#METABOLOMICS WORKBENCH lleonca_20250522_123227 DATATRACK_ID:5939 STUDY_ID:ST003936 ANALYSIS_ID:AN006478 PROJECT_ID:PR002466
VERSION             	1
CREATED_ON             	June 3, 2025, 9:08 pm
#PROJECT
PR:PROJECT_TITLE                 	Metabolomic analysis of breast cancer in Colombian patients: Exploring molecular
PR:PROJECT_TITLE                 	signatures in different subtypes and stages
PR:PROJECT_SUMMARY               	Breast cancer is a neoplasm characterized by high heterogeneity, influenced by
PR:PROJECT_SUMMARY               	intrinsic molecular subtypes and clinical stage, aspects that remain
PR:PROJECT_SUMMARY               	underexplored in the Colombian population. This study aimed to characterize the
PR:PROJECT_SUMMARY               	metabolic alterations associated with subtypes and disease pro-gression using an
PR:PROJECT_SUMMARY               	untargeted metabolomics approach in a cohort of newly diagnosed,
PR:PROJECT_SUMMARY               	treat-ment-naive Colombian women. Samples were analyzed using LC-QTOF-MS and
PR:PROJECT_SUMMARY               	GC-QTOF-MS, along with amino acid profiling to improve metabolite coverage.
PR:PROJECT_SUMMARY               	Alterations consistent with previous studies were identified; however, the
PR:PROJECT_SUMMARY               	Luminal B subtype showed elevated levels of long-chain acylcarnitines, along
PR:PROJECT_SUMMARY               	with higher concentrations of free fatty acids compared to other subtypes. It
PR:PROJECT_SUMMARY               	also presented elevated levels of carbohydrates and essential glycolytic
PR:PROJECT_SUMMARY               	intermediates, suggesting that this subtype may adopt a hybrid metabolic
PR:PROJECT_SUMMARY               	phenotype characterized by increased glycolytic flux as well as enhanced fatty
PR:PROJECT_SUMMARY               	acid catabolism. TNM (Tumor, Node, Metastasis) staging analysis showed
PR:PROJECT_SUMMARY               	progres-sive metabolic reprogramming of breast cancer. In advanced stages, a
PR:PROJECT_SUMMARY               	sustained increase in phosphatidylcholines and a decrease in
PR:PROJECT_SUMMARY               	lysophosphatidylcholines were observed, reflecting lipid alterations associated
PR:PROJECT_SUMMARY               	with key roles in tumor progression. In early stages (I-II), plasma metabo-lites
PR:PROJECT_SUMMARY               	with high discriminatory power were identified, such as pyrrolidone glutamic
PR:PROJECT_SUMMARY               	acid, ribose, and glycerol, which are associated with dysfunctions in energy and
PR:PROJECT_SUMMARY               	carbohydrate metabolism. These results highlight metabolomics as a promising
PR:PROJECT_SUMMARY               	tool for early diagnosis, clinical follow-up, and molecular characterization in
PR:PROJECT_SUMMARY               	breast cancer.
PR:INSTITUTE                     	Universidad de los Andes, Colombia
PR:LAST_NAME                     	León Carreño, Bogota
PR:FIRST_NAME                    	Lizeth
PR:ADDRESS                       	Carrera 3#21-46
PR:EMAIL                         	l.leonc@uniandes.edu.co
PR:PHONE                         	3138218641
#STUDY
ST:STUDY_TITLE                   	Metabolomic analysis of breast cancer in Colombian patients: Exploring molecular
ST:STUDY_TITLE                   	signatures in different subtypes and stages
ST:STUDY_SUMMARY                 	Breast cancer is a neoplasm characterized by high heterogeneity, influenced by
ST:STUDY_SUMMARY                 	intrinsic molecular subtypes and clinical stage, aspects that remain
ST:STUDY_SUMMARY                 	underexplored in the Colombian population. This study aimed to characterize the
ST:STUDY_SUMMARY                 	metabolic alterations associated with subtypes and disease progression using an
ST:STUDY_SUMMARY                 	untargeted metabolomics approach in a cohort of newly diagnosed, treatment-naive
ST:STUDY_SUMMARY                 	Colombian women. Samples were analyzed using LC-QTOF-MS and GC-QTOF-MS, along
ST:STUDY_SUMMARY                 	with amino acid profiling to improve metabolite coverage. Alterations consistent
ST:STUDY_SUMMARY                 	with previous studies were identified; however, the Luminal B subtype showed
ST:STUDY_SUMMARY                 	elevated levels of long-chain acylcarnitines, along with higher concentrations
ST:STUDY_SUMMARY                 	of free fatty acids compared to other subtypes. It also presented elevated
ST:STUDY_SUMMARY                 	levels of carbohydrates and essential glycolytic intermediates, suggesting that
ST:STUDY_SUMMARY                 	this subtype may adopt a hybrid metabolic phenotype characterized by increased
ST:STUDY_SUMMARY                 	glycolytic flux as well as enhanced fatty acid catabolism. TNM (Tumor, Node,
ST:STUDY_SUMMARY                 	Metastasis) staging analysis showed progres-sive metabolic reprogramming of
ST:STUDY_SUMMARY                 	breast cancer. In advanced stages, a sustained increase in phosphatidylcholines
ST:STUDY_SUMMARY                 	and a decrease in lysophosphatidylcholines were observed, reflecting lipid
ST:STUDY_SUMMARY                 	alterations associated with key roles in tumor progression. In early stages
ST:STUDY_SUMMARY                 	(I-II), plasma metabolites with high discriminatory power were identified, such
ST:STUDY_SUMMARY                 	as pyrrolidone glutamic acid, ribose, and glycerol, which are associated with
ST:STUDY_SUMMARY                 	dysfunctions in energy and carbohydrate metabolism. These results highlight
ST:STUDY_SUMMARY                 	metabolomics as a promising tool for early diagnosis, clinical follow-up, and
ST:STUDY_SUMMARY                 	molecular characterization in breast cancer.
ST:INSTITUTE                     	Universidad de los Andes
ST:LAST_NAME                     	Leon Carreño
ST:FIRST_NAME                    	Lizeth
ST:ADDRESS                       	Carrera 3#21-46, Bogota
ST:EMAIL                         	l.leonc@uniandes.edu.co
ST:PHONE                         	3138218641
#SUBJECT
SU:SUBJECT_TYPE                  	Human
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
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           	-	M002	Subtype:Luminal B | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M002.mzML
SUBJECT_SAMPLE_FACTORS           	-	M005	Subtype:Luminal B | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M005.mzML
SUBJECT_SAMPLE_FACTORS           	-	M009	Subtype:HER2 | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M009.mzML
SUBJECT_SAMPLE_FACTORS           	-	M012	Subtype:Luminal A | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M012.mzML
SUBJECT_SAMPLE_FACTORS           	-	M014	Subtype:Basal like | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M014.mzML
SUBJECT_SAMPLE_FACTORS           	-	M015	Subtype:Luminal A | Stage:NA | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M015.mzML
SUBJECT_SAMPLE_FACTORS           	-	M017	Subtype:Luminal A | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M017.mzML
SUBJECT_SAMPLE_FACTORS           	-	M020	Subtype:Basal like | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M020.mzML
SUBJECT_SAMPLE_FACTORS           	-	M023	Subtype:Luminal A | Stage:I | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M023.mzML
SUBJECT_SAMPLE_FACTORS           	-	M027	Subtype:Luminal B | Stage:NA | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M027.mzML
SUBJECT_SAMPLE_FACTORS           	-	M028	Subtype:Luminal A | Stage:NA | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M028.mzML
SUBJECT_SAMPLE_FACTORS           	-	M029	Subtype:Luminal A | Stage:I | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M029.mzML
SUBJECT_SAMPLE_FACTORS           	-	M032	Subtype:Luminal B | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M032.mzML
SUBJECT_SAMPLE_FACTORS           	-	M033	Subtype:HER2 | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M033.mzML
SUBJECT_SAMPLE_FACTORS           	-	M034	Subtype:Luminal B | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M034.mzML
SUBJECT_SAMPLE_FACTORS           	-	M037	Subtype:Basal like | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M037.mzML
SUBJECT_SAMPLE_FACTORS           	-	M038	Subtype:Basal like | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M038.mzML
SUBJECT_SAMPLE_FACTORS           	-	M039	Subtype:Luminal B | Stage:IV | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M039.mzML
SUBJECT_SAMPLE_FACTORS           	-	M040	Subtype:Luminal B | Stage:I | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M040.mzML
SUBJECT_SAMPLE_FACTORS           	-	M041	Subtype:Luminal A | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M041.mzML
SUBJECT_SAMPLE_FACTORS           	-	M042	Subtype:Luminal A | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M042.mzML
SUBJECT_SAMPLE_FACTORS           	-	M045	Subtype:Basal like | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M045.mzML
SUBJECT_SAMPLE_FACTORS           	-	M046	Subtype:HER2 | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M046.mzML
SUBJECT_SAMPLE_FACTORS           	-	M047	Subtype:Luminal A | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M047.mzML
SUBJECT_SAMPLE_FACTORS           	-	M048	Subtype:Luminal B | Stage:IV | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M048.mzML
SUBJECT_SAMPLE_FACTORS           	-	M052	Subtype:Luminal B | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M052.mzML
SUBJECT_SAMPLE_FACTORS           	-	M055	Subtype:Luminal A | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M055.mzML
SUBJECT_SAMPLE_FACTORS           	-	M060	Subtype:Luminal A | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M060.mzML
SUBJECT_SAMPLE_FACTORS           	-	M064	Subtype:HER2 | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M064.mzML
SUBJECT_SAMPLE_FACTORS           	-	M065	Subtype:Luminal A | Stage:I | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M065.mzML
SUBJECT_SAMPLE_FACTORS           	-	M069	Subtype:Luminal B | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M069.mzML
SUBJECT_SAMPLE_FACTORS           	-	M070	Subtype:Luminal A | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M070.mzML
SUBJECT_SAMPLE_FACTORS           	-	M071	Subtype:Luminal B | Stage:NA | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M071.mzML
SUBJECT_SAMPLE_FACTORS           	-	M076	Subtype:Luminal B | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M076.mzML
SUBJECT_SAMPLE_FACTORS           	-	M081	Subtype:HER2 | Stage:I | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M081.mzML
SUBJECT_SAMPLE_FACTORS           	-	M085	Subtype:Luminal A | Stage:I | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M085.mzML
SUBJECT_SAMPLE_FACTORS           	-	M093	Subtype:Luminal B | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M093.mzML
SUBJECT_SAMPLE_FACTORS           	-	M094	Subtype:Luminal B | Stage:I | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M094.mzML
SUBJECT_SAMPLE_FACTORS           	-	M096	Subtype:Luminal B | Stage:I | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M096.mzML
SUBJECT_SAMPLE_FACTORS           	-	M105	Subtype:HER2 | Stage:IV | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M105.mzML
SUBJECT_SAMPLE_FACTORS           	-	M107	Subtype:Luminal B | Stage:IV | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M107.mzML
SUBJECT_SAMPLE_FACTORS           	-	M110	Subtype:Luminal B | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M110.mzML
SUBJECT_SAMPLE_FACTORS           	-	M112	Subtype:HER2 | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M112.mzML
SUBJECT_SAMPLE_FACTORS           	-	M113	Subtype:Luminal B | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M113.mzML
SUBJECT_SAMPLE_FACTORS           	-	M116	Subtype:Luminal A | Stage:I | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M116.mzML
SUBJECT_SAMPLE_FACTORS           	-	M117	Subtype:Luminal A | Stage:NA | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M117.mzML
SUBJECT_SAMPLE_FACTORS           	-	M118	Subtype:HER2 | Stage:NA | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M118.mzML
SUBJECT_SAMPLE_FACTORS           	-	M120	Subtype:Basal like | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M120.mzML
SUBJECT_SAMPLE_FACTORS           	-	M121	Subtype:HER2 | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M121.mzML
SUBJECT_SAMPLE_FACTORS           	-	M122	Subtype:Luminal A | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M122.mzML
SUBJECT_SAMPLE_FACTORS           	-	M125	Subtype:Luminal B | Stage:NA | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M125.mzML
SUBJECT_SAMPLE_FACTORS           	-	M126	Subtype:HER2 | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M126.mzML
SUBJECT_SAMPLE_FACTORS           	-	M130	Subtype:Luminal A | Stage:I | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M130.mzML
SUBJECT_SAMPLE_FACTORS           	-	M138	Subtype:Control | Stage:Control | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M138.mzML
SUBJECT_SAMPLE_FACTORS           	-	M141	Subtype:Control | Stage:Control | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M141.mzML
SUBJECT_SAMPLE_FACTORS           	-	M145	Subtype:HER2 | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M145.mzML
SUBJECT_SAMPLE_FACTORS           	-	M147	Subtype:Basal like | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M147.mzML
SUBJECT_SAMPLE_FACTORS           	-	M148	Subtype:Control | Stage:Control | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M148.mzML
SUBJECT_SAMPLE_FACTORS           	-	M149	Subtype:Control | Stage:Control | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M149.mzML
SUBJECT_SAMPLE_FACTORS           	-	M151	Subtype:Basal like | Stage:NA | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M151.mzML
SUBJECT_SAMPLE_FACTORS           	-	M152	Subtype:Basal like | Stage:NA | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M152.mzML
SUBJECT_SAMPLE_FACTORS           	-	M155	Subtype:Luminal A | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M155.mzML
SUBJECT_SAMPLE_FACTORS           	-	M156	Subtype:Luminal A | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M156.mzML
SUBJECT_SAMPLE_FACTORS           	-	M159	Subtype:Basal like | Stage:NA | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M159.mzML
SUBJECT_SAMPLE_FACTORS           	-	M162	Subtype:HER2 | Stage:III | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M162.mzML
SUBJECT_SAMPLE_FACTORS           	-	M163	Subtype:Control | Stage:Control | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M163.mzML
SUBJECT_SAMPLE_FACTORS           	-	M165	Subtype:HER2 | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M165.mzML
SUBJECT_SAMPLE_FACTORS           	-	M166	Subtype:Control | Stage:Control | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M166.mzML
SUBJECT_SAMPLE_FACTORS           	-	M167	Subtype:Control | Stage:Control | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M167.mzML
SUBJECT_SAMPLE_FACTORS           	-	M170	Subtype:Control | Stage:Control | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M170.mzML
SUBJECT_SAMPLE_FACTORS           	-	M171	Subtype:Control | Stage:Control | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M171.mzML
SUBJECT_SAMPLE_FACTORS           	-	M173	Subtype:Luminal B | Stage:II | Sample source:Plasma	RAW_FILE_NAME(Raw data)=M173.mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	This cross-sectional, exploratory study with an analytical component included
CO:COLLECTION_SUMMARY            	141 cases of women aged 25 to 80 years with newly diagnosed primary breast
CO:COLLECTION_SUMMARY            	cancer, at any stage and biological subtype.
CO:SAMPLE_TYPE                   	Blood (plasma)
#TREATMENT
TR:TREATMENT_SUMMARY             	All patients were treated at Hospital Universitario Mayor Méderi and consented
TR:TREATMENT_SUMMARY             	to the collection of clinical information and blood samples for participation in
TR:TREATMENT_SUMMARY             	the study. Exclusion criteria included previous adjuvant or systemic oncologic
TR:TREATMENT_SUMMARY             	treatment, as well as a history of autoimmune diseases, genetic metabolic
TR:TREATMENT_SUMMARY             	disorders, pharmacologic or HIV-related immunosuppression, and any other type of
TR:TREATMENT_SUMMARY             	cancer or tumor aside from breast cancer. A control group of 14 women without
TR:TREATMENT_SUMMARY             	breast cancer was also included. Absence of malignancy was confirmed by
TR:TREATMENT_SUMMARY             	mammography or breast ultrasound within the past year, with a BIRADS score <2
TR:TREATMENT_SUMMARY             	and no medical history of any type of cancer. All control participants were
TR:TREATMENT_SUMMARY             	alive and cancer-free at the time of sample collection. These control samples
TR:TREATMENT_SUMMARY             	were included primarily to serve as a quality control step and to help preserve
TR:TREATMENT_SUMMARY             	the analytical stability of the models. This study was approved by the
TR:TREATMENT_SUMMARY             	institutional ethics committees of Universidad del Rosario and Hospital
TR:TREATMENT_SUMMARY             	Universitario Mayor Méderi. Participants completed a questionnaire administered
TR:TREATMENT_SUMMARY             	in collaboration with co-investigators, which collected data on the most
TR:TREATMENT_SUMMARY             	prevalent multifactorial and metabolic comorbidities observed among Colombian
TR:TREATMENT_SUMMARY             	women, particularly those with breast cancer, such as dyslipidemia, primary
TR:TREATMENT_SUMMARY             	hypothyroidism of non-autoimmune or idiopathic origin, type II diabetes
TR:TREATMENT_SUMMARY             	mellitus, obesity, and arterial hypertension. Participants’ current height and
TR:TREATMENT_SUMMARY             	weight were also recorded. Body mass index (BMI) was calculated as weight in
TR:TREATMENT_SUMMARY             	kilograms divided by height in meters squared. The questionnaire additionally
TR:TREATMENT_SUMMARY             	gathered information on age and hormonal status.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Sample extraction was conducted by taking 50 µL of plasma sample, and 150 µL
SP:SAMPLEPREP_SUMMARY            	of MeOH (at -20°C) was added and vortexed at 3200 rpm for 3 minutes. The
SP:SAMPLEPREP_SUMMARY            	samples were incubated at -20 °C for 20 minutes and centrifuged at 13000 rpm,
SP:SAMPLEPREP_SUMMARY            	4°C for 10 minutes
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	The metabolomic analysis was performed using an LC-QTOF-MS Agilent 6545
CH:CHROMATOGRAPHY_SUMMARY        	instru-ment. Chromatographic separation was carried out using a C18 column
CH:CHROMATOGRAPHY_SUMMARY        	(InfinityLab Po-roshell 120 EC-C18 100 x 2.1 mm, 1.9 µm) at 40 °C and a
CH:CHROMATOGRAPHY_SUMMARY        	gradient elution composed of: 0.1 % (v/v) formic acid in Milli-Q water (Phase A)
CH:CHROMATOGRAPHY_SUMMARY        	and 0.1 % (v/v) formic acid in acetonitrile (Phase B) with a constant flow of
CH:CHROMATOGRAPHY_SUMMARY        	0.4 mL/min. The elution gradient started with 5% phase B, increasing to 95% B
CH:CHROMATOGRAPHY_SUMMARY        	during the first 15 minutes, and was kept constant for 1 minute. Finally, the
CH:CHROMATOGRAPHY_SUMMARY        	percentage for phase B was reduced to 5% from minutes 16-18, remaining at the
CH:CHROMATOGRAPHY_SUMMARY        	initial conditions for 4 minutes for a final analysis time of 22 minutes. Mass
CH:CHROMATOGRAPHY_SUMMARY        	spectrom-etry detection was performed in positive ionization mode at scan range
CH:CHROMATOGRAPHY_SUMMARY        	from 50 to 1100 m/z.
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent 6545
CH:COLUMN_NAME                   	InfinityLab Poroshell 120 EC-C18 (100 x 2.1mm, 1.9um)
CH:SOLVENT_A                     	100% water; 0.1% formic acid
CH:SOLVENT_B                     	100% acetonitrile; 0.1% formic acid
CH:FLOW_GRADIENT                 	The elution gradient started with 5% phase B, increasing to 95% B during the
CH:FLOW_GRADIENT                 	first 15 minutes, and was kept constant for 1 minute. Finally, the percentage
CH:FLOW_GRADIENT                 	for phase B was re-duced to 5% from minutes 16-18, remaining at the initial
CH:FLOW_GRADIENT                 	conditions for 4 minutes for a fi-nal analysis time of 22 minutes
CH:FLOW_RATE                     	0.4 mL/min
CH:COLUMN_TEMPERATURE            	30
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Agilent 6545 QTOF
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	The raw data by LC-QTOF-MS were processed using Agilent MassHunter Profinder
MS:MS_COMMENTS                   	10.0 software to perform deconvolution, alignment, and integration. For
MS:MS_COMMENTS                   	GC-MS-QTOF data were processed in the Agilent MassHunter Unknowns Analysis,
MS:MS_COMMENTS                   	Agilent Mass Pro-filer Professional, and Agilent MassHunter Quantitative
MS:MS_COMMENTS                   	software. These programs will be used to perform deconvolution, alignment,
MS:MS_COMMENTS                   	integration, and identification of the metabo-lites present in the samples.
MS:MS_RESULTS_FILE               	ST003936_AN006478_Results.txt	UNITS:Area	Has m/z:Neutral masses	Has RT:Yes	RT units:Minutes
#END