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MB Sample ID: SA085904
Local Sample ID: | 309-02 P |
Subject ID: | SU001282 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Male and female |
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Subject:
Subject ID: | SU001282 |
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 |
---|---|---|---|---|
309-02 P | SA085904 | FL012840 | Placebo | Treatment |
Collection:
Collection ID: | CO001276 |
Collection Summary: | Human plasma was acquired from a previously performed clinical trial (Cypess et al., 2015) registered with ClinicalTrials.gov (NCT01783470) and had the FDA Investigational New Drug (IND) registration number 116246. It was approved by the Human Studies Institutional Review Boards of Beth Israel Deaconess Medical Center (BIDMC) and Joslin Diabetes Center (JDC). Blood samples were collected 180 minutes after oral dosing. 180 minutes is the corresponding T max (time after administration of a drug when the maximum plasma concentration is reached) found for mirabegron in men (Baskin et al., Diabetes 2018). |
Sample Type: | Blood (plasma) |
Treatment:
Treatment ID: | TR001297 |
Treatment Summary: | Healthy volunteers were recruited through electronic advertisements and provided written informed consent. The subjects were given a single oral dose of mirabegron, 200 mg. |
Sample Preparation:
Sampleprep ID: | SP001290 |
Sampleprep Summary: | Aliquots of 100 µL plasma were taken, depending on the experiment. A mixture of deuterium-labeled internal standards was added to each aliquot, followed by 3x volume of sample of cold methanol (MeOH). Samples were vortexed for 5 minutes and stored at 31 −20 °C overnight. Cold samples were centrifuged at 14,000g for 10 minutes, and the supernatant was then transferred to a new tube and 3 mL of acidified H 2 O (pH 3.5) was added to each sample prior to C18 SPE. The methyl formate fractions were collected, dried under nitrogen, and reconstituted in 50 µL MeOH:H 2 O (1:1, by vol). Samples were transferred to 0.5 mL tubes and centrifuged at 20,000g at 4 °C for 10 minutes. (35ul) of supernatant was transferred to LC–MS/MS vials for analysis using the BERG LCMS/MS mediator lipidomics platform. |
Combined analysis:
Analysis ID | AN002026 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Ekspert MicroLC 200 system |
Column | Phenomenex Synergi Fusion-RP capillary C18 (150 × 0.5 mm,4um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | ABI Sciex 5600+ TripleTOF |
Ion Mode | NEGATIVE |
Units | Peak Area |
Chromatography:
Chromatography ID: | CH001467 |
Instrument Name: | Ekspert MicroLC 200 system |
Column Name: | Phenomenex Synergi Fusion-RP capillary C18 (150 × 0.5 mm,4um) |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS001879 |
Analysis ID: | AN002026 |
Instrument Name: | ABI Sciex 5600+ TripleTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | MS analysis was performed on a SCIEX TripleTOF® 5600+ system using the HR-MRM strategy consisting of a TOF MS experiment looped with multiple MS/MS experiments. MS spectra were acquired in high-resolution mode (>30,000) using a 100-ms accumulation time per spectrum. Fullscan MS/MS was acquired in high sensitivity mode, with an accumulation time optimized per cycle. Collision energy was set using rolling collision energy with a spread of 15V. The identity of a component was confirmed using PeakView® software (SCIEX), and quantification was performed using MultiQuant™ software (SCIEX). |
Ion Mode: | NEGATIVE |