#METABOLOMICS WORKBENCH Durva_Panchal_20241025_024317 DATATRACK_ID:5315 STUDY_ID:ST003542 ANALYSIS_ID:AN005816 PROJECT_ID:PR002178
VERSION             	1
CREATED_ON             	October 29, 2024, 9:43 am
#PROJECT
PR:PROJECT_TITLE                 	Dysregulated Follicular Fluid Metabolism in Women with Unexplained Infertility
PR:PROJECT_TYPE                  	HILIC and RPLC-Based Untargeted LC-MS/MS
PR:PROJECT_SUMMARY               	Research Question: Are there specific metabolomic alterations in the Follicular
PR:PROJECT_SUMMARY               	fluid (FF) of women with unexplained infertility (UI)? Study Design: This
PR:PROJECT_SUMMARY               	case-control study included 20 women undergoing in-vitro fertilization (IVF),
PR:PROJECT_SUMMARY               	comparing 10 women diagnosed with UI to 10 control women whose male partners had
PR:PROJECT_SUMMARY               	abnormal semen parameters. FF samples were collected during oocyte retrieval and
PR:PROJECT_SUMMARY               	analysed using hydrophilic interaction liquid chromatography and reversed-phase
PR:PROJECT_SUMMARY               	liquid chromatography, coupled with tandem mass spectrometry (MS/MS) on a Q-TOF
PR:PROJECT_SUMMARY               	mass spectrometer. Metabolites were identified using XCMS Online and
PR:PROJECT_SUMMARY               	MetaboAnalyst, followed by pathway enrichment analysis via the KEGG database.
PR:PROJECT_SUMMARY               	Statistical analyses including OPLS-DA and ROC analysis assessed their
PR:PROJECT_SUMMARY               	diagnostic potential. Metabolite levels were correlated with clinical
PR:PROJECT_SUMMARY               	parameters, including embryo development rates, oocyte maturation, and IVF
PR:PROJECT_SUMMARY               	outcomes. Results: In women with UI, 12 metabolites, including Diacylglycerols,
PR:PROJECT_SUMMARY               	Phosphatidic acids, Vitamin D3 glucosiduronate,
PR:PROJECT_SUMMARY               	1α-hydroxy-2β-(5-hydroxypentoxy) vitamin D3, Asparginyl-Asparagine,
PR:PROJECT_SUMMARY               	Lithocholic acid, Leu-Pro-Ala-Ser-Phe, Triacylglycerols, Phosphatidyl choline,
PR:PROJECT_SUMMARY               	Phosphatidylethanolamine and Lactosyl ceramide were significantly decreased,
PR:PROJECT_SUMMARY               	while Ile-Lys-Val-Val was significantly increased compared to controls. These
PR:PROJECT_SUMMARY               	metabolites were linked to glycerophospholipid metabolism, glycerolipid
PR:PROJECT_SUMMARY               	metabolism, and steroid synthesis pathways. PLS-DA, OPLS-DA, and ROC analysis
PR:PROJECT_SUMMARY               	indicated high diagnostic performance, with AUC values exceeding 0.8.
PR:PROJECT_SUMMARY               	Additionally, Vitamin D3 glucosiduronate levels negatively correlated with
PR:PROJECT_SUMMARY               	embryo development rates, while Asparginyl-Asparagine levels demonstrated a
PR:PROJECT_SUMMARY               	positive correlation with the MII oocyte rate. Conclusion: This study
PR:PROJECT_SUMMARY               	constitutes the first comprehensive characterization of metabolic dysregulation
PR:PROJECT_SUMMARY               	in the FF of women with UI, offering novel insights into the underlying
PR:PROJECT_SUMMARY               	mechanisms contributing to this condition and advocates for routine assessment
PR:PROJECT_SUMMARY               	of vitamin D3 levels in serum/FF of these women.
PR:INSTITUTE                     	ICMR - National Institute for Research in Reproductive and Child Health
PR:DEPARTMENT                    	Gamete Immunobiology
PR:LAST_NAME                     	Panchal
PR:FIRST_NAME                    	Durva
PR:ADDRESS                       	J.M. Street, Parel, Mumbai, Maharashtra, 400012, India
PR:EMAIL                         	durva_gib@nirrh.res.in
PR:PHONE                         	022-2419-2005
#STUDY
ST:STUDY_TITLE                   	Dysregulated Follicular Fluid Metabolism in Women with Unexplained Infertility
ST:STUDY_TYPE                    	HILIC and RPLC based Untargeted LC-MS/MS
ST:STUDY_SUMMARY                 	Research Question: Are there specific metabolomic alterations in the Follicular
ST:STUDY_SUMMARY                 	fluid (FF) of women with unexplained infertility (UI)? Study Design: This
ST:STUDY_SUMMARY                 	case-control study included 20 women undergoing in-vitro fertilization (IVF),
ST:STUDY_SUMMARY                 	comparing 10 women diagnosed with UI to 10 control women whose male partners had
ST:STUDY_SUMMARY                 	abnormal semen parameters. FF samples were collected during oocyte retrieval and
ST:STUDY_SUMMARY                 	analysed using hydrophilic interaction liquid chromatography and reversed-phase
ST:STUDY_SUMMARY                 	liquid chromatography, coupled with tandem mass spectrometry (MS/MS) on a Q-TOF
ST:STUDY_SUMMARY                 	mass spectrometer. Metabolites were identified using XCMS Online and
ST:STUDY_SUMMARY                 	MetaboAnalyst, followed by pathway enrichment analysis via the KEGG database.
ST:STUDY_SUMMARY                 	Statistical analyses including OPLS-DA and ROC analysis assessed their
ST:STUDY_SUMMARY                 	diagnostic potential. Metabolite levels were correlated with clinical
ST:STUDY_SUMMARY                 	parameters, including embryo development rates, oocyte maturation, and IVF
ST:STUDY_SUMMARY                 	outcomes. Results: In women with UI, 12 metabolites, including Diacylglycerols,
ST:STUDY_SUMMARY                 	Phosphatidic acids, Vitamin D3 glucosiduronate,
ST:STUDY_SUMMARY                 	1α-hydroxy-2β-(5-hydroxypentoxy) vitamin D3, Asparginyl-Asparagine,
ST:STUDY_SUMMARY                 	Lithocholic acid, Leu-Pro-Ala-Ser-Phe, Triacylglycerols, Phosphatidyl choline,
ST:STUDY_SUMMARY                 	Phosphatidylethanolamine and Lactosyl ceramide were significantly decreased,
ST:STUDY_SUMMARY                 	while Ile-Lys-Val-Val was significantly increased compared to controls. These
ST:STUDY_SUMMARY                 	metabolites were linked to glycerophospholipid metabolism, glycerolipid
ST:STUDY_SUMMARY                 	metabolism, and steroid synthesis pathways. PLS-DA, OPLS-DA, and ROC analysis
ST:STUDY_SUMMARY                 	indicated high diagnostic performance, with AUC values exceeding 0.8.
ST:STUDY_SUMMARY                 	Additionally, Vitamin D3 glucosiduronate levels negatively correlated with
ST:STUDY_SUMMARY                 	embryo development rates, while Asparginyl-Asparagine levels demonstrated a
ST:STUDY_SUMMARY                 	positive correlation with the MII oocyte rate. Conclusion: This study
ST:STUDY_SUMMARY                 	constitutes the first comprehensive characterization of metabolic dysregulation
ST:STUDY_SUMMARY                 	in the FF of women with UI, offering novel insights into the underlying
ST:STUDY_SUMMARY                 	mechanisms contributing to this condition and advocates for routine assessment
ST:STUDY_SUMMARY                 	of vitamin D3 levels in serum/FF of these women.
ST:INSTITUTE                     	ICMR - National Institute for Research in Reproductive and Child Health
ST:DEPARTMENT                    	Gamete Immunobiology
ST:LAST_NAME                     	Panchal
ST:FIRST_NAME                    	Durva
ST:ADDRESS                       	ICMR - NIRRCH, J.M.Street, Parel, Mumbai, India
ST:EMAIL                         	durva_gib@nirrh.res.in
ST:PHONE                         	9619565514
ST:NUM_GROUPS                    	2
ST:TOTAL_SUBJECTS                	20
ST:NUM_FEMALES                   	20 (10 per group)
ST:STUDY_COMMENTS                	This case-control study included 20 women undergoing in-vitro fertilization
ST:STUDY_COMMENTS                	(IVF), comprising 10 women diagnosed with unexplained infertility (UI) and 10
ST:STUDY_COMMENTS                	control women whose male partners exhibited abnormal semen parameters. The aim
ST:STUDY_COMMENTS                	was to investigate metabolic differences in the follicular fluid of these
ST:STUDY_COMMENTS                	groups, contributing to a deeper understanding of factors influencing
ST:STUDY_COMMENTS                	reproductive outcomes.
ST:PUBLICATIONS                  	None
#SUBJECT
SU:SUBJECT_TYPE                  	Human
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
SU:AGE_OR_AGE_RANGE              	22 - 40 years
SU:GENDER                        	Female
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	S1	H_Control-1	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Positive_Sample1.mzML
SUBJECT_SAMPLE_FACTORS           	S2	H_Control-2	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Positive_Sample2.mzML
SUBJECT_SAMPLE_FACTORS           	S3	H_Control-3	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Positive_Sample3.mzML
SUBJECT_SAMPLE_FACTORS           	S4	H_Control-4	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Positive_Sample4.mzML
SUBJECT_SAMPLE_FACTORS           	S5	H_Control-5	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Positive_Sample5.mzML
SUBJECT_SAMPLE_FACTORS           	S6	H_Control-6	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Positive_Sample6.mzML
SUBJECT_SAMPLE_FACTORS           	S7	H_Control-7	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Positive_Sample7.mzML
SUBJECT_SAMPLE_FACTORS           	S8	H_Control-8	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Positive_Sample8.mzML
SUBJECT_SAMPLE_FACTORS           	S9	H_Control-9	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Positive_Sample9.mzML
SUBJECT_SAMPLE_FACTORS           	S10	H_Control-10	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Positive_Sample10.mzML
SUBJECT_SAMPLE_FACTORS           	S11	H_UI-1	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Positive_Sample1.mzML
SUBJECT_SAMPLE_FACTORS           	S12	H_UI-2	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Positive_Sample2.mzML
SUBJECT_SAMPLE_FACTORS           	S13	H_UI-3	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Positive_Sample3.mzML
SUBJECT_SAMPLE_FACTORS           	S14	H_UI-4	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Positive_Sample4.mzML
SUBJECT_SAMPLE_FACTORS           	S15	H_UI-5	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Positive_Sample5.mzML
SUBJECT_SAMPLE_FACTORS           	S16	H_UI-6	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Positive_Sample6.mzML
SUBJECT_SAMPLE_FACTORS           	S17	H_UI-7	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Positive_Sample7.mzML
SUBJECT_SAMPLE_FACTORS           	S18	H_UI-8	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Positive_Sample8.mzML
SUBJECT_SAMPLE_FACTORS           	S19	H_UI-9	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Positive_Sample9.mzML
SUBJECT_SAMPLE_FACTORS           	S20	H_UI-10	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Positive_Sample10.mzML
SUBJECT_SAMPLE_FACTORS           	S21	H_Control-ve-1	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Negative_Sample1.mzML
SUBJECT_SAMPLE_FACTORS           	S22	H_Control-ve-2	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Negative_Sample2.mzML
SUBJECT_SAMPLE_FACTORS           	S23	H_Control-ve-3	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Negative_Sample3.mzML
SUBJECT_SAMPLE_FACTORS           	S24	H_Control-ve-4	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Negative_Sample4.mzML
SUBJECT_SAMPLE_FACTORS           	S25	H_Control-ve-5	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Negative_Sample5.mzML
SUBJECT_SAMPLE_FACTORS           	S26	H_Control-ve-6	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Negative_Sample6.mzML
SUBJECT_SAMPLE_FACTORS           	S27	H_Control-ve-7	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Negative_Sample7.mzML
SUBJECT_SAMPLE_FACTORS           	S28	H_Control-ve-8	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Negative_Sample8.mzML
SUBJECT_SAMPLE_FACTORS           	S29	H_Control-ve-9	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Negative_Sample9.mzML
SUBJECT_SAMPLE_FACTORS           	S30	H_Control-ve-10	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_HILIC_Negative_Sample10.mzML
SUBJECT_SAMPLE_FACTORS           	S31	H_UI-ve-1	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Negative_Sample1.mzML
SUBJECT_SAMPLE_FACTORS           	S32	H_UI-ve-2	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Negative_Sample2.mzML
SUBJECT_SAMPLE_FACTORS           	S33	H_UI-ve-3	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Negative_Sample3.mzML
SUBJECT_SAMPLE_FACTORS           	S34	H_UI-ve-4	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Negative_Sample4.mzML
SUBJECT_SAMPLE_FACTORS           	S35	H_UI-ve-5	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Negative_Sample5.mzML
SUBJECT_SAMPLE_FACTORS           	S36	H_UI-ve-6	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Negative_Sample6.mzML
SUBJECT_SAMPLE_FACTORS           	S37	H_UI-ve-7	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Negative_Sample7.mzML
SUBJECT_SAMPLE_FACTORS           	S38	H_UI-ve-8	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Negative_Sample8.mzML
SUBJECT_SAMPLE_FACTORS           	S39	H_UI-ve-9	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Negative_Sample9.mzML
SUBJECT_SAMPLE_FACTORS           	S40	H_UI-ve-10	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_HILIC_Negative_Sample10.mzML
SUBJECT_SAMPLE_FACTORS           	S41	R_Control-1	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Positive_Sample1.mzML
SUBJECT_SAMPLE_FACTORS           	S42	R_Control-2	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Positive_Sample2.mzML
SUBJECT_SAMPLE_FACTORS           	S43	R_Control-3	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Positive_Sample3.mzML
SUBJECT_SAMPLE_FACTORS           	S44	R_Control-4	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Positive_Sample4.mzML
SUBJECT_SAMPLE_FACTORS           	S45	R_Control-5	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Positive_Sample5.mzML
SUBJECT_SAMPLE_FACTORS           	S46	R_Control-6	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Positive_Sample6.mzML
SUBJECT_SAMPLE_FACTORS           	S47	R_Control-7	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Positive_Sample7.mzML
SUBJECT_SAMPLE_FACTORS           	S48	R_Control-8	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Positive_Sample8.mzML
SUBJECT_SAMPLE_FACTORS           	S49	R_Control-9	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Positive_Sample9.mzML
SUBJECT_SAMPLE_FACTORS           	S50	R_Control-10	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Positive_Sample10.mzML
SUBJECT_SAMPLE_FACTORS           	S51	R_UI-1	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Positive_Sample1.mzML
SUBJECT_SAMPLE_FACTORS           	S52	R_UI-2	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Positive_Sample2.mzML
SUBJECT_SAMPLE_FACTORS           	S53	R_UI-3	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Positive_Sample3.mzML
SUBJECT_SAMPLE_FACTORS           	S54	R_UI-4	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Positive_Sample4.mzML
SUBJECT_SAMPLE_FACTORS           	S55	R_UI-5	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Positive_Sample5.mzML
SUBJECT_SAMPLE_FACTORS           	S56	R_UI-6	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Positive_Sample6.mzML
SUBJECT_SAMPLE_FACTORS           	S57	R_UI-7	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Positive_Sample7.mzML
SUBJECT_SAMPLE_FACTORS           	S58	R_UI-8	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Positive_Sample8.mzML
SUBJECT_SAMPLE_FACTORS           	S59	R_UI-9	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Positive_Sample9.mzML
SUBJECT_SAMPLE_FACTORS           	S60	R_UI-10	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Positive_Sample10.mzML
SUBJECT_SAMPLE_FACTORS           	S61	R_Control-ve-1	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Negative_Sample1.mzML
SUBJECT_SAMPLE_FACTORS           	S62	R_Control-ve-2	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Negative_Sample2.mzML
SUBJECT_SAMPLE_FACTORS           	S63	R_Control-ve-3	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Negative_Sample3.mzML
SUBJECT_SAMPLE_FACTORS           	S64	R_Control-ve-4	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Negative_Sample4.mzML
SUBJECT_SAMPLE_FACTORS           	S65	R_Control-ve-5	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Negative_Sample5.mzML
SUBJECT_SAMPLE_FACTORS           	S66	R_Control-ve-6	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Negative_Sample6.mzML
SUBJECT_SAMPLE_FACTORS           	S67	R_Control-ve-7	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Negative_Sample7.mzML
SUBJECT_SAMPLE_FACTORS           	S68	R_Control-ve-8	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Negative_Sample8.mzML
SUBJECT_SAMPLE_FACTORS           	S69	R_Control-ve-9	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Negative_Sample9.mzML
SUBJECT_SAMPLE_FACTORS           	S70	R_Control-ve-10	Sample source:Follicular fluid | Fertility status:Control	RAW_FILE_NAME(Raw file name)=Control_RPLC_Negative_Sample10.mzML
SUBJECT_SAMPLE_FACTORS           	S71	R_UI-ve-1	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Negative_Sample1.mzML
SUBJECT_SAMPLE_FACTORS           	S72	R_UI-ve-2	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Negative_Sample2.mzML
SUBJECT_SAMPLE_FACTORS           	S73	R_UI-ve-3	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Negative_Sample3.mzML
SUBJECT_SAMPLE_FACTORS           	S74	R_UI-ve-4	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Negative_Sample4.mzML
SUBJECT_SAMPLE_FACTORS           	S75	R_UI-ve-5	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Negative_Sample5.mzML
SUBJECT_SAMPLE_FACTORS           	S76	R_UI-ve-6	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Negative_Sample6.mzML
SUBJECT_SAMPLE_FACTORS           	S77	R_UI-ve-7	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Negative_Sample7.mzML
SUBJECT_SAMPLE_FACTORS           	S78	R_UI-ve-8	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Negative_Sample8.mzML
SUBJECT_SAMPLE_FACTORS           	S79	R_UI-ve-9	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Negative_Sample9.mzML
SUBJECT_SAMPLE_FACTORS           	S80	R_UI-ve-10	Sample source:Follicular fluid | Fertility status:UI	RAW_FILE_NAME(Raw file name)=Unexplained Infertility_RPLC_Negative_Sample10.mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	All participants underwent controlled ovulation stimulation using GnRH
CO:COLLECTION_SUMMARY            	antagonist protocols (short-term) and recombinant follicle stimulating hormone
CO:COLLECTION_SUMMARY            	(FSH), tailored to their individual requirements. Follicular development and
CO:COLLECTION_SUMMARY            	serum sex hormone levels (estradiol, progesterone) were monitored periodically
CO:COLLECTION_SUMMARY            	throughout stimulation to assess response and adjust medication dosages as
CO:COLLECTION_SUMMARY            	needed. Upon confirmation of at least two follicles exceeding 18 mm in diameter
CO:COLLECTION_SUMMARY            	in the bilateral ovaries through ultrasound monitoring, 10,000 IU of human
CO:COLLECTION_SUMMARY            	chorionic gonadotropin (HCG) was administered intramuscularly to trigger final
CO:COLLECTION_SUMMARY            	oocyte maturation. Oocyte retrieval and FF aspiration were performed
CO:COLLECTION_SUMMARY            	transvaginally under ultrasound guidance approximately 34-35 h post HCG
CO:COLLECTION_SUMMARY            	injection. During the initial puncture of each follicle exceeding 18mm in
CO:COLLECTION_SUMMARY            	diameter, approximately 1 mL of FF was collected in sterile 1.5mL vials
CO:COLLECTION_SUMMARY            	(Tarsons, India) under ultrasound guidance and using a sterile aspiration
CO:COLLECTION_SUMMARY            	needle. Care was taken to avoid blood contamination of the FF samples and
CO:COLLECTION_SUMMARY            	contaminated samples, if any, were excluded. The samples were centrifuged at
CO:COLLECTION_SUMMARY            	12,000 rpm for 20 min at 4°C to eliminate cell debris, aliquoted and
CO:COLLECTION_SUMMARY            	cryopreserved at -80°C until analysis.
CO:COLLECTION_PROTOCOL_FILENAME  	Study_participants_and_Collection_protocol.pdf
CO:SAMPLE_TYPE                   	Follicular fluid
#TREATMENT
TR:TREATMENT_SUMMARY             	No experimental interventions were administered in this study; rather, we
TR:TREATMENT_SUMMARY             	focused on the characterization of endogenous metabolic profiles present in the
TR:TREATMENT_SUMMARY             	follicular fluid (FF) of the participants. This approach allowed us to
TR:TREATMENT_SUMMARY             	investigate the metabolic landscape and its potential implications on
TR:TREATMENT_SUMMARY             	reproductive health
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	To 40 µg of total protein from each FF sample (UI-and control-group), five
SP:SAMPLEPREP_SUMMARY            	volumes of pre-chilled (-20°C) chloroform-methanol solution (2:1 v/v) was
SP:SAMPLEPREP_SUMMARY            	added. The mixture was vigorously vortexed for 30 sec to ensure proper mixing
SP:SAMPLEPREP_SUMMARY            	and then incubated overnight at 4°C to allow for complete protein precipitation
SP:SAMPLEPREP_SUMMARY            	and metabolite partitioning. After overnight incubation, the samples were
SP:SAMPLEPREP_SUMMARY            	centrifuged at 12,000 x g for 10 min at 4°C. The upper aqueous fraction,
SP:SAMPLEPREP_SUMMARY            	enriched with hydrophilic metabolites, and the bottom chloroform fraction,
SP:SAMPLEPREP_SUMMARY            	containing the hydrophobic metabolites, were carefully separated, vacuum dried,
SP:SAMPLEPREP_SUMMARY            	and stored at −80°C until use (Nakayasu et al., 2016). Prior to LC-MS/MS
SP:SAMPLEPREP_SUMMARY            	analysis, the dried samples were reconstituted in 20μL of a methanol-water
SP:SAMPLEPREP_SUMMARY            	solution (1:1 v/v). Each reconstituted sample was then spiked with 2 ppm
SP:SAMPLEPREP_SUMMARY            	reserpine (internal standard).
SP:SAMPLEPREP_PROTOCOL_FILENAME  	Sample_prepartion_for_LCMSMS.pdf
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Reversed-Phase Liquid Chromatography (RPLC) is a widely used chromatographic
CH:CHROMATOGRAPHY_SUMMARY        	technique for the separation of nonpolar and moderately polar compounds based on
CH:CHROMATOGRAPHY_SUMMARY        	their hydrophobicity.
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent 6550
CH:COLUMN_NAME                   	Thermo Syncronis C-18 (150 x 4mm; 5um)
CH:SOLVENT_A                     	100% water, 0.1% formic acid
CH:SOLVENT_B                     	100% methanol; 0.1% formic acid
CH:FLOW_GRADIENT                 	Gradient for B:Gradient for B: time 0; 1%, 2min; 1%, 8min; 55%, 9min; 99%,
CH:FLOW_GRADIENT                 	9.1min; 99%, 11min; 99%, 11.1min; 1%, 19min; 1%, 19.1min; 1%, 23min; 1%
CH:FLOW_RATE                     	0.3 μL/min
CH:COLUMN_TEMPERATURE            	35
CH:METHODS_FILENAME              	LC-MSMS_protocol.pdf
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:ANALYSIS_PROTOCOL_FILE        	LC-MSMS_Data_analysis.pdf
#MS
MS:INSTRUMENT_NAME               	Agilent 6550 QTOF
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	Mass spectrometry acquisition was performed using an Agilent 6550 iFunnel Q-TOF
MS:MS_COMMENTS                   	LC/MS system in both positive and negative ionization modes, covering m/z 60 to
MS:MS_COMMENTS                   	1000 at a scan rate of 1 spectrum per second. Data processing utilized XCMS
MS:MS_COMMENTS                   	Online (version 3.7.1) for feature identification and normalization, with raw
MS:MS_COMMENTS                   	files converted to .mzML format via ProteoWizard. Control and unexplained
MS:MS_COMMENTS                   	infertility (UI) group data in .mzML format were subsequently uploaded as
MS:MS_COMMENTS                   	separate datasets to XCMS Online for pairwise comparison. Differentially
MS:MS_COMMENTS                   	expressed metabolites in the follicular fluid (FF) were identified and forwaded
MS:MS_COMMENTS                   	through MetaboAnalyst's "MS Peaks to Pathways" function. The Mummichog algorithm
MS:MS_COMMENTS                   	was employed to identify enriched metabolic pathways using a p-value threshold
MS:MS_COMMENTS                   	of 0.001, with the KEGG database for Homo sapiens serving as the reference for
MS:MS_COMMENTS                   	metabolite annotation and pathway assignment.
MS:MS_RESULTS_FILE               	ST003542_AN005816_Results.txt	UNITS:Intensities	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END