Summary of Study ST000600

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench,, where it has been assigned Project ID PR000437. The data can be accessed directly via it's Project DOI: 10.21228/M8K31K This work is supported by NIH grant, U2C- DK119886.


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Study IDST000600
Study TitleNEFA Profile Response to Triphenyl Phosphate Exposure
Study SummaryThis study aims to identify changes in non-esterified fatty acid (NEFAs) in the plasma with triphenyl phosphate (TPP) exposure. UC Davis type 2 diabetes mellitus (UCD-T2DM) rats were treated with TPP or not treated. Each group was analyzed for non-esterified fatty acid (NEFA) changes to investigate alterations in NEFAs due to TPP exposure. Targeted analysis of NEFA in rat plasma samples was performed by the Newman lab.
University of California, Davis
DepartmentUSDA Western Human Nutrition Research Center
Last NameNewman
First NameJohn
Address430 W. Health Sciences Dr., Davis, CA 95616
Submit Date2017-04-27
Study CommentsThe samples included a high degree of hemolysis exhibited in the plasma. One sample was lost during processing (Group E- Subject 78). Two samples were outliers for multiple analytes and were not included in the final data (E-117 & T-28). Of the samples reported in this data set, there were no missing values.
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailGC-MS
Release Date2017-07-10
Release Version1
John Newman John Newman application/zip

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Project ID:PR000437
Project DOI:doi: 10.21228/M8K31K
Project Title:NEFA Profile Response to Triphenyl Phosphate Exposure
Project Summary:This study aims to identify changes in non-esterified fatty acid (NEFAs) in the plasma with triphenyl phosphate (TPP) exposure. UC Davis type 2 diabetes mellitus (UCD-T2DM) rats were treated with TPP or not treated. Each group was analyzed for non-esterified fatty acid (NEFA) changes to investigate alterations in NEFAs due to TPP exposure. Targeted analysis of NEFA in rat plasma samples was performed by the Newman lab.
Institute:University of California, Davis
Department:Department of Environmental Toxicology
Last Name:La Merrill
First Name:Michele
Address:1 Shields Ave., Davis, CA 95616
Phone:(530) 754-7254


Subject ID:SU000623
Subject Type:Animal
Subject Species:Rattus norvegicus
Taxonomy ID:10116
Species Group:Mammal


Subject type: Animal; Subject species: Rattus norvegicus (Factor headings shown in green)

mb_sample_id local_sample_id Treatment
SA033113LM-24E (Vehicle control)
SA033114LM-22E (Vehicle control)
SA033115LM-14E (Vehicle control)
SA033116LM-16E (Vehicle control)
SA033117LM-15E (Vehicle control)
SA033118LM-01E (Vehicle control)
SA033119LM-10E (Vehicle control)
SA033120LM-31E (Vehicle control)
SA033121LM-13E (Vehicle control)
SA033122LM-12E (Vehicle control)
SA033123LM-09E (Vehicle control)
SA033124LM-06E (Vehicle control)
SA033125LM-08E (Vehicle control)
SA033126LM-32E (Vehicle control)
SA033127LM-19T (Treated with TPhP)
SA033128LM-03T (Treated with TPhP)
SA033129LM-25T (Treated with TPhP)
SA033130LM-07T (Treated with TPhP)
SA033131LM-21T (Treated with TPhP)
SA033132LM-29T (Treated with TPhP)
SA033133LM-11T (Treated with TPhP)
SA033134LM-20T (Treated with TPhP)
SA033135LM-27T (Treated with TPhP)
SA033136LM-05T (Treated with TPhP)
SA033137LM-02T (Treated with TPhP)
SA033138LM-28T (Treated with TPhP)
SA033139LM-17T (Treated with TPhP)
SA033140LM-26T (Treated with TPhP)
SA033141LM-30T (Treated with TPhP)
Showing results 1 to 29 of 29


Collection ID:CO000617
Collection Summary:Prior to sacrifice, rats were fasted between 8 and 12 h,their body weights recorded, and blood was collected from the tail as above prior to euthanasia. Rats were anesthetized with sodium pentobarbital and euthanized with a 1 mL/kg intracardiac injection of saturated potassium chloride. Once cardiac movement had stopped for 30 s the rat was decapitated and the hypothalamus, liver, pancreas, heart, mesenteric adipose tissue, quadriceps, kidney, gonadal adipose tissue, inguinal adipose tissue, and brown adipose tissue were collected. All tissues were removed in the order listed above, wet weighed, and snap frozen in liquid nitrogen
Collection Protocol Filename:Green_et_al_2017-TPP_Exposure_Accelerat_T2DM_Rats.pdf
Sample Type:Blood
Blood Serum Or Plasma:Plasma


Treatment ID:TR000637
Treatment Summary:Adult non-pregnant female UCD-T2DM rats (n = 16; 3 months old) were paired with males (n = 10; 3–4 months old) for a 24 h period at which point males were removed. This was defined as gestational day zero (G0) if a sperm plug was observed or if the female rats gained at least 30 g of body weight over the next 7 days. The day of birth was designated postnatal day zero (P0). Pregnant dams were randomly assigned to an exposure group (n = 8 per group), and received daily oral TPhP or ethanol vehicle exposure from G8 through weaning (P21) as described in Section 2.2 below. Gestational length and litter size were recorded on P0 and the sex of pups was determined and recorded on P4. Body weights of all pups in each litter were obtained periodically from P4–21. On P4 the litters were culled to 8 pups ensuring up to 4males and 2 females in each litter by random selection (Fig. 1A & B). This was done to ensure consistent exposure of pups between litters [13,23]. The time ittakes to develop T2DM is accelerated among UCD-T2DM rats with higher body weights on P21. Hence at weaning the largest pups were housed in same sex littermate groups of two females and up to four males as available (Fig. 1A & B). Urine was collected from the dams using an adapted plastic wrap method outlined by Kurien [24], 60 mins after final dose. Dams were placed in clean cages without bedding for at least 20 min then using a pipette up to 500 L of urine was collected in ethanol rinsed glass vials and placed on ice. At weaning all dams and remaining weanlings were sacrificed (90–330 min post-exposure) by CO2 asphyxiation and rapid decapitation. Two male rats weighing between 350–400 g on P61, from the TPhP group and the vehicle group were weight-matched across treatments for the diabetes study to eliminate confounding effects of body mass on the association between TPhP and T2DM onset (Fig. 1B). This weight range was selected because male UCD-T2DM rats that are between 350 and 400 g at 8 weeks of age develop T2DM at approximately 23 weeks of age [18]. Weight-matched rats were followed until 26 weeks or until they developed T2DM, which was defined as two consecutive weekly non-fasting glucose measurements of ≥200 mg/dL [18] in accordance with the American Diabetes Association (ADA) guideline of diagnosing diabetes with a random plasma glucose of 200 mg/dL or higher [19]. The remaining rats were not weight-matched and followed for the 3.5 month obesity study (Fig. 1A).
Treatment Protocol Filename:Green_et_al_2017-TPP_Exposure_Accelerat_T2DM_Rats.pdf

Sample Preparation:

Sampleprep ID:SP000630
Sampleprep Summary:Plasma non-esterified fatty acids (NEFAs) were isolated as previously described by Smedes (1). and Gladine et al. (2). Specifically, plasma aliquots (100 mL) were enriched with 5 mL 0.2 mg/ml butylated hydroxytoluene/EDTA in 1:1 methanol:- water, and a suite of extraction surrogates, which included deuterated-tri-palmitoyl glycerol (d31-16:0-TG; CDN Isotopes, Pointe-Claire, Quebec, Canada), deuterated distearoylphosphotidylcholine (d35-18:0-PC; Avanti Polar Lipids, Alabaster, Alabama), dodeca-(9E)-enoyl cholesterylesters (22:1n9-CE; NuChek Prep, Elysian MN) and dodecatrienoic acid (22:3n3; NuChek Prep). Lipids were then extracted with 10:8:11 cylcohexane: 2- propanol:ammonium acetate. Briefly, enriched samples were mixed with cyclopropane/2-propanol, phases were split with ammonium acetate, the organic phase was isolated and the aqueous phase was re-extracted with cyclohexane. The combined organic total lipid extract was reduced to dryness and reconstituted in 200 µL of 1:1 methanol/toluene and the total lipid extract was used to quantify plasma fatty acids as methyl esters by gas chromatography-mass spectrometry (GC-MS). It was derivitized by adding 45 μL 2M (trimethylsilyl) diazomethane in hexanes and spiked with 15:1n5 free acid to track methylation efficiency. Next, it was brought to a final volume of 200 mL with 90:10 methanol/toluene (v/v) and left at room temperature for 30 min, before being brought to dryness. The remaining fatty acid methyl esters (FAMEs) were re-constituted in 300 mL Hexane plus 10 uL of 44 mM tricosanoate methyl ester (23:0; NuChek Prep), vortexed, and 100 uL was transferred to a GC-MS Vial for analysis.
Sampleprep Protocol Filename:NEFA_Plasma_Newman_Data_Report.docx

Combined analysis:

Analysis ID AN000919
Analysis type MS
Chromatography type GC
Chromatography system Agilent 6890N
Column Agilent DB-225 (30m x 0.25mm x 0.5um)
MS Type EI
MS instrument type GC-TOF
MS instrument name Agilent
Units Concentration (µM)


Chromatography ID:CH000655
Instrument Name:Agilent 6890N
Column Name:Agilent DB-225 (30m x 0.25mm x 0.5um)
Sample Injection:1 µl
Chromatography Type:GC


MS ID:MS000818
Analysis ID:AN000919
Instrument Name:Agilent
Instrument Type:GC-TOF
MS Type:EI