Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA138149

Local Sample ID:	2-A4-PS
Subject ID:	SU001709
Subject Type:	Mammal
Subject Species:	Rattus norvegicus
Taxonomy ID:	10116
Genotype Strain:	Sprague-Dawley
Age Or Age Range:	6 weeks-age
Weight Or Weight Range:	200 ± 10
Gender:	Male
Animal Animal Supplier:	Vital River Laboratories Co., Ltd (Beijng, China)
Animal Housing:	25 ± 2℃, 50±5% relative humidity
Animal Light Cycle:	12 h light/12 h dark photoperiod

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Subject:

Subject ID:	SU001709
Subject Type:	Mammal
Subject Species:	Rattus norvegicus
Taxonomy ID:	10116
Genotype Strain:	Sprague-Dawley
Age Or Age Range:	6 weeks-age
Weight Or Weight Range:	200 ± 10
Gender:	Male
Animal Animal Supplier:	Vital River Laboratories Co., Ltd (Beijng, China)
Animal Housing:	25 ± 2℃, 50±5% relative humidity
Animal Light Cycle:	12 h light/12 h dark photoperiod

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
2-A4-PS	SA138149	FL017570	Model	class
2-A4-PS	SA138149	FL017570	plasma	Tissue

Collection:

Collection ID:	CO001702
Collection Summary:	Blood samples of the rats were collected to obtain plasma by centrifugation (3000r/min, 15 min) using heparin as an anticoagulant and kept at -80℃ until analysis. Hearts were collected, washed with cold saline and cut in half. One part was grinded into powder in liquid nitrogen and kept at -80℃. Heart samples were homogenized in saline on ice at the ratio of tissue to solution was 1:4 (w/v) before analysis.
Sample Type:	Heart

Treatment:

Treatment ID:	TR001722
Treatment Summary:	Rats were divided into six rats in each group randomly as following: control rats group, MI model rats group, MI model rats pre-treated with diltiazem group (50 mg/kg, i.g.), and MI model rats pre-treated with GTSS group (200 mg/kg, i.g.). Pre-treated rats were consecutively administrated for 4 weeks. Except control rats group received saline, other rat groups received two injections of isoproterenol (85 mg/kg, s.c.) on the 27th and 28th days at an interval of 24 hours.

Treatment ID:

TR001722

Treatment Summary:

Rats were divided into six rats in each group randomly as following: control rats group, MI model rats group, MI model rats pre-treated with diltiazem group (50 mg/kg, i.g.), and MI model rats pre-treated with GTSS group (200 mg/kg, i.g.). Pre-treated rats were consecutively administrated for 4 weeks. Except control rats group received saline, other rat groups received two injections of isoproterenol (85 mg/kg, s.c.) on the 27th and 28th days at an interval of 24 hours.

Sample Preparation:

Sampleprep ID:	SP001715
Sampleprep Summary:	100μl of plasma or tissue homogenate was transferred into a 10 ml glass tube and added 20 μl of lipid internal standards. After vortexed for 10 s, 1.5 ml of methanol and 5 ml methyl tert-butyl ether were added to the tube, and the sample was vortexed for 15 min and then added 1.5 ml water, then centrifuged at 4500r/min for 10 min (4 ℃). The upper organic phase was transferred to another glass tube, and the lower water phase was added with 2 ml secondary extractant (methyl tert-butyl ether /methanol/water,10:3:2.5, v/v/v). After vortexed for 10min and centrifuged for 15min at 4500rpm, and the upper organic phase was combined and dried under a gentle nitrogen stream. The residue was re-dissolved in 100 μl methanol/chloroform (1:1, v/v).

Sampleprep ID:

SP001715

Sampleprep Summary:

100μl of plasma or tissue homogenate was transferred into a 10 ml glass tube and added 20 μl of lipid internal standards. After vortexed for 10 s, 1.5 ml of methanol and 5 ml methyl tert-butyl ether were added to the tube, and the sample was vortexed for 15 min and then added 1.5 ml water, then centrifuged at 4500r/min for 10 min (4 ℃). The upper organic phase was transferred to another glass tube, and the lower water phase was added with 2 ml secondary extractant (methyl tert-butyl ether /methanol/water,10:3:2.5, v/v/v). After vortexed for 10min and centrifuged for 15min at 4500rpm, and the upper organic phase was combined and dried under a gentle nitrogen stream. The residue was re-dissolved in 100 μl methanol/chloroform (1:1, v/v).

Combined analysis:

Analysis ID	AN002667	AN002668
Analysis type	MS	MS
Chromatography type	Reversed phase	Reversed phase
Chromatography system	Agilent 1100	Thermo Accela 1250
Column	Peeke C8 SR (150×3.0mm, 3μm)	Waters Xterra MS C8 (100 x 2.1mm,3.5um)
MS Type	ESI	ESI
MS instrument type	Triple quadrupole	LTQ-FT
MS instrument name	Agilent 6410 QQQ	Thermo LTQ-FT
Ion Mode	POSITIVE	UNSPECIFIED
Units	pmol/ml(PL) & pmol/mg(HL)	pmol/ml(PS) & pmol/mg(HS)

Chromatography:

Chromatography ID:	CH001962
Instrument Name:	Agilent 1100
Column Name:	Peeke C8 SR (150×3.0mm, 3μm)
Chromatography Type:	Reversed phase

Chromatography ID:	CH001963
Instrument Name:	Thermo Accela 1250
Column Name:	Waters Xterra MS C8 (100 x 2.1mm,3.5um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS002466
Analysis ID:	AN002667
Instrument Name:	Agilent 6410 QQQ
Instrument Type:	Triple quadrupole
MS Type:	ESI
MS Comments:	The parameters for electrospray ionization tandem MS in positive ion mode were as follows: gas temperature, 350 ℃; gas flow rate, 10 l/min; nebulizer, 30 psi; and capillary voltage, 4000V. Multiple reaction monitoring was performed using the characteristic precursor-to-product ion transitions, optimized fragmentor voltages, and collision energies.
Ion Mode:	POSITIVE

MS ID:	MS002467
Analysis ID:	AN002668
Instrument Name:	Thermo LTQ-FT
Instrument Type:	LTQ-FT
MS Type:	ESI
MS Comments:	The LTQ-FT was run in full-scan mode at 100000 resolution from m/z50–1200 with MS parameters: sheath gas flow rate, 50 arb; aux gas flow rate, 20 arb; sweep gas flow rate, 3 arb; capillary temperature, 275 ℃ in both positive and negative mode. And spray voltage, 4.5kV; capillary voltage, 35.0V; tube lens, 120V were used for the positive mode. Spray voltage, -4.0kV; capillary voltage, -35.0V; tube lens, -120V were used for the negative mode.
Ion Mode:	UNSPECIFIED