VCU Lipidomics/Metabolomics Shared Resource
The VCU Lipidomics/Metabolomics Core (VLMC) is a focal point for the development of a sustainable critical mass of expertise in lipid signaling and state-of-the-art analytical techniques.
- Core Overview
The objective of the VCU Lipidomics/Metabolomics Core (VLMC) is to provide cost-effective and quantitative lipid and small metabolite analyses for Massey Cancer Center members. Staff provides consultative services for experimental design and data interpretation, education to certify end-users in the use of the equipment, operator-assisted analysis, and instrument maintenance. The VLMC is located in Sanger Hall 2-012 and 2-006 with additional space and freezers in 2-010. Standard facility are 9:00 am - 5:00 pm Monday through Friday. Certified users have access to the facility 24/7.
The facility is built around four tandem triple quadrupole/linear ion trap mass spectrometers, two high-end AB SCIEX 4000 QTRAPs, an AB SCIEX 5500 QTRAP, and an AB SCIEX 6500 QTRAP. These instruments have the resolution and sensitivity for a broad range of qualitative and quantitative lipid and metabolite analyses required for exhaustive characterization of lipid signaling molecules and metabolic pathways (see included Table for lipid classes analyzed). The facility provides qualitative and quantitative analysis of lipid species using high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS). The AB SCIEX 4000 QTRAPs are mainly utilized for the analysis of high-level lipids (e.g. SM, S1P (in serum), ceramides, LPA, CL, etc.). The 5500 and 6500 QTRAPs are used for eicosanoids to analyze 157 lipids in a single run and to examine low abundant lipids and protein-lipid interactions due to their high sensitivity.
The VLMC has acquired an AB SCIEX Quadrupole Time of Flight (QTOF 5600+) mass analyzer, which allows users to conduct investigative analysis of biological systems. A typical workflow includes interrogation of the entire mass range of the analyzer (50-1250 m/z) at a mass accuracy of 1-2ppm between a treatment and control biological state via MS and MS/MS analysis. This data acquisition is followed by post-acquisition comparative statistical analysis to identify the changes in the metabolome/lipidome between the two states. This mass analyzer is an effective hypotheses generator, allowing the identification of perturbations in the metabolome/lipidome during the onset and progression of disease as well as with different treatments. In conjunction with the high throughput hybrid triple quadruple linear ion trap mass analyzers, the generated hypotheses can easily be validated in large sample cohorts in a targeted fashion.
The integrated workflow of the VLMC allows scientists to efficiently carry out research in diverse areas, such as biomarker identification and validation from multiple biological material, identification of metabolic perturbation following various cancer treatments, and comparative analysis of differential treatment regimes.
Current rates (per sample and includes sample processing):
VCU Faculty Cost
Massey Cancer Center Member Cost
Non-profit, Non-VCU Cost
TA: Steroid Hormones
TA: Lysophosphatidic Acid
TA: Vitamin D
UTA: Systems Level Lipidomics
UTA: Systems Level Metabolomics
TA: targeted, quantitative analysis; UTA: untargeted, qualitative analysis
Massey member subsidy benefit
Massey subsidizes 25 percent of the cost of members’ core laboratory usage for the following Massey-supported, fee-for-service shared resources:
- Flow Cytometry Shared Resource
- VCU Lipidomics/Metabolomics Shared Resource
- Structural Biology Shared Resource
- Tissue and Data Acquisition and Analysis Core
- Transgenic/Knock-out Mouse Facility Shared Resource
- Microscopy Shared Resource
To receive this member benefit, please initiate core lab services using a VCU index code associated with your name. When you are billed for core laboratory usage, the Rams FORCE system will automatically split the invoice between your index code and a Massey account established for this purpose. You will be charged 75 percent of the published rate and Massey will provide a 25 percent co-pay to cover the balance. Please direct any questions or concerns about the member subsidy benefit to Paul Fawcett, Ph.D. at firstname.lastname@example.org or (804) 833-1959.
*Reduced rates and pricing are also available for VLMC certified users at a No-technician rate.
Before submitting samples, you must contact VLMC personnel to set up appropriate logistics. Some lipids are unstable and require immediate analysis. Our goal is to provide you with the best data possible! All data will be delivered to the submitting individual and laboratory PI as an easy-to-read Excel file. We are also happy to liaise with other Massey Cancer Center core facilities, such as the Biostatistics Core. Please note, all samples must be accompanied by a sample submission form via e-mail to Ms. Kubick with a Microsoft Word version of the VLMC Submission Form internal Feb 2020 or VLMC Submission Form external Feb 2020. Please note that normalization is required. Dr. Jeremy Allegood should be contacted for all analyses.
Eicosanoid analysis: From serum, plasma, tissues, cells, cell media (dye-free preferred), lavages, etc…, we concentrate via a SPE column followed by the quantitative, targeted analysis of 30 analytes using deuterated standards.
These include: 6-keto Prostaglandin F1α, 8-iso Prostaglandin F2α, Thromboxane B2, 5-iPF2α-VI, Prostaglandin E2, Prostaglandin F2α, Prostaglandin D2, Resolvin D2, Prostaglandin E1, Resolvin D1, Lipoxin A4, Prostaglandin A2, Leukotriene B4, Leukotriene C4, Leukotriene E4, Leukotriene D4, (±)14,15-DHET, 15-deoxy-Δ12,14-Prostaglandin J2, (±)11,12-DHET, (±)8,9-DHET, 20-HETE, 15 HETE, 12 HETE, (±)14(15)-EET, 5 HETE, (±)8(9)-EET, Eicosapentaenoic Acid, Docosahexaenoic Acid, Arachidonic Acid, and Dihomo-Gamma-Linolenic Acid
Coming soon: We will also be offering a semi-quantitative, “relative change” analysis, which will include: Resolvin E1, maresin 1, Prostaglandin D3, Prostaglandin E3, Resolvin D5, Resolvin D3, maresin, 2,10(S),17(S)-DiHDoHE (PDX), Prostaglandin J2. As 13C and deuterated standards are not available for these analytes, we use the closely related PGE2, PGE3, and Resolvin D2 internal standards (deuterated). These data are considered quantitative for identifying fold-changes between provided samples, but the actual quantitation pmols/mL, etc… are considered semi-quantitative. As deuterated or 13C standards become available for these compounds, they will be added to our full quantitative analysis package for no additional charge.
Sphingolipid analysis: From serum, plasma, tissues, cells, cell media (dye-free preferred), lavages, etc…, we extract samples using a modified Bligh-Dyer followed by the quantitative, targeted analysis of:
Ceramide (N-acyl chain lengths = C14:0, C16:0, C18:1, C18:0, C20:0, C22:0, C24:1, C24:0, C26:1, C26:0)
Monohexosylceramide (N-acyl chain lengths = C14:0, C16:0, C18:1, C18:0, C20:0, C22:0, C24:1, C24:0, C26:1, C26:0)
Sphingomyelin (N-acyl chain lengths = C14:0, C16:0, C18:1, C18:0, C20:0, C22:0, C24:1, C24:0, C26:1, C26:0)
Ceramide 1-Phosphate (N-acyl chain lengths = C14:0, C16:0, C18:1, C18:0, C20:0, C22:0, C24:1, C24:0, C26:1, C26:0) (Of note: C1Ps are labile and usually require “fresh” samples for appropriate detection and quantitation).
Coming soon: As soon as scientific/biological validation is complete, we will be offering deoxysphingolipids as well.
Steroid analysis: We extract samples using a modified Bligh-Dyer followed by the quantitative, targeted analysis of:
Dehydroepiandrosterone, Androstenedione, Testosterone, Estradiol, 2-Methoxyestradiol, Dihydrotestosterone, Cortisol, Progesterone, and Corticosterone.
Cardiolipin analysis: We extract samples using a modified Bligh Dyer followed by the quantitative analysis of the following: (C14:0)4, (C18:2)3(C18:3)1, (C18:2)4, (C18:2)3(C18:1)1, (C18:2)2(C18:1)2, (C18:2)3(C18:0)1, (C18:2)3(C22:6)1, (C18:2)2(C18:1)1(C22:6)1, and (C18:2)2(C22:6)2.
Coming soon: As soon as scientific/biological validation is complete, we will be offering certain oxidized cardiolipin species as well.
Lysophosphatidic acid analysis: We extract samples using a modified Bligh-Dyer followed by the quantitative, targeted analysis of: (16:0, 18:2, 18:1, 18:0, and 20:4).
Systems level, untargeted lipid analysis: We extract samples using a modified Bligh-Dyer followed by the quantitative analysis of multiple lipid classes e.g; (Glycerophospholipids, Sphingolipids, Glycerolipids, Sterols, & Fatty Acids). Plasma analysis provides approximately 2000 positive and 2000 negative analytes.
Phospholipid Analysis: We extract samples using a modified Bligh-Dyer followed by the quantitative, targeted analysis of:
PE’s (16:0-20:4 PE, 16:0-22:6 PE, 18:0-20:4 PE, 18:0-22:6 PE, 16:0 Lyso PE, 18:0 Lyso PE, EPA, DHA, & AA)
PC’s (16:0-20:4 PC, 16:0-22:6 PC, 16:0 Lyso PC, 18:0-20:4 PC, 18:0-22:6 PC, & 18:0 Lyso PC)
Urine Metabolites: We concentrate via a SPE column followed by the quantitative, targeted analysis of: (2,3-dinor-6-keto PGF1alpha-d9, 2,3-dinor TXB2-d9, 11-dehydro TXB2-d4, 2,3-dinor-6-keto PGF1alpha, 11-dehydro-2,3-dinor TXB2, 2,3-dinor TXB2, & 11-dehydro TXB2).
These will be added soon to the Eicosanoid Analysis at no additional charge.
Vitamin D Analysis: We extract samples using a LLE method followed by the quantitative, targeted analysis of: (Vitamin D3, 25-hydroxyvitamin D3, 3-epi-25-hydroxyvitamin D3, & 24R,25-dihydroxyvitamin D3).
Cholesterol & Cholesterol Esters: In process
VLMC scientific and executive director
L. Ashley Cowart, Ph.D
Department of Biochemistry and Molecular Biology
VLMC operational director/manager
Jeremy Allegood, Ph.D.
Department of Biochemistry and Molecular Biology
Sanger Hall; Rooms 2-006 and 2-012
VLMC laboratory specialist
Sanger Hall; Rooms 2-006 and 2-012
Suggested grant language
The VCU Lipidomics and Metabolomics Core Laboratory (updated June 2019) The Virginia Commonwealth University (VCU) Lipidomics & Metabolomics Core Laboratory (VLMC) provides highly sensitive and specific analyses of lipids and small-metabolites for investigators at VCU and Massey Cancer Center. The VLMC facilities enable the qualitative and quantitative investigation of the lipidome in both in vitro and in vivo studies involving cell lines, animal models, and/or human samples. This is accomplished using an advanced lipidomic instrumentation suite that is nationally competitive. Specifically, the VLMC maintains 3 advanced quadrupole linear iontrap (QTrap) mass spectrometers, each equipped with an ultra-high performance liquid chromatography (UPLC) system. The core also utilizes a Thermoscientific Q Exactive HF OrbiTrap coupled to a UPLC for untargeted lipidomics and metabolomics with 240,000 resolution. This equipment enables highly sensitive and specific identification and/or quantification of changes to lipids in a high-throughput fashion and over a large dynamic range. Lipid classes analyzed include glycerophospholipids, sphingolipids, sterols, and others; custom analyses are available upon consultation. Untargeted analyses provide data from a database of over 8000 lipids and more than 20000 water-soluble metabolites. The resource is directed by L. Ashley Cowart, PhD, who is supported by the operational director, Jeremy Allegood, PhD, and an additional FTE. Drs. Allegood and Cowart provide consultative services for experimental design and data interpretation and assist with novel method development. The VLMC is a jointly managed (MCC and VCU) resource located in Sanger Hall and within close walking distance of MCC. Standard hours for the facility are 9:00 AM until 5:00 PM, Monday through Friday. The usage of services and equipment is tracked and charged back. The VLMC adds significant value to VCU and MCC research by providing services that otherwise would not be practically obtainable or would be cost prohibitive through alternative vendors or even other non-profit institutions. Overall, the instrumentation and expertise available in the VLMC have facilitated the development of a growing user base of VCU faculty and MCC members, and investigators from other institutions whose research routinely garners peer-reviewed national funding and is published in high-impact scientific journals.
Major instrumentation in the core includes:
Sciex 4000 QTrap: Quadrupole Linear Ion Trap used primarily for high abundance lipids from tissue or cell samples (1 million or more cells). Utilized for sphingolipids and glycerophospholipids. LoQ for most lipids between 1-10 pmol. Sciex 5500 QTrap: Quadrupole Linear Ion Trap used for most of VLMC’s quantitative lipid methods. Allows for 100 fmol or lower LoQ for sphingolipids, eicosanoids, glycerophospholipids, steroids, and lysophospholipids. Allows of analysis of samples that range from lipid bound to protein pulldowns to cells/tissues/fluids.
Sciex 6500 QTrap: Quadrupole Linear Ion Trap used for VLMC’s quantitative lipid methods that require more sensitivity in negative mode or of higher m/z values. Beneficial for cardiolipins, sulfatides, ganglioside species, and some glycerophospholipids. Thermoscientific Q Exactive HF OrbiTrap: This is the VLMC’s newest instrument for lipidomics and metabolomics. The OrbiTrap allows identification and relative quantitation of over 8000 lipid species and more than 20000 water-soluble metabolites. Utilizing the OrbiTrap’s 240000 resolution allows significantly increased confidence in qualitative data.