#METABOLOMICS WORKBENCH nurwahidahamdan01_20210916_182707 DATATRACK_ID:2847 STUDY_ID:ST001982 ANALYSIS_ID:AN003233 PROJECT_ID:PR001258
VERSION             	1
CREATED_ON             	September 16, 2021, 8:39 pm
#PROJECT
PR:PROJECT_TITLE                 	Lipidomic characterization of Candida albicans in response to Aureobasidin
PR:PROJECT_TITLE                 	treatment in vitro.
PR:PROJECT_SUMMARY               	Candida albicans is an opportunistic yeast pathogen that causes a wide range of
PR:PROJECT_SUMMARY               	infections especially amongst immunocompromised patients. Aureobasidin A (AbA)
PR:PROJECT_SUMMARY               	has been shown to inhibit inositolphosphoryl ceramide synthase (IPCS), a key
PR:PROJECT_SUMMARY               	enzyme responsible for sphingolipid biosynthesis. There are limited studies
PR:PROJECT_SUMMARY               	exploring IPCS as a target molecule for antifungal treatment. It is hypothesized
PR:PROJECT_SUMMARY               	that the mechanism of AbA inhibition involves alteration of C. albicans
PR:PROJECT_SUMMARY               	phospholipid and sphingolipid profiles. The profiling of C. albicans
PR:PROJECT_SUMMARY               	phospholipid and sphingolipid upon exposure to 0.5-4 µg/ml of AbA were
PR:PROJECT_SUMMARY               	determined using Liquid chromatography-mass spectrometry (LC-MS).
PR:INSTITUTE                     	University of Malaya
PR:DEPARTMENT                    	Medical Microbiology Department
PR:LABORATORY                    	Lab 3
PR:LAST_NAME                     	Hamdan
PR:FIRST_NAME                    	Nur Wahida
PR:ADDRESS                       	Jalan Profesor Diraja Ungku Aziz, 50603 Kuala Lumpur, Wilayah Persekutuan Kuala
PR:ADDRESS                       	Lumpur, Malaysia
PR:EMAIL                         	nurwahidahamdan@siswa.um.edu.my
PR:PHONE                         	0193354272
PR:FUNDING_SOURCE                	FRGS(FP035-2014A), UM PPP(PG178-2015B)
#STUDY
ST:STUDY_TITLE                   	Lipidomic characterization of Candida albicans in response to Aureobasidin
ST:STUDY_TITLE                   	treatment in vitro.
ST:STUDY_SUMMARY                 	Candida albicans is an opportunistic yeast pathogen that causes a wide range of
ST:STUDY_SUMMARY                 	infections especially amongst immunocompromised patients. Aureobasidin A (AbA)
ST:STUDY_SUMMARY                 	has been shown to inhibit inositolphosphoryl ceramide synthase (IPCS), a key
ST:STUDY_SUMMARY                 	enzyme responsible for sphingolipid biosynthesis. There are limited studies
ST:STUDY_SUMMARY                 	exploring IPCS as a target molecule for antifungal treatment. It is hypothesized
ST:STUDY_SUMMARY                 	that the mechanism of AbA inhibition involves alteration of C. albicans
ST:STUDY_SUMMARY                 	phospholipid and sphingolipid profiles. The profiling of C. albicans
ST:STUDY_SUMMARY                 	phospholipid and sphingolipid upon exposure to 0.5-4 µg/ml of AbA were
ST:STUDY_SUMMARY                 	determined using Liquid chromatography-mass spectrometry (LC-MS).
ST:INSTITUTE                     	University of Malaya
ST:LAST_NAME                     	Hamdan
ST:FIRST_NAME                    	Nur Wahida
ST:ADDRESS                       	Jalan Profesor Diraja Ungku Aziz, 50603 Kuala Lumpur, Wilayah Persekutuan Kuala
ST:ADDRESS                       	Lumpur, Malaysia
ST:EMAIL                         	nurwahidahamdan@siswa.um.edu.my
ST:PHONE                         	0193354272
ST:NUM_GROUPS                    	5
ST:TOTAL_SUBJECTS                	Duplicates
ST:NUM_MALES                     	NA
ST:NUM_FEMALES                   	NA
#SUBJECT
SU:SUBJECT_TYPE                  	Yeast
SU:SUBJECT_SPECIES               	Candida albicans
SU:TAXONOMY_ID                   	5476
SU:GENOTYPE_STRAIN               	SC 5314
#FACTORS
#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           	-	N_0.5_PA	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_0.5_PB	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_0.5_SA	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_0.5_SB	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_1_PA	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_1_PB	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_1_SA	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_1_SB	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_2_PA	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_2_PB	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_2_SA	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_2_SB	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_4_PA	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_4_PB	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_4_SA	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_4_SB	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_DMSO_PA	Treatment:Control | Concentration (ug/ml):- | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_DMSO_PB	Treatment:Control | Concentration (ug/ml):- | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_DMSO_SA	Treatment:Control | Concentration (ug/ml):- | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	N_DMSO_SB	Treatment:Control | Concentration (ug/ml):- | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n0.5_PA	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n0.5_PB	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n0.5_SA	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n0.5_SB	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n1_PA	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n1_PB	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n1_SA	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n1_SB	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n2_PA	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n2_PB	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n2_SA	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n2_SB	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n4_PA	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n4_PB	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n4_SA	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	n4_SB	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	nDMSO_PA	Treatment:Control | Concentration (ug/ml):- | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	nDMSO_PB	Treatment:Control | Concentration (ug/ml):- | Extraction method:Phospholipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	nDMSO_SA	Treatment:Control | Concentration (ug/ml):- | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	nDMSO_SB	Treatment:Control | Concentration (ug/ml):- | Extraction method:Sphingolipids | Ionization mode:Negative	
SUBJECT_SAMPLE_FACTORS           	-	P_0.5_PA	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_0.5_PB	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_0.5_SA	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_0.5_SB	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_1_PA	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_1_PB	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_1_SA	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_1_SB	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_2_PA	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_2_PB	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_2_SA	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_2_SB	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_4_PA	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_4_PB	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_4_SA	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_4_SB	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_DMSO_PA	Treatment:Control | Concentration (ug/ml):- | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_DMSO_PB	Treatment:Control | Concentration (ug/ml):- | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_DMSO_SA	Treatment:Control | Concentration (ug/ml):- | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	P_DMSO_SB	Treatment:Control | Concentration (ug/ml):- | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p0.5_PA	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p0.5_PB	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p0.5_SA	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p0.5_SB	Treatment:AbA-treated | Concentration (ug/ml):0.5 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p1_PA	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p1_PB	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p1_SA	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p1_SB	Treatment:AbA-treated | Concentration (ug/ml):1 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p2_PA	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p2_PB	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p2_SA	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p2_SB	Treatment:AbA-treated | Concentration (ug/ml):2 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p4_PA	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p4_PB	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p4_SA	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	p4_SB	Treatment:AbA-treated | Concentration (ug/ml):4 | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	pDMSO_PA	Treatment:Control | Concentration (ug/ml):- | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	pDMSO_PB	Treatment:Control | Concentration (ug/ml):- | Extraction method:Phospholipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	pDMSO_SA	Treatment:Control | Concentration (ug/ml):- | Extraction method:Sphingolipids | Ionization mode:Positive	
SUBJECT_SAMPLE_FACTORS           	-	pDMSO_SB	Treatment:Control | Concentration (ug/ml):- | Extraction method:Sphingolipids | Ionization mode:Positive	
#COLLECTION
CO:COLLECTION_SUMMARY            	A starter culture was prepared by inoculating two loopfuls of C. albicans yeast
CO:COLLECTION_SUMMARY            	colony in 5 ml of yeast peptone dextrose (YPD) broth and incubated at 37°C for
CO:COLLECTION_SUMMARY            	24 hours. 150 microlitres of the starter culture was then inoculated into 150 ml
CO:COLLECTION_SUMMARY            	fresh YPD broth (10^7 of cells/ml) and let to grow for 6 hours until it reached
CO:COLLECTION_SUMMARY            	10^9 of cells/ml.
CO:SAMPLE_TYPE                   	Yeast cells
CO:STORAGE_CONDITIONS            	-80℃
#TREATMENT
TR:TREATMENT_SUMMARY             	The yeast cells were exposed to different concentration of Aureobasidin A (0.5,
TR:TREATMENT_SUMMARY             	1, 2 and 4 microgram/ml). DMSO-treated yeast culture was used as a control. The
TR:TREATMENT_SUMMARY             	cultures were incubated for another 3 hours prior to harvesting. Each of the
TR:TREATMENT_SUMMARY             	conditions was performed in duplicate.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Lipids enrichment Lipids were enriched using a method as described by Guan and
SP:SAMPLEPREP_SUMMARY            	Wenk (2010). After AbA treatment, the yeast cells were harvested and washed
SP:SAMPLEPREP_SUMMARY            	twice. The wet weight was standardized. Briefly, the cells were resuspended in 2
SP:SAMPLEPREP_SUMMARY            	ml 95% ethanol: water: diethyl ether: pyridine: ammonium hydroxide (15 : 15 : 5
SP:SAMPLEPREP_SUMMARY            	: 1 : 0.018). The cells were broken by glass beads (vortexed twice for 1 minute
SP:SAMPLEPREP_SUMMARY            	each) and incubated for 20 min at 60 °C. Debris was pelleted by centrifugation
SP:SAMPLEPREP_SUMMARY            	and the supernatant was transferred to a fresh tube. The pellet was re-extracted
SP:SAMPLEPREP_SUMMARY            	once more using the same procedure. The pooled supernatants were divided into
SP:SAMPLEPREP_SUMMARY            	equal aliquots and dried using CentriVap Concentrator System at 50 °C. One
SP:SAMPLEPREP_SUMMARY            	aliquot was used for phospholipids and the other for sphingolipids analysis.
SP:SAMPLEPREP_SUMMARY            	Phospholipids extraction For phospholipid extraction, the dried lipid film was
SP:SAMPLEPREP_SUMMARY            	desalted by butanol extraction using 300 µl of water-soluble butanol and 150
SP:SAMPLEPREP_SUMMARY            	µl of sterile distilled water. The mixture was vortexed and centrifuged. The
SP:SAMPLEPREP_SUMMARY            	top layer was pooled and dried in CentriVap Concentrator System at 4°C. The
SP:SAMPLEPREP_SUMMARY            	dried phospholipids were resuspended in 400 µl chloroform and methanol (1:1,
SP:SAMPLEPREP_SUMMARY            	v/v), vortexed for 30 s and were centrifuged again at 10,000 rpm for 5 min
SP:SAMPLEPREP_SUMMARY            	before injecting into liquid chromatography system. Sphingolipids extraction A
SP:SAMPLEPREP_SUMMARY            	fraction enriched in sphingolipids was obtained by mild alkaline hydrolysis,
SP:SAMPLEPREP_SUMMARY            	which degrades ester linkages found in many glycerophospholipids (Brockerhoff,
SP:SAMPLEPREP_SUMMARY            	1963). To achieve this, the dried lipid films were resuspended in 400 µl
SP:SAMPLEPREP_SUMMARY            	chloroform: methanol: water (16 : 16 : 5, v/v/v). Glycerophospholipids were
SP:SAMPLEPREP_SUMMARY            	deacylated by 400 µl of 0.2 N NaOH and incubated at 30 °C for 45 minutes. 400
SP:SAMPLEPREP_SUMMARY            	µl 0.5 M EDTA was added and the samples were neutralized with 80 µl of 1 N
SP:SAMPLEPREP_SUMMARY            	acetic acid. 400 µl of chloroform was added before the samples were vortexed
SP:SAMPLEPREP_SUMMARY            	and centrifuged. Sphingolipids were pooled by collecting the lower phase of the
SP:SAMPLEPREP_SUMMARY            	layers and it was dried using CentriVap Concentrator System at 4°C. The lipid
SP:SAMPLEPREP_SUMMARY            	extract was then desalted using butanol extraction as described above. Liquid
SP:SAMPLEPREP_SUMMARY            	Chromatography-Mass spectrometry (LC-MS) The LC-MS of the C. albicans lipids
SP:SAMPLEPREP_SUMMARY            	were performed using a 1260 Infinity High Performance Liquid Chromatography
SP:SAMPLEPREP_SUMMARY            	system coupled with a 6540 UHD Accurate-Mass Q-TOF mass spectrometer from
SP:SAMPLEPREP_SUMMARY            	Agilent Technologies with a Dual Agilent Jet Stream Electrospray Ionization
SP:SAMPLEPREP_SUMMARY            	(Dual AJS ESI) source. Typically, 2 µl of sample was injected for mass
SP:SAMPLEPREP_SUMMARY            	spectrometry analysis. The Dual AJS ESI capillary voltage and nozzle voltage was
SP:SAMPLEPREP_SUMMARY            	maintained at 3.0 kV and 1 kV, respectively. The gas temperature was maintained
SP:SAMPLEPREP_SUMMARY            	at 300 °C, drying gas flow was set at the rate of 8 L/min, sheath gas
SP:SAMPLEPREP_SUMMARY            	temperature and sheath gas flow at 350 °C and 11 L/min respectively and
SP:SAMPLEPREP_SUMMARY            	nebulizer pressure was set at 35 psi. The mass spectrum was acquired from a
SP:SAMPLEPREP_SUMMARY            	mass-to charge ratio (m/z) of 400–1400 in the positive and negative ion mode,
SP:SAMPLEPREP_SUMMARY            	with an acquisition time of 3 minutes, and the scan duration was 1 second.
SP:SAMPLEPREP_SUMMARY            	Samples were directly infused using an autosampler syringe pump at a flow rate
SP:SAMPLEPREP_SUMMARY            	of 10 µl/min into Zorbax Eclipse Plus C18, 2.1 x 100 mm, 1.8 µm reverse phase
SP:SAMPLEPREP_SUMMARY            	column. The mobile phase was chloroform and methanol with 1 : 1 (v/v ratio) and
SP:SAMPLEPREP_SUMMARY            	water with 0.1% formic acid at a flow rate of 15 µl/min. Individual molecular
SP:SAMPLEPREP_SUMMARY            	species was identified using tandem mass spectrometry and in general, the
SP:SAMPLEPREP_SUMMARY            	collision energy used was in the range 25–80 eV. Two reference masses were
SP:SAMPLEPREP_SUMMARY            	used in each ionization modes, i.e., 121.0509 m/z and 922.0098 m/z for positive
SP:SAMPLEPREP_SUMMARY            	ionization, 112.9855 m/z and 1033.9881 m/z for negative ionization mode. All the
SP:SAMPLEPREP_SUMMARY            	data attained from mass spectral was in a d. format.
SP:PROCESSING_STORAGE_CONDITIONS 	Described in summary
SP:EXTRACT_STORAGE               	Described in summary
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent 6530
CH:COLUMN_NAME                   	Agilent Zorbax Eclipse Plus C18 (100 x 2.1mm, 1.8 um)
CH:FLOW_GRADIENT                 	yes
CH:FLOW_RATE                     	15ul/ml
CH:COLUMN_TEMPERATURE            	45
CH:SOLVENT_A                     	Chloroform and methanol
CH:SOLVENT_B                     	Water and formic acids
CH:INTERNAL_STANDARD             	121.0509 m/z and 922.0098 m/z for positive ionization, 112.9855 m/z and
CH:INTERNAL_STANDARD             	1033.9881 m/z for negative ionization mode.
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Agilent 6540 QTOF
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Agilent MassHunter Workstation Qualitative Analysis software version B.06.00
MS:MS_RESULTS_FILE               	ST001982_AN003233_Results.txt	UNITS:minute	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END