min, 4°C). Next, 300 µL of supernatant were transferred to an Eppendorf tube. All the steps up to this point were performed in a cold room at 4°C. The samples were then dried in a vacuum centrifuge dryer for 1 hour and lyophilized overnight. For derivatization, first, 100 µL of methoxyamine hydrochloride in pyridine (20 mg/mL) was added to the samples, and the mixture was incubated (1200 rpm, 90 min, 30°C). A second derivatizing agent, N-methyl-N-trimethysilyl-trifuroacetamide (MSTFA), was then added and the mixture was incubated (1200 rpm, 30 min, 37°C). After centrifucation (16000×g, 3 min), 100 µL of derivatized samples were transferred to glass vials.

Combined analysis:

Analysis ID	AN003103
Analysis type	MS
Chromatography type	GC
Chromatography system	GC-MS/MS-TQ8040
Column	InertCap 5MS/NP capillary
MS Type	EI
MS instrument type	Triple quadrupole
MS instrument name	Shimazu TQ8040
Ion Mode	POSITIVE
Units	Intensity

Chromatography:

Chromatography ID:	CH002290
Instrument Name:	GC-MS/MS-TQ8040
Column Name:	InertCap 5MS/NP capillary
Chromatography Type:	GC

MS:

MS ID:	MS002885
Analysis ID:	AN003103
Instrument Name:	Shimazu TQ8040
Instrument Type:	Triple quadrupole
MS Type:	EI
MS Comments:	GC-MS data were converted into ABF format using an ABF converter, then processed using MS-DIAL (version 3.90) to perform feature detection, spectra deconvolution, metabolite identification, and peak alignment (Tsugawa et al. 2015). Normalization was then performed based on the internal standard (ribitol) as well as the LOWESS algorithm, whereby metabolic feature signal drift with time was independently corrected by fitting a LOWESS curve to the MS signal measured in QCs.
Ion Mode:	POSITIVE