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MB Sample ID: SA186034
| Local Sample ID: | 5004186 |
| Subject ID: | SU002072 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002072 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 5004186 | SA186034 | FL023011 | 12 | Age |
| 5004186 | SA186034 | FL023011 | 1 | sex |
| 5004186 | SA186034 | FL023011 | CTR | Case |
Collection:
| Collection ID: | CO002065 |
| Collection Summary: | The DIABIMMUNE study recruited 832 families in Finland (Espoo), Estonia (Tartu), and Russia (Petrozavodsk) with infants carrying HLA alleles that conferred risk for autoimmunity. The subjects involved in the current study were chosen from the subset (n = 74) of international DIABIMMUNE study children who progressed to at least a single AAb (P1Ab, n = 23), who progressed to multiple islet AAb (P2Ab, n = 13), and controls (CTRs, n = 38), i.e. the children who remained islet AAb- negative during the follow-up in a longitudinal series of samples collected at 3, 6, 12, 18, 24 and 36 months from each child (Kostic et al. 2015). The study groups were matched for HLA-associated diabetes risk, sex, country and period of birth. This study was conducted according to the guidelines in the Declaration of Helsinki. The Ethics and Research Committee of the participating Universities and Hospitals approved the study protocol. All families provided written informed consent prior to sample collection. |
| Sample Type: | Blood (serum) |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002084 |
| Treatment Summary: | The bile acids were measured in serum and fecal sample as described previously (Jäntti et al., 2014; Salihović et al., 2020). |
Sample Preparation:
| Sampleprep ID: | SP002078 |
| Sampleprep Summary: | Briefly, 20 μL of serum (prepared by adding 1:20 (m/v) ultrapure water to 50 mg of feces) was filtered through a Ostro Protein Precipitation and Phospholipid Removal 96-well plate (Waters Corporation, Milford, USA), using 100 μL of cold methanol contemning the internal standard mixtures (LCA-d4, TCA-d4, GUDCA-d4, GCA-d4, CA-d4, UDCA-d4, GCDCA-d4, CDCA-d4, DCA-d4, GLCA-d4). The eluent was collected and evaporated to dryness and the residue was re-suspended in 20 μL of a 40:60 MeOH: H2O v/v mixture. The analyses were performed on an ACQUITY HSS T3 (2.1×100 mm, 1.8 μm) column, Waters (Milford), coupled to a triple quadrupole mass spectrometer (Waters Corporation, Milford, USA) with an atmospheric electrospray interface operating in negative ion mode. Separation was performed using gradient elution with 0.1 % formic acid in water (v/v) (A) and 0.1 % formic acid in acetonitrile:methanol (3:1, v/v) (B) at a flow rate of 0.5 mL/ min. Gradient program was 0 min 15 % B, 0-1 min; 30 % B, 1- 16 min; 16-18 min; 70 % B, 18–23 min 100 % B, and equilibrium time between runs was 7 min. The injection volume was 5 μL and the column was kept at 35 °C. An external calibration with nine calibration points (0.0025–600 ng/mL) was carried out for use in quantitation. |
Combined analysis:
| Analysis ID | AN003248 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Waters |
| Column | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | ABI Sciex 5500 QTrap |
| Ion Mode | NEGATIVE |
| Units | ng/ml |
Chromatography:
| Chromatography ID: | CH002393 |
| Chromatography Summary: | The analyses were performed on an ACQUITY HSS T3 (2.1×100 mm, 1.8 μm) column, Waters (Milford), coupled to a triple quadrupole mass spectrometer (Waters Corporation, Milford, USA) with an atmospheric electrospray interface operating in negative ion mode. Separation was performed using gradient elution with 0.1 % formic acid in water (v/v) (A) and 0.1 % formic acid in acetonitrile:methanol (3:1, v/v) (B) at a flow rate of 0.5 mL/ min. Gradient program was 0 min 15 % B, 0-1 min; 30 % B, 1- 16 min; 16-18 min; 70 % B, 18–23 min 100 % B, and equilibrium time between runs was 7 min. The injection volume was 5 μL and the column was kept at 35 °C. |
| Instrument Name: | Waters |
| Column Name: | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| Column Temperature: | 35 |
| Flow Gradient: | 0 min 15 % B, 0-1 min; 30 % B, 1- 16 min; 16-18 min; 70 % B, 18–23 min 100 % B, and equilibrium time between runs was 7 min. |
| Flow Rate: | 0.5 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 75% acetonitrile/25% methanol; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003021 |
| Analysis ID: | AN003248 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | The bile acids were measured in serum and fecal sample as described based on . 20 μL of serum, using the same internal standard mixtures, was filtered through a frit filter plate (96-Well Protein Precipitation Filter Plate, Sigma Aldrich), and the effluent was collected and evaporated to dryness and the residue was dissolved in 20 μL of a 40:60 MeOH: H2O v/v mixture containing the same injection standards. Analyses were performed on an ACQUITY UPLC system coupled to a triple quadrupole mass spectrometer (Waters Corporation, Milford, USA) with an atmospheric electrospray interface operating in negative ion mode. An external calibration with six calibration points (0.5–600 ng/mL), including a solvent blank, was carried out for use in quantitation. |
| Ion Mode: | NEGATIVE |
