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MB Sample ID: SA333604
| Local Sample ID: | old_9 |
| Subject ID: | SU003219 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | Young donors (aged 25 years or younger) and older donors (aged 50 years or older). |
| Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003219 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | Young donors (aged 25 years or younger) and older donors (aged 50 years or older). |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| old_9 | SA333604 | FL039611 | heart tissue | Sample source |
| old_9 | SA333604 | FL039611 | Old | Group |
| old_9 | SA333604 | FL039611 | Male | Sex |
Collection:
| Collection ID: | CO003212 |
| Collection Summary: | Control donor hearts that were considered suitable for transplantation but ultimately not used due to factors such as transport issues, immune mismatches, and size discrepancies between donor and recipient, rather than being post-mortem specimens were collected in this study. These hearts came from individuals who died of non-cardiac reasons and had no risk factors for heart disease, including having a BMI under 30. Following a thorough pathological review, these hearts were confirmed to be structurally normal through histological evaluation. Left ventricular (LV) samples were taken directly in the operating room, immediately snap-frozen in liquid nitrogen at -196°C, and then stored at the Sydney Heart Bank at the University of Sydney, at temperatures ranging from -170 to -180°C. This collection process received the ethical green light from the University of Sydney's Ethics Committee (USYD # 2021/122). |
| Sample Type: | Heart |
| Storage Conditions: | Described in summary |
Treatment:
| Treatment ID: | TR003228 |
| Treatment Summary: | The samples detailed in this study did not undergo any treatment. |
Sample Preparation:
| Sampleprep ID: | SP003225 |
| Sampleprep Summary: | Briefly, approximately 50 mg of ground heart tissue was weighed into 2 mL Eppendorf tube and subjected to a two-phase extraction protocol involving tissue, ice-cold extraction medium (methanol:water, and chloroform). Samples were then centrifuged at 14 300 rpm at 4 °C for 25 min. The aqueous layer was transferred into a new microfuge tube, concentrated in the Speed-Vac SPD120 (Thermo Fisher Scientific) and dried under nitrogen stream, followed by reconstitution in the acetonitrile/methanol/formic acid (75:25:0.2; v/v/v, HPLC grade; Thermo Fisher Scientific) for the HILIC analysis, and acetonitrile/methanol (25:25; v/v/v, HPLC grade) for the AMIDE analysis. |
Combined analysis:
| Analysis ID | AN005082 | AN005083 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Agilent 1260 | Agilent 1260 |
| Column | Waters Atlantis T3 (150 x 4.6mm,5um) | Waters XBridge BEH Amide (100 x 2.1mm,2.5um) |
| MS Type | API | API |
| MS instrument type | Triple quadrupole | Triple quadrupole |
| MS instrument name | ABI Sciex 5500 QTrap | ABI Sciex 5500 QTrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | abundance | abundance |
Chromatography:
| Chromatography ID: | CH003839 |
| Chromatography Summary: | Chromatographic method to detect analytes such as amino acids, endocannabinoids, fatty acid β-oxidation intermediates, gut-derived metabolites, nucleotides, neurotransmitters, cofactors and vitamins. |
| Instrument Name: | Agilent 1260 |
| Column Name: | Waters Atlantis T3 (150 x 4.6mm,5um) |
| Column Temperature: | 40 |
| Flow Gradient: | The LC gradient program begins with an initial condition of 5% solvent A and 95% solvent B, with a flow rate of 0.25 mL/min, which is held for 0.5 minutes to establish system equilibration. The gradient then proceeds as follows: at 6 minutes, the mobile phase composition shifts to 60% solvent A and 40% solvent B for 3minutes; at 10 minutes, it changes back to 5% solvent A and 95% solvent B; at 11 minutes, the flow rate increases to 0.4 mL/min while maintaining a composition of 5% solvent A and 95% solvent B for a duration of 12.5 minutes; and at 24.5 minutes, the flow rate decreases back to 0.25 mL/min. The final condition is maintained for 1 minutes to ensure stability before subsequent analyses. |
| Flow Rate: | 0.250 – 0.400 mL/min |
| Solvent A: | 100% water; 0.1% formic acid; 10 mM ammonium formate (pH ~2.5) |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH003840 |
| Chromatography Summary: | Chromatographic method to detect analytes including amino acids, nucleotides, nucleosides, nucleotide triphosphates, high energy intermediates, organic acids, TCA cycle intermediates, bile acids and vitamins. |
| Instrument Name: | Agilent 1260 |
| Column Name: | Waters XBridge BEH Amide (100 x 2.1mm,2.5um) |
| Column Temperature: | 40 |
| Flow Gradient: | The LC gradient program is initialized with an initial mobile phase composition of 15% solvent A and 85% solvent B at a flow rate of 0.25 mL/min. Over the course of 8 minutes, the mobile phase composition undergoes a transition to 65% solvent A and 35% solvent B. Subsequently, at 8 minutes, the composition shifts to 98% solvent A and 2% solvent B, maintained for 1 minute. The mobile phase reverts back to the initial composition of 15% solvent A and 85% solvent B at 10 minutes. At 12.5 minutes, the flow rate is increased to 0.5 mL/min, while maintaining a constant mobile phase composition of 15% solvent A and 85% solvent B for a period of 2.5 minutes. Finally, at 15 minutes, the flow rate is reduced back to 0.25 mL/min. To ensure system stability, the final condition is maintained for 1 minute prior to subsequent analyses. |
| Flow Rate: | 0.250 – 0.400 mL/min |
| Solvent A: | 95% acetonitrile/5%; water 20mM ammonium acetate; 20mM ammonium hydroxide (pH 9.0) |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004820 |
| Analysis ID: | AN005082 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | API |
| MS Comments: | Targeted metabolite profiling was performed using MS multiple reaction-monitoring transitions. Multiple Reaction Monitoring (MRM) Q1/Q3 peak integration of the raw data files (Analyst software, v.1.6.2; ABSciex) was performed using software MultiQuant 3.0 (ABSciex). |
| Ion Mode: | POSITIVE |
| MS ID: | MS004821 |
| Analysis ID: | AN005083 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | API |
| MS Comments: | Targeted metabolite profiling was performed using MS multiple reaction-monitoring transitions. Multiple Reaction Monitoring (MRM) Q1/Q3 peak integration of the raw data files (Analyst software, v.1.6.2; ABSciex) was performed using software MultiQuant 3.0 (ABSciex). |
| Ion Mode: | NEGATIVE |
