ONTOLOGY SOURCE REFERENCE
Term Source Name	"OBI"	"MTBLS"	"EFO"	"OMIT"	"MS"	"NCIT"	"CHMO"
Term Source File	"http://data.bioontology.org/ontologies/OBI"	"https://www.ebi.ac.uk/metabolights/"	""	""	""	""	""
Term Source Version	"29"	"1"	""	""	""	""	""
Term Source Description	"Ontology for Biomedical Investigations"	"Metabolights Ontology"	""	""	""	""	""
INVESTIGATION
Investigation Identifier	"MTBLS4861"
Investigation Title	"Investigation"
Investigation Description	"Created using the MetaboLights Online Editor (MOE)"
Investigation Submission Date	"2022-05-11"
Investigation Public Release Date	"2023-03-06"
Comment[Created With Configuration]	"MetaboLightsConfig20150707"
Comment[Last Opened With Configuration]	"MetaboLightsConfig20150707"
INVESTIGATION PUBLICATIONS
Investigation PubMed ID	""
Investigation Publication DOI	""
Investigation Publication Author List	""
Investigation Publication Title	""
Investigation Publication Status	""
Investigation Publication Status Term Accession Number	""
Investigation Publication Status Term Source REF	""
INVESTIGATION CONTACTS
Investigation Person Last Name	""
Investigation Person First Name	""
Investigation Person Mid Initials	""
Investigation Person Email	""
Investigation Person Phone	""
Investigation Person Fax	""
Investigation Person Address	""
Investigation Person Affiliation	""
Investigation Person Roles	""
Investigation Person Roles Term Accession Number	""
Investigation Person Roles Term Source REF	""
STUDY
Study Identifier	"MTBLS4861"
Study Title	"Profiling metabolome of uterine fluid for early detection of ovarian cancer"
Study Description	"<p>Ovarian cancer causes high mortality with the poorest prognosis in women among gynecological malignancies. Because of the absence of specific early symptoms, most patients are diagnosed at their late stages. Although a few biomarkers such as cancer antigen 125 and human epididymis protein 4 are used for ovarian cancer diagnosis, their low sensitivity and specificity cannot meet the demand of early detection. Neighboring ovarian surface and fallopian tube, uterine cavity pools secreted fluid enriched with amino acids, lipids, proteins, etc, potentially reflecting the status of adjacent organs with minimal interference from abdominal viscera. This study not only reveals metabolic features in uterine fluid of gynecological patients, but also establishes a newly noninvasive approach for early diagnosis of ovarian cancer.</p>"
Study Submission Date	"2022-05-11"
Study Public Release Date	"2023-03-06"
Study File Name	"s_MTBLS4861.txt"
STUDY DESIGN DESCRIPTORS
Study Design Type	"Biomarker"	"cancer"	"untargeted metabolites"	"ultra-performance liquid chromatography-mass spectrometry"
Study Design Type Term Accession Number	"http://purl.obolibrary.org/obo/NCIT_C16342"	"http://www.ebi.ac.uk/efo/EFO_0000311"	"http://www.ebi.ac.uk/metabolights/ontology/MTBLS_000279"	"http://purl.obolibrary.org/obo/CHMO_0000715"
Study Design Type Term Source REF	"NCIT"	"EFO"	"MTBLS"	"CHMO"
STUDY PUBLICATIONS
Study PubMed ID	""
Study Publication DOI	""
Study Publication Author List	"Pan Wang, Jihong Ma, Wenjing Li, Qilong Wang, Yinan Xiao, Yuening Jiang, Xiaoyang Gu, Yu Wu, Suwei Dong, Hongyan Guo, Mo Li"
Study Publication Title	"Profiling metabolome of uterine fluid for early detection of ovarian cancer"
Study Publication Status	"Submitted"
Study Publication Status Term Accession Number	"http://www.ebi.ac.uk/efo/EFO_0001794"
Study Publication Status Term Source REF	"EFO"
STUDY FACTORS
Study Factor Name	"Disease"	"Batch"
Study Factor Type	"disease"	"block"
Study Factor Type Term Accession Number	"http://www.ebi.ac.uk/efo/EFO_0000408"	"http://www.ebi.ac.uk/efo/EFO_0005067"
Study Factor Type Term Source REF	"EFO"	"EFO"
STUDY ASSAYS
Study Assay File Name	"a_MTBLS4861_LC-MS_negative_reverse-phase_metabolite_profiling.txt"	"a_MTBLS4861_LC-MS_positive_hilic_metabolite_profiling.txt"	"a_MTBLS4861_LC-MS_positive_reverse-phase_metabolite_profiling.txt"
Study Assay Measurement Type	"metabolite profiling"	"metabolite profiling"	"metabolite profiling"
Study Assay Measurement Type Term Accession Number	"http://purl.obolibrary.org/obo/OBI_0000366"	"http://purl.obolibrary.org/obo/OBI_0000366"	"http://purl.obolibrary.org/obo/OBI_0000366"
Study Assay Measurement Type Term Source REF	"OBI"	"OBI"	"OBI"
Study Assay Technology Type	"mass spectrometry"	"mass spectrometry"	"mass spectrometry"
Study Assay Technology Type Term Accession Number	"http://purl.obolibrary.org/obo/OBI_0000470"	"http://purl.obolibrary.org/obo/OBI_0000470"	"http://purl.obolibrary.org/obo/OBI_0000470"
Study Assay Technology Type Term Source REF	"OBI"	"OBI"	"OBI"
Study Assay Technology Platform	"Liquid Chromatography MS - negative - reverse phase"	"Liquid Chromatography MS - positive - hilic"	"Liquid Chromatography MS - positive - reverse phase"
STUDY PROTOCOLS
Study Protocol Name	"Sample collection"	"Extraction"	"Chromatography"	"Mass spectrometry"	"Data transformation"	"Metabolite identification"
Study Protocol Type	"Sample collection"	"Extraction"	"Chromatography"	"Mass spectrometry"	"Data transformation"	"Metabolite identification"
Study Protocol Type Term Accession Number	""	""	""	""	""	""
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Study Protocol Description	"<p>Uterine flushing was performed according to previous protocols with minor modification<strong>[1][2]</strong>. With patients in the lithotomy position, the cervix was exposed by a sterile speculum and cleansed with sterile normal saline solution. A size 10 Foley catheter was placed in uterine cavity through the cervix and secured in place by inflating the balloon with 1 mL normal saline, followed by applying gentle traction to seal off the junction between uterine cavity and the cervical canal. Then 2 mL normal saline was slowly infused into uterine cavity through the catheter, left for 30 s before using gentle suction to harvest the fluid, and this procedure was repeated 5x times to ensure a thorough sampling of uterine cavity. The fluid collected each time was pooled together, subsequently aliquoted into 1.5 mL microcentrifuge tubes and stored at -80 °C. For each patient, multiple aliquots of equal amount (100 mL) of uterine fluid were used for metabolomics experiments and targeted tests.</p><p><br></p><p><strong>Refs:</strong></p><p><strong>[1]</strong> Li TC, Ling E, Dalton C, Bolton AE, Cooke ID. Concentration of endometrial protein PP14 in uterine flushings throughout the menstrual cycle in normal, fertile women. Br J Obstet Gynaecol. 1993 May;100(5):460-4. doi:10.1111/j.1471-0528.1993.tb15272.x. PMID:8518247.</p><p><strong>[2]</strong> Xiao Y, Sun X, Yang X, Zhang J, Xue Q, Cai B, Zhou Y. Leukemia inhibitory factor is dysregulated in the endometrium and uterine flushing fluid of patients with adenomyosis during implantation window. Fertil Steril. 2010 Jun;94(1):85-9. doi:10.1016/j.fertnstert.2009.03.012. Epub 2009 Apr 9. PMID:19361790.</p>"	"<p>After thawing on ice at 4 °C for 30 min, 100 mL of uterine fluid samples was mixed with 100 mL methanol. After vortex for 15 s, the mixture was centrifuged at 12,000 rpm for 10 min at 4 °C for protein precipitation. A total of 80 mL of supernatants was collected before analysis. The aim of aqueous methanol extraction was to remove protein fractions while maximizing small molecule recovery. Samples from each patient were prepared in triplicate using the above method, as the resulting extracts were analyzed using reverse-phase liquid chromatography (RPLC)-mass spectrometry (MS) with electrospray ionization (ESI) positive and ESI negative modes, hydrophilic interaction liquid chromatography (HILIC)-MS with ESI positive mode. Quality control (QC) samples were prepared by mixing equal volumes of each patient’s uterine fluid sample.</p>"	"<p>Uterine fluid samples of patients were subjected to untargeted metabolomics conducted by an Ultimate 3000 Rapid Separation Liquid Chromatography (RSLC) system coupled to a Q Exactive hybrid Quadrupole-Orbitrap mass spectrometer (Thermo Scientific). For RP chromatographic separation method, the extract was gradient eluted from a C18 column (Waters ACQUITY UPLC HSS T3, 2.1 mm x 100 mm, 1.8 µm) using mobile phases including 0.1% formic acid in water (phase A) and 0.1% formic acid in methanol (phase B). The gradient program was as follows: 0­.0-1.0 min, 2% B; 1­­.0-7.0 min, 2-100% B; 7.0-14.5 min, 100% B; 14.5-14.6 min, 100-2% B; 14.6-16.0 min, 2% B. For HILIC separation method, the extract was eluted from a HILIC column (Waters UPLC BEH Amide, 2.1 mm x 100 mm, 1.7 µm) using a gradient consisting of 5 mM ammonium formate and 0.1% formic acid in acetonitrile:water (95:5) (phase A) and 0.1% formic acid in water (phase B). Elution was performed with a flow rate of 0.3 mL/min with a gradient spanning from 5% B to 50% B over 7 min. After a pre-run time of 1 min at 5% B, phase B increase from 5% to 50% within 7 min. Next 50% B was kept constant for 1 min with fast decrease to 5% B within 0.1 min. The column temperature was maintained at 40 °C. The flow rate was 0.3 mL/min and the injection volume was 4 mL. A total of 5 QC samples were continuously injected at the start of the run. Throughout the analytical run, 1x QC sample was injected at a regular interval of 8x samples to monitor the platform variability.</p>"	"<p>The full-scan MS analysis was carried out on a Q Exactive hybrid quadrupole Orbitrap mass spectrometer (Thermo Scientific) equipped with a HESI-II probe, which was operated in positive and negative mode at a spray voltage of 3.7 and 3.5 kV, respectively. Other MS parameters were as follows: resolution of 70,000, auto gain control target under 1E+06, maximum isolation time of 50 ms and scan range of 150-1500 m/z.</p>"	"<p>MS data were acquired using <strong>Xcalibur 2.2 SP1.48</strong> software (Thermo Scientific). MS raw data were processed with <strong>Progenesis QI</strong> (Nonlinear dynamics, version 2.1) for peak alignment, peak picking, deconvolution and eventual feature (m/z-retention time pairs) database searching.</p>"	"<p>Mass spectrometric peaks were preliminarily annotated using the <strong>human metabolome database</strong> (<strong>HMDB; </strong><a href='https://hmdb.ca/' rel='noopener noreferrer' target='_blank'>https://hmdb.ca/</a>)<strong>[1]</strong> with precursor and fragmentation ion accuracy set to 10 ppm. Metabolites were filtered according to the 80% rule<strong>[2]</strong>, i.e., the metabolite was included for further analysis only if it was detected in at least 4/5 samples of one group. The remaining missing values were replaced by 1/5 of the minimal value of the corresponding metabolites. To eliminate the influence of varied metabolite dilution related to the uterine fluid collection volume, the metabolite abundance was normalized by dividing the peak area of each metabolite by the sum of all peak areas of all metabolites<strong>[3]</strong>. Then the data was processed by log10 transformation and Pareto scaling.</p><p><br></p><p><strong>Refs:</strong></p><p><strong>[1]</strong> Wishart DS, Feunang YD, Marcu A, Guo AC, Liang K, Vázquez-Fresno R, Sajed T, Johnson D, Li C, Karu N, Sayeeda Z, Lo E, Assempour N, Berjanskii M, Singhal S, Arndt D, Liang Y, Badran H, Grant J, Serra-Cayuela A, Liu Y, Mandal R, Neveu V, Pon A, Knox C, Wilson M, Manach C, Scalbert A. HMDB 4.0: the human metabolome database for 2018. Nucleic Acids Res. 2018 Jan 4;46(D1):D608-D617. doi:10.1093/nar/gkx1089. PMID:29140435.</p><p><strong>[2]</strong> Bijlsma S, Bobeldijk I, Verheij ER, Ramaker R, Kochhar S, Macdonald IA, van Ommen B, Smilde AK. Large-scale human metabolomics studies: a strategy for data (pre-) processing and validation. Anal Chem. 2006 Jan 15;78(2):567-74. doi:10.1021/ac051495j. PMID:16408941.</p><p><strong>[3]</strong> Chan EC, Pasikanti KK, Nicholson JK. Global urinary metabolic profiling procedures using gas chromatography-mass spectrometry. Nat Protoc. 2011 Sep 8;6(10):1483-99. doi:10.1038/nprot.2011.375. PMID:21959233.</p>"
Study Protocol URI	""	""	""	""	""	""
Study Protocol Version	""	""	""	""	""	""
Study Protocol Parameters Name	""	"Post Extraction;Derivatization"	"Chromatography Instrument;Autosampler model;Column model;Column type;Guard column"	"Scan polarity;Scan m/z range;Instrument;Ion source;Mass analyzer"	""	""
Study Protocol Parameters Name Term Accession Number	""	";"	";;;;"	";;;;"	""	""
Study Protocol Parameters Name Term Source REF	""	";"	";;;;"	";;;;"	""	""
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Study Protocol Components Type Term Accession Number	""	""	""	""	""	""
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STUDY CONTACTS
Study Person Last Name	"Wang"
Study Person First Name	"Pan"
Study Person Mid Initials	""
Study Person Email	"lenord2015@outlook.com"
Study Person Phone	""
Study Person Fax	""
Study Person Address	""
Study Person Affiliation	""
Study Person Roles	"first author"
Study Person Roles Term Accession Number	"http://purl.obolibrary.org/obo/MS_1002034"
Study Person Roles Term Source REF	"MS"
