VCU Lipidomics/Metabolomics Shared Resource
The VCU Lipidomics/Metabolomics Shared Resource (VLMC) is a key focal point for development of a sustainable critical mass of expertise in lipid signaling and state-of-the-art analytical techniques.
- Core Overview
The objective/goal of the VCU Lipidomics/Metabolomics Shared Resource (VLMC) is to provide cost-effective and highly quantitative lipid and small metabolite analyses for Massey Cancer Center members. The resource director and/or staff also provide: 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 a centralized resource located in newly renovated space on the 2nd floor of Sanger Hall in rooms 2-012 and 2-006 with additional space containing freezers for longer term storage in 2-010. The facility is within close walking distance of the Massey Cancer Center, and standard hours for the facility are 9:00 am until 5:00 pm, Monday through Friday, and certified users have access to the facility 24 hours per day, 7 days per week.
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 necessary 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 by the VLMC). The lipidomics facility provides both qualitative and quantitative analysis of lipid species utilizing 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 allowing us to analyze 157 of these lipids in a single run and for examining low abundant lipids and protein-lipid interactions due to their high sensitivity.
The VLMC has recently acquired an AB SCIEX Quadrupole Time of Flight (QTOF 5600+) mass analyzer. This high-resolution mass analyzer allows us to undertake investigative analysis of biological systems. A typical workflow would include 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. As such, 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 our existing complement of 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 opens the door for scientists at Massey to quickly and efficiently carry out research into diverse areas such as biomarker identification and validation from multiple biological material, identification of metabolic perturbation following various cancer treatments as well as comparative analysis of differential treatment regimes. We welcome the scientific community of VCU to utilize this resource for expanding their research into new, exciting and often unexplored territories in basic science as well as clinical and translational research. Indeed, this facility has led to >75 peer-reviewed publications including two Nature papers, a Science paper, and a paper in Cancer Cell. This facility has also aided VCU researchers in obtaining national awards as well as led to the funding of numerous extramural grant applications for VCU faculty. Overall, the VLMC is one of the most capable Lipidomics/Metabolomics facilities in the country.
Rates and member subsidy
Massey members pay 75 percent of these rates per the member subsidy program.
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
*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 our online system or via e-mail submission to Ms. Kubick with a Microsoft Word version of the VLMC Submission Form. 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