#METABOLOMICS WORKBENCH siriphan_man_20250605_195307 DATATRACK_ID:5998 STUDY_ID:ST004177 ANALYSIS_ID:AN006932 PROJECT_ID:PR002474
VERSION             	1
CREATED_ON             	June 6, 2025, 2:30 am
#PROJECT
PR:PROJECT_TITLE                 	Plasma Metabolomic Analysis in Thai EGFR-mutated Non-Small Cell Lung Cancer
PR:PROJECT_TITLE                 	Patients
PR:PROJECT_SUMMARY               	Lung cancer remains the leading cause of cancer-related deaths worldwide,
PR:PROJECT_SUMMARY               	emphasizing an urgent need for effective, non-invasive diagnostic and prognostic
PR:PROJECT_SUMMARY               	biomarkers. Metabolic reprogramming driven by oncogenic driver alterations,
PR:PROJECT_SUMMARY               	especially EGFR mutations in non-small cell lung cancer (NSCLC), provides
PR:PROJECT_SUMMARY               	promising metabolic signatures detectable in plasma. This study aimed to
PR:PROJECT_SUMMARY               	identify plasma metabolomic biomarkers distinguishing healthy individuals from
PR:PROJECT_SUMMARY               	NSCLC patients and to further stratify NSCLC patients by EGFR mutation status
PR:PROJECT_SUMMARY               	and EGFR tyrosine kinase inhibitor (TKI) resistance. Using gas
PR:PROJECT_SUMMARY               	chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass
PR:PROJECT_SUMMARY               	spectrometry (LC-MS/MS), we quantified four groups of targeted metabolites
PR:PROJECT_SUMMARY               	(amino acids, organic acids, sugar and sugar alcohols, and phospholipids) of 78
PR:PROJECT_SUMMARY               	individuals, including NSCLC patients and healthy controls. Metabolomic
PR:PROJECT_SUMMARY               	signatures distinctly separated healthy controls from NSCLC patients and,
PR:PROJECT_SUMMARY               	furthermore, EGFR-mutated NSCLC from EGFR wild-type cases. Additionally,
PR:PROJECT_SUMMARY               	metabolomics results effectively distinguished EGFR TKI-resistant from
PR:PROJECT_SUMMARY               	treatment-naïve EGFR-mutated NSCLC. Our findings highlight the potential of
PR:PROJECT_SUMMARY               	plasma metabolites as minimally invasive biomarkers for molecular classification
PR:PROJECT_SUMMARY               	and monitoring therapeutic response in NSCLC patients.
PR:INSTITUTE                     	Mahidol University
PR:LAST_NAME                     	Khoomrung
PR:FIRST_NAME                    	Sakda
PR:ADDRESS                       	2 Wanglang road
PR:EMAIL                         	Sakda.kho@mahidol.edu
PR:PHONE                         	024195505
#STUDY
ST:STUDY_TITLE                   	Plasma Organic acids Analysis in Thai EGFR-mutated Non-Small Cell Lung Cancer
ST:STUDY_TITLE                   	Patients
ST:STUDY_SUMMARY                 	Lung cancer remains the leading cause of cancer-related deaths worldwide,
ST:STUDY_SUMMARY                 	emphasizing an urgent need for effective, non-invasive diagnostic and prognostic
ST:STUDY_SUMMARY                 	biomarkers. Metabolic reprogramming driven by oncogenic driver alterations,
ST:STUDY_SUMMARY                 	especially EGFR mutations in non-small cell lung cancer (NSCLC), provides
ST:STUDY_SUMMARY                 	promising metabolic signatures detectable in plasma. This study aimed to
ST:STUDY_SUMMARY                 	identify plasma metabolomic biomarkers distinguishing healthy individuals from
ST:STUDY_SUMMARY                 	NSCLC patients and to further stratify NSCLC patients by EGFR mutation status
ST:STUDY_SUMMARY                 	and EGFR tyrosine kinase inhibitor (TKI) resistance. Using gas
ST:STUDY_SUMMARY                 	chromatography-mass spectrometry (GC-MS), we quantified 11 organic acids of 78
ST:STUDY_SUMMARY                 	individuals, including NSCLC patients and healthy controls.
ST:INSTITUTE                     	Mahidol University
ST:LAST_NAME                     	Khoomrung
ST:FIRST_NAME                    	Sakda
ST:ADDRESS                       	2 Wanglang road
ST:EMAIL                         	siriphan.man@mahidol.edu
ST:PHONE                         	025195505
#SUBJECT
SU:SUBJECT_TYPE                  	Human
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
SU:GENDER                        	Male and 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           	-	A01	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A01.D
SUBJECT_SAMPLE_FACTORS           	-	A02	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A02.D
SUBJECT_SAMPLE_FACTORS           	-	A13	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A13.D
SUBJECT_SAMPLE_FACTORS           	-	A14	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A14.D
SUBJECT_SAMPLE_FACTORS           	-	A20	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A20.D
SUBJECT_SAMPLE_FACTORS           	-	A22	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A22.D
SUBJECT_SAMPLE_FACTORS           	-	A23	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A23.D
SUBJECT_SAMPLE_FACTORS           	-	A24	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A24.D
SUBJECT_SAMPLE_FACTORS           	-	A25	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A25.D
SUBJECT_SAMPLE_FACTORS           	-	A26	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A26.D
SUBJECT_SAMPLE_FACTORS           	-	A27	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A27.D
SUBJECT_SAMPLE_FACTORS           	-	A28	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A28.D
SUBJECT_SAMPLE_FACTORS           	-	A29	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A29.D
SUBJECT_SAMPLE_FACTORS           	-	A30	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A30.D
SUBJECT_SAMPLE_FACTORS           	-	A31	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A31.D
SUBJECT_SAMPLE_FACTORS           	-	A32	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A32.D
SUBJECT_SAMPLE_FACTORS           	-	A33	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A33.D
SUBJECT_SAMPLE_FACTORS           	-	A34	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A34.D
SUBJECT_SAMPLE_FACTORS           	-	A35	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A35-2.D
SUBJECT_SAMPLE_FACTORS           	-	A36	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A36.D
SUBJECT_SAMPLE_FACTORS           	-	A37	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A37.D
SUBJECT_SAMPLE_FACTORS           	-	A38	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A38.D
SUBJECT_SAMPLE_FACTORS           	-	A39	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A39.D
SUBJECT_SAMPLE_FACTORS           	-	A40	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A40.D
SUBJECT_SAMPLE_FACTORS           	-	A41	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A41.D
SUBJECT_SAMPLE_FACTORS           	-	A42	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A42.D
SUBJECT_SAMPLE_FACTORS           	-	A43	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A43.D
SUBJECT_SAMPLE_FACTORS           	-	A45	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A45.D
SUBJECT_SAMPLE_FACTORS           	-	A46	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=A46.D
SUBJECT_SAMPLE_FACTORS           	-	B01	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B01.D
SUBJECT_SAMPLE_FACTORS           	-	B02	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B02.D
SUBJECT_SAMPLE_FACTORS           	-	B05	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B05.D
SUBJECT_SAMPLE_FACTORS           	-	B06	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B06.D
SUBJECT_SAMPLE_FACTORS           	-	B07	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B07.D
SUBJECT_SAMPLE_FACTORS           	-	B08	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B08.D
SUBJECT_SAMPLE_FACTORS           	-	B09	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B09.D
SUBJECT_SAMPLE_FACTORS           	-	B10	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B10.D
SUBJECT_SAMPLE_FACTORS           	-	B11	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B11.D
SUBJECT_SAMPLE_FACTORS           	-	B12	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B12.D
SUBJECT_SAMPLE_FACTORS           	-	B13	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B13.D
SUBJECT_SAMPLE_FACTORS           	-	B14	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B14.D
SUBJECT_SAMPLE_FACTORS           	-	B15	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B15.D
SUBJECT_SAMPLE_FACTORS           	-	B16	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B16.D
SUBJECT_SAMPLE_FACTORS           	-	B17	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B17.D
SUBJECT_SAMPLE_FACTORS           	-	B18	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B18.D
SUBJECT_SAMPLE_FACTORS           	-	B19	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B19.D
SUBJECT_SAMPLE_FACTORS           	-	B21	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B21.D
SUBJECT_SAMPLE_FACTORS           	-	B22	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B22.D
SUBJECT_SAMPLE_FACTORS           	-	B23	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B23.D
SUBJECT_SAMPLE_FACTORS           	-	B24	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B24.D
SUBJECT_SAMPLE_FACTORS           	-	B25	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B25.D
SUBJECT_SAMPLE_FACTORS           	-	B26	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B26.D
SUBJECT_SAMPLE_FACTORS           	-	B27	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B27.D
SUBJECT_SAMPLE_FACTORS           	-	B28	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B28.D
SUBJECT_SAMPLE_FACTORS           	-	B30	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B30.D
SUBJECT_SAMPLE_FACTORS           	-	B33	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B33.D
SUBJECT_SAMPLE_FACTORS           	-	B34	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B34.D
SUBJECT_SAMPLE_FACTORS           	-	B42	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	RAW_FILE_NAME(Organic acids RAW file)=B42.D
SUBJECT_SAMPLE_FACTORS           	-	C01	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	RAW_FILE_NAME(Organic acids RAW file)=C01.D
SUBJECT_SAMPLE_FACTORS           	-	C02	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	RAW_FILE_NAME(Organic acids RAW file)=C02.D
SUBJECT_SAMPLE_FACTORS           	-	C03	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	RAW_FILE_NAME(Organic acids RAW file)=C03.D
SUBJECT_SAMPLE_FACTORS           	-	C04	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	RAW_FILE_NAME(Organic acids RAW file)=C04.D
SUBJECT_SAMPLE_FACTORS           	-	C05	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	RAW_FILE_NAME(Organic acids RAW file)=C05.D
SUBJECT_SAMPLE_FACTORS           	-	C06	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	RAW_FILE_NAME(Organic acids RAW file)=C06.D
SUBJECT_SAMPLE_FACTORS           	-	C07	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	RAW_FILE_NAME(Organic acids RAW file)=C07.D
SUBJECT_SAMPLE_FACTORS           	-	C08	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	RAW_FILE_NAME(Organic acids RAW file)=C08.D
SUBJECT_SAMPLE_FACTORS           	-	C09	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	RAW_FILE_NAME(Organic acids RAW file)=C09.D
SUBJECT_SAMPLE_FACTORS           	-	C10	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	RAW_FILE_NAME(Organic acids RAW file)=C10.D
SUBJECT_SAMPLE_FACTORS           	-	D01	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	RAW_FILE_NAME(Organic acids RAW file)=D01.D
SUBJECT_SAMPLE_FACTORS           	-	D02	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	RAW_FILE_NAME(Organic acids RAW file)=D02.D
SUBJECT_SAMPLE_FACTORS           	-	D03	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	RAW_FILE_NAME(Organic acids RAW file)=D03.D
SUBJECT_SAMPLE_FACTORS           	-	D04	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	RAW_FILE_NAME(Organic acids RAW file)=D04.D
SUBJECT_SAMPLE_FACTORS           	-	D05	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	RAW_FILE_NAME(Organic acids RAW file)=D05.D
SUBJECT_SAMPLE_FACTORS           	-	D06	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	RAW_FILE_NAME(Organic acids RAW file)=D06.D
SUBJECT_SAMPLE_FACTORS           	-	D08	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	RAW_FILE_NAME(Organic acids RAW file)=D08.D
SUBJECT_SAMPLE_FACTORS           	-	D09	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	RAW_FILE_NAME(Organic acids RAW file)=D09.D
SUBJECT_SAMPLE_FACTORS           	-	D10	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	RAW_FILE_NAME(Organic acids RAW file)=D10.D
SUBJECT_SAMPLE_FACTORS           	-	D11	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	RAW_FILE_NAME(Organic acids RAW file)=D11.D
#COLLECTION
CO:COLLECTION_SUMMARY            	Blood was drawn directly into Cell-Free DNA BCT® tubes to stabilize cell-free
CO:COLLECTION_SUMMARY            	DNA. Plasma was separated from cellular components by centrifugation. For NSCLC
CO:COLLECTION_SUMMARY            	patients, plasma was first subjected to EGFR mutation analysis. Plasma aliquots
CO:COLLECTION_SUMMARY            	from all participants were stored at −80°C until subsequent analysis.
CO:SAMPLE_TYPE                   	Blood (plasma)
#TREATMENT
TR:TREATMENT_SUMMARY             	Treatment-naïve NSCLC patients had never received any systemic treatment for
TR:TREATMENT_SUMMARY             	lung cancer. EGFR TKI-resistant NSCLC patients had evidence of disease
TR:TREATMENT_SUMMARY             	progression according to the Response Evaluation Criteria In Solid Tumors
TR:TREATMENT_SUMMARY             	(RECIST) version 1.1 after receiving first- or second-generation EGFR TKI
TR:TREATMENT_SUMMARY             	treatment. All EGFR TKI-resistant NSCLC patients had acquired EGFR T790M
TR:TREATMENT_SUMMARY             	mutation.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	For organic acids, plasma samples (50 µL) was diluted with 75 µL of ultra-pure
SP:SAMPLEPREP_SUMMARY            	water (Type I) in a 4 mL HPLC vial. Then, 50 µL of trans-cinnamic acid (0.5 mM)
SP:SAMPLEPREP_SUMMARY            	was added into the vial as an internal standard, followed by 400 µl of diethyl
SP:SAMPLEPREP_SUMMARY            	ether to extract organic acid from samples. The vial containing the mixture was
SP:SAMPLEPREP_SUMMARY            	vortexed for 2 min. After that, the mixture was centrifuged at 1400 rpm for 5
SP:SAMPLEPREP_SUMMARY            	min using a concentrator. The 100 µL of the upper layer was collected and
SP:SAMPLEPREP_SUMMARY            	transferred into an autosampler crimp top vial using a positive displacement
SP:SAMPLEPREP_SUMMARY            	micropipette. After the organic layer was transferred, 50 µL of MTBSTFA was
SP:SAMPLEPREP_SUMMARY            	added as the derivatizing agent. The autosampler vial was sealed with an
SP:SAMPLEPREP_SUMMARY            	aluminum crimp cap with PTFE/Silicone septa. The vial was then incubated at 80
SP:SAMPLEPREP_SUMMARY            	ºC for 40 min. After incubation, samples were analyzed using GC-MS.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	The chromatographic analysis was performed on a gas chromatography (Agilent;
CH:CHROMATOGRAPHY_SUMMARY        	7890B) equipped with a mass spectrometer (Agilent, 7000D) and PAL auto sampler
CH:CHROMATOGRAPHY_SUMMARY        	system. Aliquots of the derivatized amino acids (1 µl) were injected by using
CH:CHROMATOGRAPHY_SUMMARY        	split mode at 1:20 split ratio at 280°C into a DB-1MS column (30 m x 0.25um x
CH:CHROMATOGRAPHY_SUMMARY        	0.25 mm i.d.). Helium was used as a carrier gas with a constant flow rate of 1.1
CH:CHROMATOGRAPHY_SUMMARY        	ml/min. The initial oven temperature was controlled at 80°C, held for 3 min.
CH:CHROMATOGRAPHY_SUMMARY        	The GC oven was programmed as follows: ramp from 80°C to 160°C (50 °C/min),
CH:CHROMATOGRAPHY_SUMMARY        	held for 3 min and ramp from 160°C to 340°C (80 °C/min). The transfer line,
CH:CHROMATOGRAPHY_SUMMARY        	ion source (EI), and quadrupole were set as 325 °C, 240 °C, and 180 °C,
CH:CHROMATOGRAPHY_SUMMARY        	respectively
CH:CHROMATOGRAPHY_TYPE           	GC
CH:INSTRUMENT_NAME               	Agilent 7890B
CH:COLUMN_NAME                   	DB-1MS column (30 m x 0.25um x 0.25 mm)
CH:SOLVENT_A                     	not applicable
CH:SOLVENT_B                     	not applicable
CH:FLOW_GRADIENT                 	not applicable
CH:FLOW_RATE                     	1.1ml/min
CH:COLUMN_TEMPERATURE            	not applicable
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Agilent 7000B
MS:INSTRUMENT_TYPE               	Triple quadrupole
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	The mass spectrometer was operated in Multiple Reaction Monitoring (MRM) mode.
MS:MS_COMMENTS                   	MS data was acquired using MassHunter software (version 10.0). The calibration
MS:MS_COMMENTS                   	curves were performed using the mixture of volatile (formic acid, acetic acid,
MS:MS_COMMENTS                   	propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid,
MS:MS_COMMENTS                   	isocaproic acid, hexanoic acid and heptanoic acid) and non-volatile (lactic
MS:MS_COMMENTS                   	acid, pyruvic acid, oxalic acid, malonic acid, methyl malonic acid, succinic
MS:MS_COMMENTS                   	acid, fumaric acid, malic acid) organic acids at different concentrations ranged
MS:MS_COMMENTS                   	from 0.78 ppm to 25 ppm. Qualitative and quantitative analyses were performed on
MS:MS_COMMENTS                   	Agilent MassHunter software (version 10.0 Agilent Technologies, USA), and
MS:MS_COMMENTS                   	exported into Microsoft Excel for further data processing.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	micro molar
MS_METABOLITE_DATA_START
Samples	A01	A02	A13	A14	A20	A22	A23	A24	A25	A26	A27	A28	A29	A30	A31	A32	A33	A34	A35	A36	A37	A38	A39	A40	A41	A42	A43	A45	A46	B01	B02	B05	B06	B07	B08	B09	B10	B11	B12	B13	B14	B15	B16	B17	B18	B19	B21	B22	B23	B24	B25	B26	B27	B28	B30	B33	B34	B42	C01	C02	C03	C04	C05	C06	C07	C08	C09	C10	D01	D02	D03	D04	D05	D06	D08	D09	D10	D11
Factors	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:NSCLC | EGFR mutation:No | Treatment:Naïve	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	Sample source:Plasma | Disease:Healthy | EGFR mutation:n/a | Treatment:n/a	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant	Sample source:Plasma | Disease:NSCLC | EGFR mutation:Yes | Treatment:TKI resistant
Formic acid	204.748325	293.6004	260.407825	255.731975	360.32915	216.2642	176.53595	188.790375	257.51235	192.221575	383.2485	122.1537	186.477825	203.5542	226.770625	380.760525	158.222675	269.99705	89.849	293.717075	119.43215	410.084025	111.08695	243.543575	117.60045	73.279925	153.4618	163.21595	176.471875	184.740875	257.804	232.3234	178.762625	141.9072	173.5311	283.95565	37.5967	96.65675	213.789975	89.795625	183.471575	295.880325	170.192075	161.462175	96.3183	52.942175	228.7614	181.712375	121.362075	132.336275	145.5214	188.6972	106.889725	89.442325	153.692325	72.81305	215.43085	90.551925	15.855875	28.934825	76.1706	28.502775	96.5089	121.161325	17.91355	8.5403	55.685525	10.341275	95.13095	89.663775	170.0664	220.272775	178.822275	147.98015	149.456525	45.477925	209.940125	416.867275
Acetic acid	30.83515	36.6333	33.328375	26.299775	29.988225	30.89285	29.890275	29.053175	28.09595	33.0284	46.297725	27.7499	30.275775	32.9858	33.00745	29.04665	31.97015	28.71985	26.0044	36.1816	24.933925	28.8201	28.25935	24.5615	29.2699	24.307075	29.4131	29.0792	33.056625	68.588875	33.813175	31.4203	31.2349	29.2484	34.56875	31.4459	24.97865	27.99095	37.755325	24.97655	25.98685	37.1487	25.489375	31.2709	29.5784	30.834275	36.23095	24.653175	31.164475	30.4075	28.800825	35.311325	29.417475	32.16735	27.88085	27.2511	29.27935	31.314175	35.32725	36.964575	42.324575	35.79365	46.113325	44.712325	32.38695	34.4606	37.1796	31.314275	24.805325	28.065325	25.989425	28.9971	26.44945	22.6969	32.753575	27.808	31.977375	34.415225
Propionic acid	15.739875	15.854575	15.729325	12.54435	13.245	13.67125	14.432375	14.598825	13.4899	14.024525	14.9336	14.9668	14.6492	14.009775	13.065475	12.73185	14.4099	13.294925	12.654625	15.954	14.1034	13.944925	13.5459	11.30575	13.412175	14.107175	12.8552	13.416375	14.38615	13.555275	13.645875	14.77275	15.180375	14.274125	13.26275	13.6301	10.739375	14.4044	13.471925	12.511125	14.252075	14.31995	13.2902	13.31575	15.065575	14.231925	14.47845	12.758375	13.5933	14.187	13.605575	14.973375	14.207225	13.596925	14.174075	13.6907	13.971025	14.02855	46.75085	47.47395	44.05655	44.5975	44.501	49.0207	44.78835	46.733475	44.3053	47.768375	12.72165	13.294025	12.01835	12.8426	13.869925	12.17705	13.627675	13.929975	14.091625	13.0747
Isobutyric acid	0.88065	0.925325	0.870375	0.63845	0.58265	0.853775	0.871375	0.8054	0.7718	0.80825	0.711	0.671725	0.67565	0.628125	0.5605	0.387075	0.478225	0.5694	0.523825	0.6996	0.5704	0.7434	0.82465	0.6856	0.638775	0.602175	0.6813	0.46065	0.717075	1.27905	0.63445	0.621025	0.52	0.805775	0.52425	0.5733	0.7237	0.610025	0.6182	0.674125	0.562025	0.5625	0.61235	0.852975	0.834	1.081675	0.73525	0.6309	0.739825	0.747075	0.59625	0.6334	0.772775	0.741075	0.751875	0.614425	0.668775	0.601725	0.633425	0.573225	0.522125	0.5778	0.65095	0.6009	0.689175	0.6454	0.476475	0.661775	0.69185	0.63685	0.58925	0.566075	0.702	0.715375	0.79385	0.667075	0.66585	0.585975
Butyric acid	0.552275	0.390525	0.359525	0.44765	0.4038	0.5192	0.6445	0.668025	0.521275	0.646975	0.35795	0.67035	0.557525	0.526725	0.434725	0.31705	0.335275	0.50445	0.31845	0.41375	0.431425	0.451475	0.5064	0.3261	0.38395	0.446175	0.5059	0.467975	0.45015	0.4797	0.497225	0.45185	0.47865	0.451575	0.490075	0.5059	0.47685	0.598	0.552425	0.336125	0.365775	0.450875	0.506725	0.376925	0.896525	0.50915	0.4309	0.466825	0.4314	0.437875	0.50505	0.409125	0.40215	0.4587	0.4565	0.37085	0.3939	0.48685	0.552475	0.416675	0.4719	0.57045	0.489275	0.42375	0.643825	0.636825	0.513975	0.5393	0.42425	0.450125	0.562675	0.3747	0.434125	0.338575	0.3328	0.4057	0.399575	0.476725
Hexanoic acid	0.427075	0.36855	0.539675	0.468375	0.385275	0.39735	0.31885	0.48705	0.3233	0.574725	0.4613	0.26935	0.407825	0.188075	0.1582	0.141075	0.2021	0.162875	0.161575	0.22085	0.161	0.250475	0.176	0.164825	0.13565	0.145825	0.304475	0.19365	0.217975	0.48365	0.362375	0.13125	0.177725	0.29345	0.11605	0.100775	0.157125	0.2629	0.127825	0.10315	0.15785	0.210275	0.19425	0.298525	0.1826	0.246575	0.2719	0.291275	0.23695	0.1716	0.249225	0.2357	0.202075	0.1443	0.238675	0.18965	0.176075	0.121025	0.23505	0.11905	0.156875	0.199575	0.1483	0.2372	0.18215	0.1796	0.116725	0.23755	0.218275	0.201425	0.236875	0.219575	0.207625	0.2695	0.271775	0.21765	0.257475	0.353325
Lactic acid	32.8698	42.844075	64.010825	61.605575	106.390375	60.74315	59.7423	41.341925	74.770375	28.28725	51.59585	36.25545	64.0582	37.124025	25.2678	46.208875	9.56285	100.262775	34.759875	70.2473	38.857775	113.843625	29.27565	80.8211	24.830325	21.94205	48.337575	70.326075	25.887375	18.1499	69.8892	64.6694	43.57365	33.887875	17.372675	54.698525	13.278175	26.231475	19.51755	19.5476	53.70585	49.8016	52.7911	58.267125	32.56295	42.956725	58.715775	54.695725	34.4317	44.794425	53.379475	20.396975	25.5948	26.73965	54.31145	21.6668	68.14275	23.735375	56.090825	55.67965	52.52615	52.84985	40.47195	58.13975	53.4005	53.34775	47.6151	46.854125	44.033175	17.7414	62.200375	80.316475	47.34245	56.6406	53.96585	17.562425	52.93935	76.71765
Oxalic acid	0.056679641	10.11312935	5.992748697	4.577299653	9.362436703	6.365020123	9.249983777	8.681859939	4.041824886	2.644985129	4.053789	10.05559952	4.779324739	7.538021982	8.717195404	4.365979811	4.538790101	11.33455564	6.386027393	5.901571773	8.298055153	5.039816156	6.280950927	7.754890985	7.174636787	7.217408581	8.036782146	6.192149272	8.506166624	7.344464534	8.251157571	6.78936803	7.872742202	5.301104053	5.968219548	6.875026967	8.313941695	6.459835221	1.390586139	9.715475918	6.423212333	9.486356693	6.515843894	6.876890801	8.860517817	9.139868899	5.649998694	9.045313229	8.390237605	7.532908762	11.33453481	7.689847296	7.279587235	6.452367309	6.108831143	6.697433146	8.732402492	8.014821854	1.59173226	0.676956457	0.635693074	0.569656433	1.180260261	0.166792848	0.666711851	0.68229728	0.300288244	0.523558359	8.828929159	6.01250022	7.8801764	1.1041	1.089675	0.73045	0.85075	0.633325	0.87605	0.7973
Succinic acid	0.134471657	0.214971901	0.498016407	0.141624932	0.08667668	0.22830201	0.290921815	0.239498024	0.055691917	0.071816214	0.335928419	0.082782784	0.112520624	0.055751362	0.15883803	0.173805087	0.125340317	0.018878639	0.113084801	0.129355251	0.406193508	0.330551792	0.2811415	0.360725109	0.237864025	0.313160804	0.214074821	0.271669313	0.238395274	0.171634678	0.219260745	0.217941623	0.185041761	0.021746761	0.033246171	0.090454339	0.036038937	0.072480499	0.05254514	0.004098144	0.026621831	0.351632967	0.181894332	0.110508271	0.154550992	0.273891778	0.196446003	0.221517731	0.268830128	0.08778775	0.130169408	0.433493648	0.328900432	0.298090904	0.111457652	0.236335747	0.134821857	0.381071909	0.282017714	0.47944074	0.440602873	0.276291243	0.345488172	0.300106067	0.506736042	0.659851177	0.338804692	0.510502371	0.268286612	0.172262188	0.081036272	0.02105	0.053325	0.0469	0.0321	0.02345	0.040525	0.051575
Fumaric acid	0.006875	0.02745	0.01985	0.020325	0.050125	0.01965	0.0298	0.019175	0.0116	0.01595	0.03015	0.024	0.008625	0.0192	0.01025	0.014825	0.002025	0.045825	0.015675	0.022875	0.030475	0.0314	0.025525	0.008875	0.00545	0.007675	0.0314	0.02125	0.0098	0.003875	0.01115	0.03	0.017625	0.019825	0.008125	0.031225	0.00155	0.024875	0.02665	0.02465	0.009525	0.012075	0.0141	0.00855	0.015075	0.01425	0.014675	0.02385	0.0155	0.004425	0.011725	0.0067	0.021725	0.019825	0.05025	0.004425	0.0156	0.011425	0.033025	0.01545	0.0151	0.015525	0.002675	0.012225	0.0163	0.0062	0.01005	0.007825	0.02015	0.0043	0.033175	0.036475	0.012775	0.037975	0.02805	0.02	0.02025	0.0609
Pyruvic acid	LoQ	2.1161	1.50295	0.957125	2.2051	1.512375	1.900025	2.231875	0.98945	0.653475	1.0922	2.433175	1.34375	1.78065	LoQ	LoQ	LoQ	2.32435	1.36	1.4327	1.739375	1.676775	1.530425	1.7596	1.631775	1.931875	1.650475	1.49455	1.821475	1.86685	1.522	1.119475	1.701825	1.399825	1.715775	1.60035	1.776	1.472325	0.3004	2.232225	1.649475	2.142925	1.347425	1.760475	2.2349	2.12335	1.665125	2.0757	1.89845	1.409075	2.6334	1.924	1.7232	1.311475	1.668925	1.42915	2.106225	2.061825	0.2952	0.220725	0.29615	0.154775	0.231825	0.190325	0.135925	0.213225	0.24395	0.156125	2.04165	1.42995	1.790925	2.129775	2.079025	1.39685	1.52845	1.2734	1.819025	1.667
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Pubchem CID
Formic acid	284
Acetic acid	176
Propionic acid	1032
Isobutyric acid	6590
Butyric acid	264
Hexanoic acid	8892
Lactic acid	612
Oxalic acid	971
Succinic acid	1110
Fumaric acid	444972
Pyruvic acid	1060
METABOLITES_END
#END