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MB Sample ID: SA192296
| Local Sample ID: | NHF-6 |
| Subject ID: | SU002126 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002126 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| NHF-6 | SA192296 | FL023603 | NA | Gender |
Collection:
| Collection ID: | CO002119 |
| Collection Summary: | Serum samples were collected from CVD patients at the Safdarjung Hospital, New Delhi. In all assessments, all individuals were asked to provide complete information regarding personal health history, self-rated health, and family history of diabetes, tobacco use, and physical activity. Ethical approval was obtained at Safdarjung Hospital, New Delhi for sample collection. |
| Sample Type: | Blood (serum) |
Treatment:
| Treatment ID: | TR002138 |
| Treatment Summary: | NA |
Sample Preparation:
| Sampleprep ID: | SP002132 |
| Sampleprep Summary: | Serum samples (100µl) should be properly thawed and spin at 5K rpm for one minute, added with 100 % MeOH (300µl, in1:3 ratio) and vortex well (30s), incubated on ice for 10 minutes and then centrifuged at 10K rpm for 10 minutes at 4℃. Equally, transfer the supernatant into two different well-labeled tubes and then speed vacuum dry and stored at -80℃. One set of the tubes is used for the analysis of polar metabolites and the other one is used for the non-polar metabolites. For the reconstitution of polar metabolites, 50% ACN (50 µl) is added into the vacuum dried sample and for nonpolar 15% of MeOH (50 µl) is used. After the addition of reconstitutes, vortex well for 10 min and centrifuged at 10K rpm for 10 minutes at 4℃ and transfer the supernatant (approx. 40-45 µl) into MS vials. Remove 10µl form every sample containing MS vials and pooled together into a 15 ml tube to use as QC. Then, the samples were stored at -80℃ for further analysis. |
Combined analysis:
| Analysis ID | AN003325 | AN003326 | AN003327 | AN003328 |
|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS |
| Chromatography type | HILIC | HILIC | Reversed phase | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
| Column | Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um) | Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um) | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI | ESI | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap | Orbitrap | Orbitrap |
| MS instrument name | Thermo Fusion Orbitrap | Thermo Fusion Orbitrap | Thermo Fusion Orbitrap | Thermo Fusion Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE | POSITIVE | NEGATIVE |
| Units | peak height | peak height | peak height | peak height |
Chromatography:
| Chromatography ID: | CH002464 |
| Chromatography Summary: | For hydrophilic interaction (HILIC) chromatography, 5 µl of the prepared samples were injected for analysis. An ultimate 3000 UPLC XBridge BHE Amide column (2.1 x 100 mm, 2.5 µm; Waters Corporation) was used and maintained at 35℃ for separation. The mobile phase was delivered at 0.35mL/min. The mobile phase A consisted of 20mM ammonium acetate in the water of pH 9.0 and mobile phase B was 100% acetonitrile. The elution gradient is used as follows: 0 min-85% B, 2 min-10% B, 14 min- 10% B, 14.1 min- 85% B, 16 min- 85%B. |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um) |
| Column Temperature: | 35 |
| Flow Gradient: | 0 min-85% B, 2 min-10% B, 14 min- 10% B, 14.1 min- 85% B, 16 min- 85%B. |
| Flow Rate: | 0.35mL/min |
| Solvent A: | 100% water; 20 mM ammonium acetate, pH 9.0 |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH002465 |
| Chromatography Summary: | For reversed-phase (RP) chromatographic analyses, 5 µL of prepared samples were injected. The separation was performed using a UPLC ultimate 3000 using the HSS T3 column (2.1x 100mm, 1.7 µm; Waters Corporation) at 40℃. The mobile phase was delivered at 300µL/min and consisted of eluent A (water with 0.1%formic acid) and eluent B (acetonitrile with 0.1% formic acid), delivered in a gradient profile: 0min-1% B, 1min-15%B, 4min-35% B, 7min-95% B, 9 min- 95% B, 10min-1% B, 14 min- 1% B. |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
| Flow Gradient: | 0min-1% B, 1min-15%B, 4min-35% B, 7min-95% B, 9 min- 95% B, 10min-1% B, 14 min- 1% B |
| Flow Rate: | 300µL/min |
| Solvent A: | 100% water; 0.1%formic acid |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003095 |
| Analysis ID: | AN003325 |
| Instrument Name: | Thermo Fusion Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The electrospray ionization (ESI) source was operated for both positive mode (+) and negative (-) ionization at 120,000 resolution in MS1 mode and 30000 resolution in data-dependent MS2 scan mode. The spray voltage used for these positive and negative modes is 4000 and 35000 volts respectively. Sheath gas and auxiliary gas were set to 42 and 11 respectively. The mass scan range was 60-900 m/z, ACG (Automatic control gain) target at 100000 ions and maximum injection time was 50 ms for MS and ACG target was 20000 ions and maximum injection time 60 ms for MSMS was used. Data were processed using the IDSL.IPA software (https://cran.r-project.org/web/packages/IDSL.IPA/index.html) |
| Ion Mode: | POSITIVE |
| MS ID: | MS003096 |
| Analysis ID: | AN003326 |
| Instrument Name: | Thermo Fusion Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The electrospray ionization (ESI) source was operated for both positive mode (+) and negative (-) ionization at 120,000 resolution in MS1 mode and 30000 resolution in data-dependent MS2 scan mode. The spray voltage used for these positive and negative modes is 4000 and 35000 volts respectively. Sheath gas and auxiliary gas were set to 42 and 11 respectively. The mass scan range was 60-900 m/z, ACG (Automatic control gain) target at 100000 ions and maximum injection time was 50 ms for MS and ACG target was 20000 ions and maximum injection time 60 ms for MSMS was used. Data were processed using the IDSL.IPA software (https://cran.r-project.org/web/packages/IDSL.IPA/index.html) |
| Ion Mode: | NEGATIVE |
| MS ID: | MS003097 |
| Analysis ID: | AN003327 |
| Instrument Name: | Thermo Fusion Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The electrospray ionization (ESI) source was operated for both positive mode (+) and negative (-) ionization at 120,000 resolution in MS1 mode and 30000 resolution in data-dependent MS2 scan mode. The spray voltage used for these positive and negative modes is 4000 and 35000 volts respectively. Sheath gas and auxiliary gas were set to 42 and 11 respectively. The mass scan range was 60-900 m/z, ACG (Automatic control gain) target at 100000 ions and maximum injection time was 50 ms for MS and ACG target was 20000 ions and maximum injection time 60 ms for MSMS was used. Data were processed using the IDSL.IPA software (https://cran.r-project.org/web/packages/IDSL.IPA/index.html) |
| Ion Mode: | POSITIVE |
| MS ID: | MS003098 |
| Analysis ID: | AN003328 |
| Instrument Name: | Thermo Fusion Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The electrospray ionization (ESI) source was operated for both positive mode (+) and negative (-) ionization at 120,000 resolution in MS1 mode and 30000 resolution in data-dependent MS2 scan mode. The spray voltage used for these positive and negative modes is 4000 and 35000 volts respectively. Sheath gas and auxiliary gas were set to 42 and 11 respectively. The mass scan range was 60-900 m/z, ACG (Automatic control gain) target at 100000 ions and maximum injection time was 50 ms for MS and ACG target was 20000 ions and maximum injection time 60 ms for MSMS was used. Data were processed using the IDSL.IPA software (https://cran.r-project.org/web/packages/IDSL.IPA/index.html) |
| Ion Mode: | NEGATIVE |
