Summary of Study ST002920

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 PR001814. The data can be accessed directly via it's Project DOI: 10.21228/M87D9M This work is supported by NIH grant, U2C- DK119886.


This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST002920
Study TitlePossible PPARG-independent effects of DINCH and MINCH on central carbon metabolism
Study SummaryIn the third part of the project, we investigated the PPARG-independent effects of DINCH and MINCH on the central carbon metabolism of human SGBS cells. SGBS preadipocytes were treated for 12 days with 10 µM MINCH, 10 µM DINCH, or rosiglitazone in the presence of the PPARG inhibitor GW9662. Irreversible blocking of PPARG was achieved by incubating cells with 10 µM GW9662 1 hour before treatment and maintaining a regular treatment interval of 2 days. In conclusion, although the PPARG inhibitor GW9662 greatly reduced the effects of MINCH and rosiglitazone, slightly increased lipid accumulation along with increased lactate secretion and increased concentrations of pyruvate cycle metabolites were consistently induced by MINCH treatment even after PPARG inhibition. This suggests that MINCH exerts its effects on lipid accumulation and central carbon metabolism at least in part via a PPARG-independent mechanism.
Helmholtz Centre for Environmental Research
Last NameEngelmann
First NameBeatrice
AddressPermoserstr. 15
Submit Date2023-10-08
Raw Data AvailableYes
Raw Data File Type(s)wiff
Analysis Type DetailLC-MS
Release Date2023-11-01
Release Version1
Beatrice Engelmann Beatrice Engelmann application/zip

Select appropriate tab below to view additional metadata details:


Project ID:PR001814
Project DOI:doi: 10.21228/M87D9M
Project Title:MINCH causes metabolic rewiring towards lipid accumulation and adipogenesis
Project Summary:Humans are ubiquitously exposed to plastic additives, including plasticizers. There is growing evidence that exposure to certain plasticizers is associated with the development of obesity due to their metabolism-disrupting properties. Following the restriction of the use of the phthalate plasticizer di-(2-ethylhexyl) phthalate (DEHP) due to its adverse health effects, it has been replaced by new substitutes such as the plasticizer diisononylcyclohexane-1,2-dicarboxylate (DINCH). Despite recent studies suggesting that the primary metabolite monoisononylcyclohexane-1,2-dicarboxylic acid ester (MINCH) promotes human adipocyte differentiation, the adipogenic properties of MINCH remain controversial. Because the metabolome largely reflects the molecular phenotype and is sensitive to perturbation by external factors, we used targeted metabolomics to investigate the effects of DINCH and MINCH on key metabolic pathways of adipocytes. Analysis of central carbon metabolism is particularly relevant because it provides cellular energy through the degradation of organic compounds and metabolic precursors for anabolic functions that are critical for adipocyte function, such as de novo lipogenesis. The project consists of three main studies: analysis of the effects of DINCH and MINCH on central carbon metabolism of human SGSB cells, analysis of the insulin response of DINCH- and MINCH-treated SGSB cells, and analysis of the effects of DINCH and MINCH on central carbon metabolism of human SGSB cells in the presence of the PPARG inhibitor GW9662.
Institute:Helmholtz Centre for Environmental Research
Last Name:Engelmann
First Name:Beatrice
Address:Permoserstr. 15


Subject ID:SU003033
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606


Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Treatment
SA316858Ctrl_GW_1Control GW9662 treatment
SA316859Ctrl_GW_4Control GW9662 treatment
SA316860Ctrl_GW_3Control GW9662 treatment
SA316861Ctrl_GW_2Control GW9662 treatment
SA316862Ctrl_GW_SN2Control GW9662 treatment supernatant
SA316863Ctrl_GW_SN4Control GW9662 treatment supernatant
SA316864Ctrl_GW_SN3Control GW9662 treatment supernatant
SA316865Ctrl_GW_SN1Control GW9662 treatment supernatant
SA316866DINCH_GW_4DINCH 10µM + GW9662 treatment
SA316867DINCH_GW_3DINCH 10µM + GW9662 treatment
SA316868DINCH_GW_1DINCH 10µM + GW9662 treatment
SA316869DINCH_GW_2DINCH 10µM + GW9662 treatment
SA316870DINCH_GW_SN3DINCH 10µM + GW9662 treatment supernatant
SA316871DINCH_GW_SN4DINCH 10µM + GW9662 treatment supernatant
SA316872DINCH_GW_SN2DINCH 10µM + GW9662 treatment supernatant
SA316873DINCH_GW_SN1DINCH 10µM + GW9662 treatment supernatant
SA316874MINCH_GW_4MINCH 10µM + GW9662 treatment
SA316875MINCH_GW_1MINCH 10µM + GW9662 treatment
SA316876MINCH_GW_2MINCH 10µM + GW9662 treatment
SA316877MINCH_GW_3MINCH 10µM + GW9662 treatment
SA316878MINCH_GW_SN2MINCH 10µM + GW9662 treatment supernatant
SA316879MINCH_GW_SN3MINCH 10µM + GW9662 treatment supernatant
SA316880MINCH_GW_SN4MINCH 10µM + GW9662 treatment supernatant
SA316881MINCH_GW__SN1MINCH 10µM + GW9662 treatment supernatant
SA316882Rosi_GW_4Rosi (d0-d4) + GW9662 treatment
SA316883Rosi_GW_1Rosi (d0-d4) + GW9662 treatment
SA316884Rosi_GW_3Rosi (d0-d4) + GW9662 treatment
SA316885Rosi_GW_2Rosi (d0-d4) + GW9662 treatment
SA316886Rosi_GW_SN4Rosi (d0-d4) + GW9662 treatment supernatant
SA316887Rosi_GW_SN1Rosi (d0-d4) + GW9662 treatment supernatant
SA316888Rosi_GW_SN2Rosi (d0-d4) + GW9662 treatment supernatant
SA316889Rosi_GW_SN3Rosi (d0-d4) + GW9662 treatment supernatant
Showing results 1 to 32 of 32


Collection ID:CO003026
Collection Summary:The SGBS cells were obtained from Prof. Martin Wabitsch laboratory at the University Clinic Ulm. SGBS preadipocytes were differentiated according to the standard protocol described previously (Wabitsch et al., 2001).
Sample Type:Adipose tissue
Storage Conditions:-80℃


Treatment ID:TR003042
Treatment Summary:SGSB preadipocytes were maintained at 37°C and 5% CO2 in 95% humidity. To assess the PPARG-independent effects of DINCH and MINCH on central carbon metabolism, SGBS preadipocytes were exposed to differentiation media containing DINCH (10 µM DINCH+GW) or MINCH (10 µM MINCH+GW) supplemented with the PPARG antagonist GW9662 for 12 days. Irreversible blocking of PPARG prior to treatment was achieved by adding the antagonist 1 hour before the addition of the respective chemical. For comparison, SGBS cells were treated with rosiglitazone (d0-d4) as in the standard protocol but in the presence of GW9662 (Rosi+GW), and SGBS cells were treated with GW9662 only for 12 days (Ctrl+GW). During differentiation, a final concentration of 0.01% (v/v) MeOH and 0.02% (v/v) DMSO was added to all differentiation media. Continuous exposure was mimicked by replacing the cell culture medium every other day. Each treatment was performed in four biological replicates (n=4).

Sample Preparation:

Sampleprep ID:SP003039
Sampleprep Summary:Extraction of intracellular and extracellular metabolites was performed by a 1:1:1 methanol:water:chloroform extraction protocol. For the extraction of intracellular metabolites, the culture medium was removed and the cells were rinsed twice with 1 ml of 0.9% ice-cold NaCl. The rinsing solution was removed, and the metabolism of the cells was stopped by adding 400 µL of MeOH (-20 °C) followed by 400 µL of ice-cold H2O containing 10 µM d6-glutarate. Cells were collected using a cell lifter and 400 µL of chloroform was added. After shaking at 1,400 rpm and 4 °C for 20 min, the extraction mixture was centrifuged at 18,000 g and 4 °C for 5 min. Subsequently, 300 µL volume of the polar upper phase was collected and evaporated to complete dryness. For the extraction of extracellular metabolites, 300 µL of the supernatant was extracted by adding 400 µL MeOH (-20 °C) containing 100 nM MEHP, 100 µL ice-cold H2O containing 40 µM d6-glutarate, and 400 µL chloroform (-20 °C). Subsequent sample preparation was identical to the extraction of intracellular metabolites. Note: After measurement of the samples by LC-MS, the raw AUC values uploaded here were normalized to the internal standard (d6-glutarate, if applicable) and DNA content per well (measured by DAPI fluorescence). After normalization, log2 fold changes were calculated by dividing the normalized peak area from each replicate of each treatment by the normalized peak area from each control. Insulin data were not normalized to DAPI because fold changes were calculated by dividing the intensities of the insulin-stimulated cells by the noninsulin-stimulated cells from each treatment.

Combined analysis:

Analysis ID AN004790
Analysis type MS
Chromatography type Reversed phase
Chromatography system Agilent 1290 Infinity II
Column Waters Xselect XP HSS T3 (150 x 2.1mm, 2.5µm)
MS instrument type QTRAP
MS instrument name ABI Sciex 6500+
Units Peak AUC


Chromatography ID:CH003621
Instrument Name:Agilent 1290 Infinity II
Column Name:Waters Xselect XP HSS T3 (150 x 2.1mm, 2.5µm)
Column Temperature:40
Flow Gradient:0-5 min 0% B, 5-9 min 0%- 2% B, 9-9.5 min 2-6% B, 9.5-11.5 min 6% B, 11.5-12 min 6-11% B, 12-13.5 min 11% B, 13.5-15.5 min 11-28% B, 15.5-16.5 min 28-53% B, 16.5-22.5 53% B, 22.5-23 min 53-0% B, 23-33 min 0% B
Flow Rate:0-15.5 min 0.4 mL/min, 15.5-16.5 min 0.4-0.15 mL/min, 16.5-23 min 0.15 mL/min, 23-27 min 0.15-0.4 mL/min, 27-33 min 0.4 mL/min
Solvent A:10mM TBA, 10mM acetic acid, 5% MeOH, 2% IPA in water
Solvent B:100% IPA
Chromatography Type:Reversed phase


MS ID:MS004536
Analysis ID:AN004790
Instrument Name:ABI Sciex 6500+
Instrument Type:QTRAP
MS Comments:For identification and quantitation, a scheduled MRM method was used, with specific transitions for every metabolite. Data acquisition and peak integration were performed in Analyst® software (Version 1.7.1).