Summary of Study ST003195

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 PR001991. The data can be accessed directly via it's Project DOI: 10.21228/M8CB2H This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study ID	ST003195
Study Title	Unveiling cellular changes in leukaemia cell line K-562 after cannabidiol treatment through lipidomics
Study Summary	The present study was aimed at revealing the metabolic changes that occurred in the cellular lipid pattern of acute and chronic myeloid leukaemia cells following treatment with cannabidiol (CBD). CBD is a non-psychoactive compound present in Cannabis sativa L., which has shown an antiproliferative action in these type of cancer cells, to determine significant alterations of the cell metabolism attributable to the induction of apoptosis, previously observed from in vitro studies. Control and treated cells of chronic myeloid leukaemia (K562) were studied through an untargeted lipidomics approach. Treatment was carried out with CBD at a concentration of 23 µM CBD for 48 h. After the extraction of the lipid content from cell lysates, the samples were analysed by UHPLC-QTOF-MS/MS both in the positive and the negative ionization modes.
Institute	Universidad CEU San Pablo
Laboratory	CEMBIO
Last Name	Garcia Fernández
First Name	Antonia
Address	Urb. Montepríncipe., Alcorcón, Madrid, 28925, Spain
Email	antogar@ceu.es
Phone	(+34) 91 372 47 69
Submit Date	2024-05-08
Num Groups	2
Total Subjects	10
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	LC-MS
Release Date	2024-09-17
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR001991
Project DOI:	doi: 10.21228/M8CB2H
Project Title:	Unveiling cellular changes in leukaemia cell line K-562 after cannabidiol treatment through lipidomics
Project Summary:	The present study was aimed at revealing the metabolic changes that occurred in the cellular lipid pattern of acute and chronic myeloid leukaemia cells following treatment with cannabidiol (CBD). CBD is a non-psychoactive compound present in Cannabis sativa L., which has shown an antiproliferative action in these type of cancer cells, to determine significant alterations of the cell metabolism attributable to the induction of apoptosis, previously observed from in vitro studies. Control and treated cells of chronic myeloid leukaemia (K562) were studied through an untargeted lipidomics approach. Treatment was carried out with CBD at a concentration of 23 µM CBD for 48 h. After the extraction of the lipid content from cell lysates, the samples were analysed by UHPLC-QTOF-MS/MS both in the positive and the negative ionization modes.
Institute:	Universidad CEU San Pablo
Laboratory:	CEMBIO
Last Name:	Garcia Fernández
First Name:	Antonia
Address:	Urb. Montepríncipe., Alcorcón, Madrid, 28925, Spain
Email:	antogar@ceu.es
Phone:	(+34) 91 372 47 69

Subject:

Subject ID:	SU003314
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Gender:	Not applicable

Factors:

Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id	local_sample_id	Sample source	Treatment
SA347951	K562_CBD1	Leukemia cell line K-562	CBD
SA347952	K562_CBD5	Leukemia cell line K-562	CBD
SA347953	K562_CBD4	Leukemia cell line K-562	CBD
SA347954	K562_CBD2	Leukemia cell line K-562	CBD
SA347955	K562_CBD3	Leukemia cell line K-562	CBD
SA347956	K562_CTR5	Leukemia cell line K-562	CTR
SA347957	K562_CTR4	Leukemia cell line K-562	CTR
SA347958	K562_CTR1	Leukemia cell line K-562	CTR
SA347959	K562_CTR2	Leukemia cell line K-562	CTR
SA347960	K562_CTR3	Leukemia cell line K-562	CTR

Showing results 1 to 10 of 10

Showing results 1 to 10 of 10

Collection:

Collection ID:	CO003307
Collection Summary:	K-562 cells were grown in RPMI medium supplemented with 10% FBS, 2 mM L-glutamine, 100 U/mL penicillin and 100 μg/mL streptomycin. Cells were cultured in a humidiﬁed incubator at 37 °C with 95% humidity and 5% CO2.Cells were treated with CBD. Cells were washed with cold PBS, centrifuged and the pellets were stored at - 80°C until the day of the analysis.
Sample Type:	Leukemia cells

Treatment:

Treatment ID:	TR003323
Treatment Summary:	Cells were treated with 23 µM of CBD

Sample Preparation:

Sampleprep ID:	SP003321
Sampleprep Summary:	The samples were vortex-mixed for 2 min and 100 µL of cold MeOH (- 20°C) were added to each replicate for deproteinization. Ultrasound probe (UP 200 S Dr. Hielscher Ultrasonic Gmbh) was used (0.5 cycles; 80 amplitude x 16 times) to lyse the cells. Cell lysis was verified using a microscope. Twenty µL of the working solution of 20 mg/L of C17 sphinganine in methanol were added to 100 µL of each replicate. After that, 284 µL of methyl tert-butyl ether (MTBE) was added to each sample to extract hydrophobic compounds. Each replicate were vortex-mixed for 1 h (room temperature, 10,000 xg). Then, 71 µL of water was added to each sample and vortex-mixed for 1 min (25°C, 10,000 xg). Samples were centrifuged for 10 min at 16,000 xg at 4 °C. After centrifugation, 90 µL of the supernatant were transferred to vials for the analysis by LC-MS in the positive and in the negative ionization mode

Sampleprep ID:

SP003321

Sampleprep Summary:

The samples were vortex-mixed for 2 min and 100 µL of cold MeOH (- 20°C) were added to each replicate for deproteinization. Ultrasound probe (UP 200 S Dr. Hielscher Ultrasonic Gmbh) was used (0.5 cycles; 80 amplitude x 16 times) to lyse the cells. Cell lysis was verified using a microscope. Twenty µL of the working solution of 20 mg/L of C17 sphinganine in methanol were added to 100 µL of each replicate. After that, 284 µL of methyl tert-butyl ether (MTBE) was added to each sample to extract hydrophobic compounds. Each replicate were vortex-mixed for 1 h (room temperature, 10,000 xg). Then, 71 µL of water was added to each sample and vortex-mixed for 1 min (25°C, 10,000 xg). Samples were centrifuged for 10 min at 16,000 xg at 4 °C. After centrifugation, 90 µL of the supernatant were transferred to vials for the analysis by LC-MS in the positive and in the negative ionization mode

Combined analysis:

Analysis ID	AN005243	AN005244
Analysis type	MS	MS
Chromatography type	Reversed phase	Reversed phase
Chromatography system	Agilent 1290 Infinity II	Agilent 1290 Infinity II
Column	Agilent InfinityLab Poroshell 120 EC-C18 (100 x 3mm,2.7um)	Agilent InfinityLab Poroshell 120 EC-C18 (100 x 3mm,2.7um)
MS Type	ESI	ESI
MS instrument type	QTOF	QTOF
MS instrument name	Agilent 6545 QTOF	Agilent 6545 QTOF
Ion Mode	POSITIVE	NEGATIVE
Units	Corrected areas	Corrected areas

Chromatography:

Chromatography ID:	CH003970
Instrument Name:	Agilent 1290 Infinity II
Column Name:	Agilent InfinityLab Poroshell 120 EC-C18 (100 x 3mm,2.7um)
Column Temperature:	50 °C
Flow Gradient:	Started at 70% of B at 0-1 min, 86% B at 3.5-10 min, 100% B at 11-17 min. The starting conditions were recovered by minute 17.
Flow Rate:	0.6 mL/min
Internal Standard:	Sphinganine (D17:0)
Solvent A:	90% Water, 10% Methanol; 10 mM ammonium acetate, 0.2 mM ammonium fluoride
Solvent B:	50% Isopropanol, 30% Methanol, 20% Acetonitrile; 10 mM ammonium acetate, 0.2 mM ammonium fluoride
Chromatography Type:	Reversed phase

MS:

MS ID:	MS004976
Analysis ID:	AN005243
Instrument Name:	Agilent 6545 QTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	The Agilent 6545 QTOF mass spectrometer equipped with a dual AJS ESI ion source was set with the following parameters: 3500 V in the positive ionization mode, 150 V fragment voltage, 50 psi nebulizer gas pressure, 12 L/min at 300°C drying gas flow rate in positive ionization mode, 750 V octopole radio frequency voltage and 65 V skimmer. Data were collected in positive ESI mode, operated in full scan mode from 40 to 1700 m/z with a scan rate of 3 spectra/s. The reference solution contained purine (C5H4N4) at m/z 121.0509 and HP-0921 (C18H18O6N3P3F24) at m/z 922.0098. These masses were continuously infused into the system through an Agilent 1260 Iso Pump at a 1 mL/min (split ratio 1:100) to provide a constant mass correction. Data was acquired using Agilent MassHunter Workstation Software LC/MS Data Acquisition for 6200 series TOF/6500 series Q-TOF B 9.0.9044.0 (Agilent Technologies). The raw data were processed using Agilent Technologies MassHunter Profinder B.10.0.2.162 (Santa Clara, United States) to clean the background noise and unrelated ions.
Ion Mode:	POSITIVE

MS ID:	MS004977
Analysis ID:	AN005244
Instrument Name:	Agilent 6545 QTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	The Agilent 6545 QTOF mass spectrometer equipped with a dual AJS ESI ion source was set with the following parameters: 3000 V in the positive ionization mode, 150 V fragment voltage, 50 psi nebulizer gas pressure, 10 L/min at 300°C drying gas flow rate in negative ionization mode, 750 V octopole radio frequency voltage and 65 V skimmer. Data were collected in positive ESI mode, operated in full scan mode from 40 to 1700 m/z with a scan rate of 3 spectra/s. The reference solution contained purine (C5H4N4) at m/z 119.0363 and HP-0921+ acetate (C18H18O6N3P3F24) at m/z 980.0163. These masses were continuously infused into the system through an Agilent 1260 Iso Pump at a 1 mL/min (split ratio 1:100) to provide a constant mass correction. Data was acquired using Agilent MassHunter Workstation Software LC/MS Data Acquisition for 6200 series TOF/6500 series Q-TOF B 9.0.9044.0 (Agilent Technologies). The raw data were processed using Agilent Technologies MassHunter Profinder B.10.0.2.162 (Santa Clara, United States) to clean the background noise and unrelated ions.
Ion Mode:	NEGATIVE