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MB Sample ID: SA168889
Local Sample ID: | C62B |
Subject ID: | SU001891 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001891 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
C62B | SA168889 | FL020086 | CONTROL | Factor1 |
Collection:
Collection ID: | CO001884 |
Collection Summary: | On the day of the interview subjects were appointed for blood sample analysis within the next week at the primary care centre, where the interview took place, and a 30 ml blood sample were obtained at baseline. Fasting venous blood samples were drawn and immediately processed and were divided into 0.5 ml aliquots of plasma, serum, concentrated red blood cells and buffy coat, and stored in liquid nitrogen tanks at –190ºC until analysis. |
Sample Type: | Blood (plasma) |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR001904 |
Treatment Summary: | Briefly, for GC-MS analysis, protein precipitation was achieved by mixing 1 volume of plasma with 3 volumes of cold (-20°C) acetonitrile, followed by methoximation with O-methoxyamine hydrochloride (15 mg/mL) in pyridine, and silylation with BSTFA:TMSC (99:1). Finally, 20 ppm of tricosane in heptane was added as internal standard (IS). For CE-MS analysis, 100 µL of plasma was mixed with 100 µL of 0.2 M formic acid containing 5% acetonitrile and 0.4 mM methionine sulfone as IS. The sample was transferred to a centrifree ultracentrifugation device (Millipore Ireland Ltd.,Carrigtohill, Ireland) with a 30 kDa protein cutoff for deproteinization through centrifugation (2000 g, 4 °C, 70 min). For LC-MS analysis, 100 µL of plasma was mixed with 300 µL of a cold mixture (-20°C) of methanol:ethanol (1:1, v/v) for deproteinization. Samples were centrifuged (13,000 g, 4°C, 20 min). After centrifugation, 100 µL of the supernatant was directly injected into the system. |
Sample Preparation:
Sampleprep ID: | SP001897 |
Sampleprep Summary: | for GC-MS analysis, protein precipitation was achieved by mixing 1 volume of plasma with 3 volumes of cold (-20°C) acetonitrile, followed by methoximation with O-methoxyamine hydrochloride (15 mg/mL) in pyridine, and silylation with BSTFA:TMSC (99:1). Finally, 20 ppm of tricosane in heptane was added as internal standard (IS). For CE-MS analysis, 100 µL of plasma was mixed with 100 µL of 0.2 M formic acid containing 5% acetonitrile and 0.4 mM methionine sulfone as IS. The sample was transferred to a centrifree ultracentrifugation device (Millipore Ireland Ltd.,Carrigtohill, Ireland) with a 30 kDa protein cutoff for deproteinization through centrifugation (2000 g, 4 °C, 70 min). For LC-MS analysis, 100 µL of plasma was mixed with 300 µL of a cold mixture (-20°C) of methanol:ethanol (1:1, v/v) for deproteinization. Samples were centrifuged (13,000 g, 4°C, 20 min). After centrifugation, 100 µL of the supernatant was directly injected into the system. |
Combined analysis:
Analysis ID | AN002940 | AN002941 | AN002942 | AN002943 |
---|---|---|---|---|
Analysis type | MS | MS | MS | MS |
Chromatography type | Reversed phase | Reversed phase | GC | Normal phase |
Chromatography system | Agilent 1200 | Agilent 1200 | Agilent 7890A | Agilent 7100 CE |
Column | Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) | Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) | Agilent DB5-MS (30m x 0.25mm, 0.25um) | Agilent Technologies fused silica capillary 96cm x 50μm |
MS Type | ESI | ESI | EI | ESI |
MS instrument type | QTOF | QTOF | Single quadrupole | TOF |
MS instrument name | Agilent 6520 QTOF | Agilent 6520 QTOF | Agilent 5975C | Agilent 6224 TOF |
Ion Mode | POSITIVE | NEGATIVE | UNSPECIFIED | POSITIVE |
Units | AREA | AREA | AREA |
Chromatography:
Chromatography ID: | CH002177 |
Chromatography Summary: | The analysis of the samples was accomplished using an UHPLC system (1200 Infinity system, Agilent Technologies, Waldbronn, Germany), coupled to a 6520 QTOF MS (Agilent Technologies) with an ESI ion source. The sample injection volume was set up to 10 μL. The separation was achieved using a Discovery® HS C18 15cm x 2.1 mm, 3 µm (Supelco analytical) reverse phase column at thermostated 40 °C. |
Instrument Name: | Agilent 1200 |
Column Name: | Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) |
Chromatography Type: | Reversed phase |
Chromatography ID: | CH002178 |
Chromatography Summary: | An Agilent GC system (7890A) coupled to a- 5975C mass spectrometer (Agilent Technologies) was used to perform metabolite fingerprinting of plasma samples. Briefly, 2 μL of derivatized samples were automatically injected in split mode (ratio 1:10) through a split liner of ultra-inert deactivated glass wool from Agilent. The separation of the compounds was achieved using a pre-column (10 m J&W integrated with Agilent 122‐5532G) combined with a GC DB5-MS column (length, 30 m; internal diameter, 0.25 mm; and 0.25 μm film of 95% of dimethyl/5% diphenylpolysiloxane). |
Instrument Name: | Agilent 7890A |
Column Name: | Agilent DB5-MS (30m x 0.25mm, 0.25um) |
Chromatography Type: | GC |
Chromatography ID: | CH002179 |
Chromatography Summary: | The analysis was performed using a 7100 capillary electrophoresis (Agilent Technologies) coupled to a TOF MS 6224 mass spectrometer (Agilent Technologies), equipped with an ESI ion source. For the separation of metabolites, an Agilent Technologies fused silica capillary (total length, 96 cm; internal diameter, 50 μm) was used, working in normal polarity. Before each analysis, the capillary was washed for 5 min (950 mbar) with background electrolyte (BGE) (0.8 M formic acid solution in 10% methanol (v/v)). The sample injection was performed during 50 s at 50 mbar and, in order to improve the reproducibility of the analysis, the BGE was injected for 20 s at 100 mbar after the injection of each sample. |
Instrument Name: | Agilent 7100 CE |
Column Name: | Agilent Technologies fused silica capillary 96cm x 50μm |
Chromatography Type: | Normal phase |
MS:
MS ID: | MS002730 |
Analysis ID: | AN002940 |
Instrument Name: | Agilent 6520 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | Agilent Technologies MassHunter WorkStation Data Acquisition B.08.00 software; Agilent Technologies MassHunter Profinder B.08.00 SP1 software (Waldbronn, Germany) |
Ion Mode: | POSITIVE |
MS ID: | MS002731 |
Analysis ID: | AN002941 |
Instrument Name: | Agilent 6520 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | Agilent Technologies MassHunter WorkStation Data Acquisition B.08.00 software; Agilent Technologies MassHunter Profinder B.08.00 SP1 software (Waldbronn, Germany) |
Ion Mode: | NEGATIVE |
MS ID: | MS002732 |
Analysis ID: | AN002942 |
Instrument Name: | Agilent 5975C |
Instrument Type: | Single quadrupole |
MS Type: | EI |
MS Comments: | Agilent MSD ChemStation software (Agilent Technologies); Agilent MassHunter Qualitative B.08.00; Agilent MassHunter Unknowns Analysis Tool 9.0. |
Ion Mode: | UNSPECIFIED |
MS ID: | MS002733 |
Analysis ID: | AN002943 |
Instrument Name: | Agilent 6224 TOF |
Instrument Type: | TOF |
MS Type: | ESI |
MS Comments: | Agilent Technologies MassHunter WorkStation Data Acquisition B.08.00 software; Agilent Technologies MassHunter Profinder B.08.00 SP1 software (Waldbronn, Germany) |
Ion Mode: | POSITIVE |