Summary of Study ST003542

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002178. The data can be accessed directly via it's Project DOI: 10.21228/M83C2C This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

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.

Show all samples  |  Perform analysis on untargeted data  
Download mwTab file (text)   |  Download mwTab file(JSON)   |  Download data files (Contains raw data)
Study IDST003542
Study TitleDysregulated Follicular Fluid Metabolism in Women with Unexplained Infertility
Study TypeHILIC and RPLC based Untargeted LC-MS/MS
Study SummaryResearch Question: Are there specific metabolomic alterations in the Follicular fluid (FF) of women with unexplained infertility (UI)? Study Design: This case-control study included 20 women undergoing in-vitro fertilization (IVF), comparing 10 women diagnosed with UI to 10 control women whose male partners had abnormal semen parameters. FF samples were collected during oocyte retrieval and analysed using hydrophilic interaction liquid chromatography and reversed-phase liquid chromatography, coupled with tandem mass spectrometry (MS/MS) on a Q-TOF mass spectrometer. Metabolites were identified using XCMS Online and MetaboAnalyst, followed by pathway enrichment analysis via the KEGG database. Statistical analyses including OPLS-DA and ROC analysis assessed their diagnostic potential. Metabolite levels were correlated with clinical parameters, including embryo development rates, oocyte maturation, and IVF outcomes. Results: In women with UI, 12 metabolites, including Diacylglycerols, Phosphatidic acids, Vitamin D3 glucosiduronate, 1α-hydroxy-2β-(5-hydroxypentoxy) vitamin D3, Asparginyl-Asparagine, Lithocholic acid, Leu-Pro-Ala-Ser-Phe, Triacylglycerols, Phosphatidyl choline, Phosphatidylethanolamine and Lactosyl ceramide were significantly decreased, while Ile-Lys-Val-Val was significantly increased compared to controls. These metabolites were linked to glycerophospholipid metabolism, glycerolipid metabolism, and steroid synthesis pathways. PLS-DA, OPLS-DA, and ROC analysis indicated high diagnostic performance, with AUC values exceeding 0.8. Additionally, Vitamin D3 glucosiduronate levels negatively correlated with embryo development rates, while Asparginyl-Asparagine levels demonstrated a positive correlation with the MII oocyte rate. Conclusion: This study constitutes the first comprehensive characterization of metabolic dysregulation in the FF of women with UI, offering novel insights into the underlying mechanisms contributing to this condition and advocates for routine assessment of vitamin D3 levels in serum/FF of these women.
Institute
ICMR - National Institute for Research in Reproductive and Child Health
DepartmentGamete Immunobiology
Last NamePanchal
First NameDurva
AddressICMR - NIRRCH, J.M.Street, Parel, Mumbai, India
Emaildurva_gib@nirrh.res.in
Phone9619565514
Submit Date2024-10-25
Num Groups2
Total Subjects20
Num Females20 (10 per group)
Study CommentsThis case-control study included 20 women undergoing in-vitro fertilization (IVF), comprising 10 women diagnosed with unexplained infertility (UI) and 10 control women whose male partners exhibited abnormal semen parameters. The aim was to investigate metabolic differences in the follicular fluid of these groups, contributing to a deeper understanding of factors influencing reproductive outcomes.
PublicationsNone
Raw Data AvailableYes
Raw Data File Type(s)mzML
Analysis Type DetailLC-MS
Release Date2024-11-20
Release Version1
Durva Panchal Durva Panchal
https://dx.doi.org/10.21228/M83C2C
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR002178
Project DOI:doi: 10.21228/M83C2C
Project Title:Dysregulated Follicular Fluid Metabolism in Women with Unexplained Infertility
Project Type:HILIC and RPLC-Based Untargeted LC-MS/MS
Project Summary:Research Question: Are there specific metabolomic alterations in the Follicular fluid (FF) of women with unexplained infertility (UI)? Study Design: This case-control study included 20 women undergoing in-vitro fertilization (IVF), comparing 10 women diagnosed with UI to 10 control women whose male partners had abnormal semen parameters. FF samples were collected during oocyte retrieval and analysed using hydrophilic interaction liquid chromatography and reversed-phase liquid chromatography, coupled with tandem mass spectrometry (MS/MS) on a Q-TOF mass spectrometer. Metabolites were identified using XCMS Online and MetaboAnalyst, followed by pathway enrichment analysis via the KEGG database. Statistical analyses including OPLS-DA and ROC analysis assessed their diagnostic potential. Metabolite levels were correlated with clinical parameters, including embryo development rates, oocyte maturation, and IVF outcomes. Results: In women with UI, 12 metabolites, including Diacylglycerols, Phosphatidic acids, Vitamin D3 glucosiduronate, 1α-hydroxy-2β-(5-hydroxypentoxy) vitamin D3, Asparginyl-Asparagine, Lithocholic acid, Leu-Pro-Ala-Ser-Phe, Triacylglycerols, Phosphatidyl choline, Phosphatidylethanolamine and Lactosyl ceramide were significantly decreased, while Ile-Lys-Val-Val was significantly increased compared to controls. These metabolites were linked to glycerophospholipid metabolism, glycerolipid metabolism, and steroid synthesis pathways. PLS-DA, OPLS-DA, and ROC analysis indicated high diagnostic performance, with AUC values exceeding 0.8. Additionally, Vitamin D3 glucosiduronate levels negatively correlated with embryo development rates, while Asparginyl-Asparagine levels demonstrated a positive correlation with the MII oocyte rate. Conclusion: This study constitutes the first comprehensive characterization of metabolic dysregulation in the FF of women with UI, offering novel insights into the underlying mechanisms contributing to this condition and advocates for routine assessment of vitamin D3 levels in serum/FF of these women.
Institute:ICMR - National Institute for Research in Reproductive and Child Health
Department:Gamete Immunobiology
Last Name:Panchal
First Name:Durva
Address:J.M. Street, Parel, Mumbai, Maharashtra, 400012, India
Email:durva_gib@nirrh.res.in
Phone:022-2419-2005

Subject:

Subject ID:SU003671
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Age Or Age Range:22 - 40 years
Gender:Female
Species Group:Mammals

Factors:

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

mb_sample_id local_sample_id Fertility status
SA387613R_Control-9Control
SA387614H_Control-ve-9Control
SA387615H_Control-ve-10Control
SA387616H_Control-2Control
SA387617R_Control-2Control
SA387618R_Control-3Control
SA387619R_Control-4Control
SA387620R_Control-5Control
SA387621R_Control-6Control
SA387622R_Control-7Control
SA387623R_Control-8Control
SA387624R_Control-10Control
SA387625H_Control-ve-7Control
SA387626R_Control-ve-1Control
SA387627R_Control-ve-2Control
SA387628R_Control-ve-3Control
SA387629R_Control-ve-4Control
SA387630R_Control-ve-5Control
SA387631R_Control-ve-6Control
SA387632R_Control-ve-7Control
SA387633R_Control-ve-8Control
SA387634R_Control-ve-9Control
SA387635R_Control-ve-10Control
SA387636H_Control-ve-8Control
SA387637H_Control-1Control
SA387638H_Control-ve-6Control
SA387639H_Control-ve-2Control
SA387640H_Control-3Control
SA387641H_Control-4Control
SA387642H_Control-5Control
SA387643H_Control-6Control
SA387644H_Control-7Control
SA387645H_Control-8Control
SA387646H_Control-9Control
SA387647H_Control-10Control
SA387648H_Control-ve-5Control
SA387649H_Control-ve-1Control
SA387650R_Control-1Control
SA387651H_Control-ve-4Control
SA387652H_Control-ve-3Control
SA387653H_UI-ve-10UI
SA387654R_UI-ve-1UI
SA387655H_UI-ve-5UI
SA387656H_UI-ve-4UI
SA387657H_UI-ve-3UI
SA387658H_UI-ve-2UI
SA387659H_UI-ve-1UI
SA387660R_UI-ve-2UI
SA387661H_UI-1UI
SA387662R_UI-ve-3UI
SA387663R_UI-ve-4UI
SA387664R_UI-ve-5UI
SA387665R_UI-ve-6UI
SA387666R_UI-ve-7UI
SA387667R_UI-ve-8UI
SA387668R_UI-ve-9UI
SA387669R_UI-ve-10UI
SA387670H_UI-ve-6UI
SA387671H_UI-2UI
SA387672H_UI-ve-9UI
SA387673H_UI-3UI
SA387674H_UI-ve-8UI
SA387675H_UI-10UI
SA387676H_UI-9UI
SA387677H_UI-8UI
SA387678H_UI-7UI
SA387679H_UI-6UI
SA387680H_UI-5UI
SA387681H_UI-4UI
SA387682R_UI-1UI
SA387683R_UI-10UI
SA387684R_UI-2UI
SA387685R_UI-3UI
SA387686H_UI-ve-7UI
SA387687R_UI-5UI
SA387688R_UI-6UI
SA387689R_UI-7UI
SA387690R_UI-8UI
SA387691R_UI-9UI
SA387692R_UI-4UI
Showing results 1 to 80 of 80

Collection:

Collection ID:CO003664
Collection Summary:All participants underwent controlled ovulation stimulation using GnRH antagonist protocols (short-term) and recombinant follicle stimulating hormone (FSH), tailored to their individual requirements. Follicular development and serum sex hormone levels (estradiol, progesterone) were monitored periodically throughout stimulation to assess response and adjust medication dosages as needed. Upon confirmation of at least two follicles exceeding 18 mm in diameter in the bilateral ovaries through ultrasound monitoring, 10,000 IU of human chorionic gonadotropin (HCG) was administered intramuscularly to trigger final oocyte maturation. Oocyte retrieval and FF aspiration were performed transvaginally under ultrasound guidance approximately 34-35 h post HCG injection. During the initial puncture of each follicle exceeding 18mm in diameter, approximately 1 mL of FF was collected in sterile 1.5mL vials (Tarsons, India) under ultrasound guidance and using a sterile aspiration needle. Care was taken to avoid blood contamination of the FF samples and contaminated samples, if any, were excluded. The samples were centrifuged at 12,000 rpm for 20 min at 4°C to eliminate cell debris, aliquoted and cryopreserved at -80°C until analysis.
Collection Protocol Filename:Study_participants_and_Collection_protocol.pdf
Sample Type:Follicular fluid

Treatment:

Treatment ID:TR003680
Treatment Summary:No experimental interventions were administered in this study; rather, we focused on the characterization of endogenous metabolic profiles present in the follicular fluid (FF) of the participants. This approach allowed us to investigate the metabolic landscape and its potential implications on reproductive health

Sample Preparation:

Sampleprep ID:SP003678
Sampleprep Summary:To 40 µg of total protein from each FF sample (UI-and control-group), five volumes of pre-chilled (-20°C) chloroform-methanol solution (2:1 v/v) was added. The mixture was vigorously vortexed for 30 sec to ensure proper mixing and then incubated overnight at 4°C to allow for complete protein precipitation and metabolite partitioning. After overnight incubation, the samples were centrifuged at 12,000 x g for 10 min at 4°C. The upper aqueous fraction, enriched with hydrophilic metabolites, and the bottom chloroform fraction, containing the hydrophobic metabolites, were carefully separated, vacuum dried, and stored at −80°C until use (Nakayasu et al., 2016). Prior to LC-MS/MS analysis, the dried samples were reconstituted in 20μL of a methanol-water solution (1:1 v/v). Each reconstituted sample was then spiked with 2 ppm reserpine (internal standard).
Sampleprep Protocol Filename:Sample_prepartion_for_LCMSMS.pdf

Chromatography:

Chromatography ID:CH004417
Chromatography Summary:Hydrophilic Interaction Liquid Chromatography (HILIC) is a specialized chromatographic technique primarily used for the separation of polar and hydrophilic compounds.
Methods Filename:LC-MSMS_protocol.pdf
Instrument Name:Agilent 6550
Column Name:Thermo Hypersil GOLD (100 x 2.1mm, 3um)
Column Temperature:35
Flow Gradient:Gradient for B: time 0; 1%, 2min; 1%, 8min; 55%, 9min; 99%, 9.1min; 99%, 11min; 99%, 11.1min; 1%, 19min; 1%, 19.1min; 1%, 23min; 1%
Flow Rate:0.3 μL/min
Solvent A:95% acetonitrile / 5% water; 10mM ammonium ammonium formate
Solvent B:30% acetonitrile / 70% water; 10mM ammonium ammonium formate
Chromatography Type:HILIC
  
Chromatography ID:CH004418
Chromatography Summary:Reversed-Phase Liquid Chromatography (RPLC) is a widely used chromatographic technique for the separation of nonpolar and moderately polar compounds based on their hydrophobicity.
Methods Filename:LC-MSMS_protocol.pdf
Instrument Name:Agilent 6550
Column Name:Thermo Syncronis C-18 (150 x 4mm; 5um)
Column Temperature:35
Flow Gradient:Gradient for B:Gradient for B: time 0; 1%, 2min; 1%, 8min; 55%, 9min; 99%, 9.1min; 99%, 11min; 99%, 11.1min; 1%, 19min; 1%, 19.1min; 1%, 23min; 1%
Flow Rate:0.3 μL/min
Solvent A:100% water, 0.1% formic acid
Solvent B:100% methanol; 0.1% formic acid
Chromatography Type:Reversed phase

Analysis:

Analysis ID:AN005814
Analysis Type:MS
Analysis Protocol File:LC-MSMS_Data_analysis.pdf
Chromatography ID:CH004417
Has Mz:1
Has Rt:1
Rt Units:Minutes
Results File:ST003542_AN005814_Results.txt
Units:Intensities
  
Analysis ID:AN005815
Analysis Type:MS
Analysis Protocol File:LC-MSMS_Data_analysis.pdf
Chromatography ID:CH004417
Has Mz:1
Has Rt:1
Rt Units:Minutes
Results File:ST003542_AN005815_Results.txt
Units:Intensities
  
Analysis ID:AN005816
Analysis Type:MS
Analysis Protocol File:LC-MSMS_Data_analysis.pdf
Chromatography ID:CH004418
Has Mz:1
Has Rt:1
Rt Units:Minutes
Results File:ST003542_AN005816_Results.txt
Units:Intensities
  
Analysis ID:AN005817
Analysis Type:MS
Analysis Protocol File:LC-MSMS_Data_analysis.pdf
Chromatography ID:CH004418
Has Mz:1
Has Rt:1
Rt Units:Minutes
Results File:ST003542_AN005817_Results.txt
Units:Intensities
  logo