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MB Sample ID: SA327115
Local Sample ID: | DS1-27-10 |
Subject ID: | SU003126 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Female |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU003126 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Female |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
DS1-27-10 | SA327115 | FL038855 | Pre-diet | factor |
Collection:
Collection ID: | CO003119 |
Collection Summary: | Blood samples were collected from consented study subjects and plasma isolated according to standard clinical laboratory protocols. |
Sample Type: | Blood (plasma) |
Treatment:
Treatment ID: | TR003135 |
Treatment Summary: | This study was a clinical trial (NCT02653092) and was approved by the Colorado Multiple Institutional Review Board (COMIRB). All participants provided informed consent. Women aged 18-38 with menstrual cycles 25-35 days in length and no use of reproductive hormones within 3 months of enrollment were eligible for the study. Additional eligibility criteria included: a) no history of chronic disease affecting hormone production, metabolism, or clearance and no use of medications known to interact with reproductive hormones or insulin metabolism; b) normal screening prolactin and TSH; c) normal hemoglobin A1c; d) hemoglobin > 11 g/dl at screening; e) use of reliable non-hormonal contraception throughout the study period. Because the goal was to investigate the effects of a high-fat diet, women with a baseline dietary assessment indicative of >40% calories from fat were excluded, as were women who were unable to comply with the protocol, which involved eating only foods provided by the Colorado Clinical and Translational Sciences Institute’s (CCTSI) Nutrition Services. Because of the restrictions on their ability to consume animal or dairy fats, vegans and lactose intolerant individuals were excluded. Cycle 1 was a baseline cycle; Cycle 2 entailed consumption of the high-fat diet which was continued until frequent blood sampling was completed in the early follicular phase of Cycle 3. Cycle 3 was the immediate post-high-fat-diet cycle and Cycle 4 was a recovery cycle. Participants underwent baseline and end-of-diet assessments of gonadotropin dynamics. A 6-hour frequent blood sampling study was performed to determine endogenous LH and FSH secretion (Time 0- 341 240 minutes) and concluded with a physiologic bolus of 75 ng/kg GnRH to assess pituitary 342 responsiveness (Time 240-360 minutes). Frequent sampling studies for gonadotropin determination were 343 all performed in the early follicular phase of the menstrual cycle (cycle days 2-5). Participants were also 344 seen weekly during the high-fat diet cycle and had a blood draw to assess compliance. 345 346 High-fat diet protocol. Participants completed a food preference screening survey and a 3-day diet diary, 347 which estimated their daily caloric intake and approximate proportion of daily calories derived from fat. 348 Participants were provided with a customized eucaloric high-fat diet comprised of approximately 48% 349 calories from fat, 33% calories from carbohydrates and 19% calories from protein. Food was portioned 350 into individual meals with snacks and boxed into 3-4 days’ worth at a time. Participants picked up food 351 directly from the Clinical and Translational Research Center; when this was not possible, study staff 352 delivered food to the participant. Urinary ketones were measured at each frequent blood sampling 353 session (Siemens Healthineers, Malvern, PA). During administration of the high-fat diet, blood samples were drawn weekly for 364 metabolomic analysis to determine compliance with the protocol. |
Sample Preparation:
Sampleprep ID: | SP003132 |
Sampleprep Summary: | Plasma for metabolomics was obtained after an overnight fast and 20 μl was extracted in 980 μl of cold methanol:acetonitrile:water (5:3:2, v/v/v). After vortexing at 4°C for 30 min, extracts were separated from the protein pellet by centrifugation for 10 min at 18,000 g at 4°C and stored at −80°C until analysis. |
Processing Storage Conditions: | 4℃ |
Extract Storage: | -80℃ |
Combined analysis:
Analysis ID | AN004944 | AN004945 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Reversed phase | Reversed phase |
Chromatography system | Thermo Vanquish | Thermo Vanquish |
Column | Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) | Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) |
MS Type | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap |
MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
Ion Mode | NEGATIVE | POSITIVE |
Units | peak area | peak area |
Chromatography:
Chromatography ID: | CH003732 |
Chromatography Summary: | Negative C18 |
Instrument Name: | Thermo Vanquish |
Column Name: | Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) |
Column Temperature: | 45 |
Flow Gradient: | 0-0.5 min 0% B, 0.5-1.1 min 0-100% B, 1.1-2.75 min hold at 100% B, 2.75-3 min 100-0% B, 3-5 min hold at 0% B |
Flow Rate: | 450 uL/min |
Sample Injection: | 6 uL |
Solvent A: | 95% water 5% acetonitrile 1 mM ammonium acetate |
Solvent B: | 95% acetonitrile 5% water 1 mM ammonium acetate |
Chromatography Type: | Reversed phase |
Chromatography ID: | CH003733 |
Chromatography Summary: | Positive C18 |
Instrument Name: | Thermo Vanquish |
Column Name: | Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) |
Column Temperature: | 45 |
Flow Gradient: | 0-0.5 min 5% B, 0.5-1.1 min 5-95% B, 1.1-2.75 min hold at 95% B, 2.75-3 min 95-5% B, 3-5 min hold at 5% B |
Flow Rate: | 450 uL/min |
Sample Injection: | 6 uL |
Solvent A: | 100% water; 0.1% formic acid |
Solvent B: | 100%acetonitrile; 0.1% formic acid |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS004687 |
Analysis ID: | AN004944 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Resolution 70,000, scan range 65-900 m/z, maximum injection time 200 ms, microscans 2, automatic gain control (AGC) 3 x 10^6 ions, source voltage 4.0 kV, capillary temperature 320 C, and sheath gas 45, auxiliary gas 15, and sweep gas 0 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were annotated and integrated using Maven in conjunction with the KEGG database. |
Ion Mode: | NEGATIVE |
MS ID: | MS004688 |
Analysis ID: | AN004945 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Resolution 70,000, scan range 65-900 m/z, maximum injection time 200 ms, microscans 2, automatic gain control (AGC) 3 x 10^6 ions, source voltage 4.0 kV, capillary temperature 320 C, and sheath gas 45, auxiliary gas 15, and sweep gas 0 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were annotated and integrated using Maven in conjunction with the KEGG database. |
Ion Mode: | POSITIVE |