Summary of study ST001982

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001258. The data can be accessed directly via it's Project DOI: 10.21228/M83M69 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001982
Study TitleLipidomic characterization of Candida albicans in response to Aureobasidin treatment in vitro.
Study SummaryCandida albicans is an opportunistic yeast pathogen that causes a wide range of infections especially amongst immunocompromised patients. Aureobasidin A (AbA) has been shown to inhibit inositolphosphoryl ceramide synthase (IPCS), a key enzyme responsible for sphingolipid biosynthesis. There are limited studies exploring IPCS as a target molecule for antifungal treatment. It is hypothesized that the mechanism of AbA inhibition involves alteration of C. albicans phospholipid and sphingolipid profiles. The profiling of C. albicans phospholipid and sphingolipid upon exposure to 0.5-4 µg/ml of AbA were determined using Liquid chromatography-mass spectrometry (LC-MS).
Institute
University of Malaya
Last NameHamdan
First NameNur Wahida
AddressJalan Profesor Diraja Ungku Aziz, 50603 Kuala Lumpur, Wilayah Persekutuan Kuala Lumpur, Malaysia
Emailnurwahidahamdan@siswa.um.edu.my
Phone0193354272
Submit Date2021-09-16
Num Groups5
Total SubjectsDuplicates
Num MalesNA
Num FemalesNA
Analysis Type DetailLC-MS
Release Date2021-11-22
Release Version1
Nur Wahida Hamdan Nur Wahida Hamdan
https://dx.doi.org/10.21228/M83M69
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001258
Project DOI:doi: 10.21228/M83M69
Project Title:Lipidomic characterization of Candida albicans in response to Aureobasidin treatment in vitro.
Project Summary:Candida albicans is an opportunistic yeast pathogen that causes a wide range of infections especially amongst immunocompromised patients. Aureobasidin A (AbA) has been shown to inhibit inositolphosphoryl ceramide synthase (IPCS), a key enzyme responsible for sphingolipid biosynthesis. There are limited studies exploring IPCS as a target molecule for antifungal treatment. It is hypothesized that the mechanism of AbA inhibition involves alteration of C. albicans phospholipid and sphingolipid profiles. The profiling of C. albicans phospholipid and sphingolipid upon exposure to 0.5-4 µg/ml of AbA were determined using Liquid chromatography-mass spectrometry (LC-MS).
Institute:University of Malaya
Department:Medical Microbiology Department
Laboratory:Lab 3
Last Name:Hamdan
First Name:Nur Wahida
Address:Jalan Profesor Diraja Ungku Aziz, 50603 Kuala Lumpur, Wilayah Persekutuan Kuala Lumpur, Malaysia
Email:nurwahidahamdan@siswa.um.edu.my
Phone:0193354272
Funding Source:FRGS(FP035-2014A), UM PPP(PG178-2015B)

Subject:

Subject ID:SU002063
Subject Type:Yeast
Subject Species:Candida albicans
Taxonomy ID:5476
Genotype Strain:SC 5314

Factors:

Subject type: Yeast; Subject species: Candida albicans (Factor headings shown in green)

mb_sample_id local_sample_id Treatment Concentration (ug/ml) Extraction method Ionization mode
SA185589n0.5_PBAbA-treated 0.5 Phospholipids Negative
SA185590n0.5_PAAbA-treated 0.5 Phospholipids Negative
SA185591N_0.5_PAAbA-treated 0.5 Phospholipids Negative
SA185592N_0.5_PBAbA-treated 0.5 Phospholipids Negative
SA185593p0.5_PBAbA-treated 0.5 Phospholipids Positive
SA185594p0.5_PAAbA-treated 0.5 Phospholipids Positive
SA185595P_0.5_PBAbA-treated 0.5 Phospholipids Positive
SA185596P_0.5_PAAbA-treated 0.5 Phospholipids Positive
SA185597N_0.5_SBAbA-treated 0.5 Sphingolipids Negative
SA185598n0.5_SAAbA-treated 0.5 Sphingolipids Negative
SA185599n0.5_SBAbA-treated 0.5 Sphingolipids Negative
SA185600N_0.5_SAAbA-treated 0.5 Sphingolipids Negative
SA185601p0.5_SBAbA-treated 0.5 Sphingolipids Positive
SA185602p0.5_SAAbA-treated 0.5 Sphingolipids Positive
SA185603P_0.5_SBAbA-treated 0.5 Sphingolipids Positive
SA185604P_0.5_SAAbA-treated 0.5 Sphingolipids Positive
SA185617N_1_PAAbA-treated 1 phospholipids Negative
SA185618n1_PAAbA-treated 1 phospholipids Negative
SA185605n1_PBAbA-treated 1 Phospholipids Negative
SA185606N_1_PBAbA-treated 1 Phospholipids Negative
SA185619p1_PAAbA-treated 1 phospholipids Positive
SA185620P_1_PAAbA-treated 1 phospholipids Positive
SA185607P_1_PBAbA-treated 1 Phospholipids Positive
SA185608p1_PBAbA-treated 1 Phospholipids Positive
SA185609n1_SAAbA-treated 1 Sphingolipids Negative
SA185610n1_SBAbA-treated 1 Sphingolipids Negative
SA185611N_1_SAAbA-treated 1 Sphingolipids Negative
SA185612N_1_SBAbA-treated 1 Sphingolipids Negative
SA185613P_1_SAAbA-treated 1 Sphingolipids Positive
SA185614p1_SBAbA-treated 1 Sphingolipids Positive
SA185615P_1_SBAbA-treated 1 Sphingolipids Positive
SA185616p1_SAAbA-treated 1 Sphingolipids Positive
SA185621N_2_PAAbA-treated 2 Phospholipids Negative
SA185622n2_PBAbA-treated 2 Phospholipids Negative
SA185623n2_PAAbA-treated 2 Phospholipids Negative
SA185624N_2_PBAbA-treated 2 Phospholipids Negative
SA185625P_2_PBAbA-treated 2 Phospholipids Positive
SA185626P_2_PAAbA-treated 2 Phospholipids Positive
SA185627p2_PBAbA-treated 2 Phospholipids Positive
SA185628p2_PAAbA-treated 2 Phospholipids Positive
SA185629n2_SAAbA-treated 2 Sphingolipids Negative
SA185630N_2_SAAbA-treated 2 Sphingolipids Negative
SA185631N_2_SBAbA-treated 2 Sphingolipids Negative
SA185632n2_SBAbA-treated 2 Sphingolipids Negative
SA185633P_2_SAAbA-treated 2 Sphingolipids Positive
SA185634p2_SAAbA-treated 2 Sphingolipids Positive
SA185635p2_SBAbA-treated 2 Sphingolipids Positive
SA185636P_2_SBAbA-treated 2 Sphingolipids Positive
SA185637N_4_PBAbA-treated 4 Phospholipids Negative
SA185638n4_PBAbA-treated 4 Phospholipids Negative
SA185639N_4_PAAbA-treated 4 Phospholipids Negative
SA185640n4_PAAbA-treated 4 Phospholipids Negative
SA185641p4_PAAbA-treated 4 Phospholipids Positive
SA185642p4_PBAbA-treated 4 Phospholipids Positive
SA185643P_4_PAAbA-treated 4 Phospholipids Positive
SA185644P_4_PBAbA-treated 4 Phospholipids Positive
SA185645N_4_SAAbA-treated 4 Sphingolipids Negative
SA185646N_4_SBAbA-treated 4 Sphingolipids Negative
SA185647n4_SBAbA-treated 4 Sphingolipids Negative
SA185648n4_SAAbA-treated 4 Sphingolipids Negative
SA185649P_4_SAAbA-treated 4 Sphingolipids Positive
SA185650p4_SAAbA-treated 4 Sphingolipids Positive
SA185651p4_SBAbA-treated 4 Sphingolipids Positive
SA185652P_4_SBAbA-treated 4 Sphingolipids Positive
SA185653nDMSO_PAControl - Phospholipids Negative
SA185654nDMSO_PBControl - Phospholipids Negative
SA185655N_DMSO_PAControl - Phospholipids Negative
SA185656N_DMSO_PBControl - Phospholipids Negative
SA185657pDMSO_PAControl - Phospholipids Positive
SA185658pDMSO_PBControl - Phospholipids Positive
SA185659P_DMSO_PAControl - Phospholipids Positive
SA185660P_DMSO_PBControl - Phospholipids Positive
SA185661nDMSO_SBControl - Sphingolipids Negative
SA185662nDMSO_SAControl - Sphingolipids Negative
SA185663N_DMSO_SAControl - Sphingolipids Negative
SA185664N_DMSO_SBControl - Sphingolipids Negative
SA185665P_DMSO_SBControl - Sphingolipids Positive
SA185666pDMSO_SBControl - Sphingolipids Positive
SA185667pDMSO_SAControl - Sphingolipids Positive
SA185668P_DMSO_SAControl - Sphingolipids Positive
Showing results 1 to 80 of 80

Collection:

Collection ID:CO002056
Collection Summary:A starter culture was prepared by inoculating two loopfuls of C. albicans yeast colony in 5 ml of yeast peptone dextrose (YPD) broth and incubated at 37°C for 24 hours. 150 microlitres of the starter culture was then inoculated into 150 ml fresh YPD broth (10^7 of cells/ml) and let to grow for 6 hours until it reached 10^9 of cells/ml.
Sample Type:Yeast cells
Storage Conditions:-80℃

Treatment:

Treatment ID:TR002075
Treatment Summary:The yeast cells were exposed to different concentration of Aureobasidin A (0.5, 1, 2 and 4 microgram/ml). DMSO-treated yeast culture was used as a control. The cultures were incubated for another 3 hours prior to harvesting. Each of the conditions was performed in duplicate.

Sample Preparation:

Sampleprep ID:SP002069
Sampleprep Summary:Lipids enrichment Lipids were enriched using a method as described by Guan and Wenk (2010). After AbA treatment, the yeast cells were harvested and washed twice. The wet weight was standardized. Briefly, the cells were resuspended in 2 ml 95% ethanol: water: diethyl ether: pyridine: ammonium hydroxide (15 : 15 : 5 : 1 : 0.018). The cells were broken by glass beads (vortexed twice for 1 minute each) and incubated for 20 min at 60 °C. Debris was pelleted by centrifugation and the supernatant was transferred to a fresh tube. The pellet was re-extracted once more using the same procedure. The pooled supernatants were divided into equal aliquots and dried using CentriVap Concentrator System at 50 °C. One aliquot was used for phospholipids and the other for sphingolipids analysis. Phospholipids extraction For phospholipid extraction, the dried lipid film was desalted by butanol extraction using 300 µl of water-soluble butanol and 150 µl of sterile distilled water. The mixture was vortexed and centrifuged. The top layer was pooled and dried in CentriVap Concentrator System at 4°C. The dried phospholipids were resuspended in 400 µl chloroform and methanol (1:1, v/v), vortexed for 30 s and were centrifuged again at 10,000 rpm for 5 min before injecting into liquid chromatography system. Sphingolipids extraction A fraction enriched in sphingolipids was obtained by mild alkaline hydrolysis, which degrades ester linkages found in many glycerophospholipids (Brockerhoff, 1963). To achieve this, the dried lipid films were resuspended in 400 µl chloroform: methanol: water (16 : 16 : 5, v/v/v). Glycerophospholipids were deacylated by 400 µl of 0.2 N NaOH and incubated at 30 °C for 45 minutes. 400 µl 0.5 M EDTA was added and the samples were neutralized with 80 µl of 1 N acetic acid. 400 µl of chloroform was added before the samples were vortexed and centrifuged. Sphingolipids were pooled by collecting the lower phase of the layers and it was dried using CentriVap Concentrator System at 4°C. The lipid extract was then desalted using butanol extraction as described above. Liquid Chromatography-Mass spectrometry (LC-MS) The LC-MS of the C. albicans lipids were performed using a 1260 Infinity High Performance Liquid Chromatography system coupled with a 6540 UHD Accurate-Mass Q-TOF mass spectrometer from Agilent Technologies with a Dual Agilent Jet Stream Electrospray Ionization (Dual AJS ESI) source. Typically, 2 µl of sample was injected for mass spectrometry analysis. The Dual AJS ESI capillary voltage and nozzle voltage was maintained at 3.0 kV and 1 kV, respectively. The gas temperature was maintained at 300 °C, drying gas flow was set at the rate of 8 L/min, sheath gas temperature and sheath gas flow at 350 °C and 11 L/min respectively and nebulizer pressure was set at 35 psi. The mass spectrum was acquired from a mass-to charge ratio (m/z) of 400–1400 in the positive and negative ion mode, with an acquisition time of 3 minutes, and the scan duration was 1 second. Samples were directly infused using an autosampler syringe pump at a flow rate of 10 µl/min into Zorbax Eclipse Plus C18, 2.1 x 100 mm, 1.8 µm reverse phase column. The mobile phase was chloroform and methanol with 1 : 1 (v/v ratio) and water with 0.1% formic acid at a flow rate of 15 µl/min. Individual molecular species was identified using tandem mass spectrometry and in general, the collision energy used was in the range 25–80 eV. Two reference masses were used in each ionization modes, i.e., 121.0509 m/z and 922.0098 m/z for positive ionization, 112.9855 m/z and 1033.9881 m/z for negative ionization mode. All the data attained from mass spectral was in a d. format.
Processing Storage Conditions:Described in summary
Extract Storage:Described in summary

Combined analysis:

Analysis ID AN003232 AN003233
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Agilent 6530 Agilent 6530
Column Agilent Zorbax Eclipse Plus C18 (100 x 2.1mm, 1.8 um) Agilent Zorbax Eclipse Plus C18 (100 x 2.1mm, 1.8 um)
MS Type ESI ESI
MS instrument type QTOF QTOF
MS instrument name Agilent 6540 QTOF Agilent 6540 QTOF
Ion Mode POSITIVE NEGATIVE
Units minute minute

Chromatography:

Chromatography ID:CH002384
Instrument Name:Agilent 6530
Column Name:Agilent Zorbax Eclipse Plus C18 (100 x 2.1mm, 1.8 um)
Column Temperature:45
Flow Gradient:yes
Flow Rate:15ul/ml
Internal Standard:121.0509 m/z and 922.0098 m/z for positive ionization, 112.9855 m/z and 1033.9881 m/z for negative ionization mode.
Solvent A:Chloroform and methanol
Solvent B:Water and formic acids
Chromatography Type:Reversed phase

MS:

MS ID:MS003006
Analysis ID:AN003232
Instrument Name:Agilent 6540 QTOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:Agilent MassHunter Workstation Qualitative Analysis software version B.06.00
Ion Mode:POSITIVE
  
MS ID:MS003007
Analysis ID:AN003233
Instrument Name:Agilent 6540 QTOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:Agilent MassHunter Workstation Qualitative Analysis software version B.06.00
Ion Mode:NEGATIVE
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