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Mevalonate Pathway and Isoprenoid Analysis Service

The mevalonate (MVA) pathway is a vital metabolic route that fuels the biosynthesis of sterols, ubiquinones, dolichols, and other isoprenoid lipids in mammals, plants, and microorganisms. At Creative Proteomics, we provide targeted LC–MS/MS and optional FTMS-based analysis to precisely quantify MVA and MEP intermediates across diverse matrices. Our service helps researchers pinpoint metabolic bottlenecks, evaluate enzyme regulation, and link lipid biosynthesis with mitochondrial and chloroplast functions for system-level biological interpretation.

We Help You Solve:

  • Identify control points in isoprenoid and sterol metabolism
  • Quantify mevalonate and MEP intermediates across species and matrices
  • Reveal functional links between redox lipids, prenylation, and membrane biology

Our Analytical Advantages:

  • LC–MS/MS linearity R2 ≥ 0.995 and CV ≤ 15% for qualified metabolites
  • Optional Orbitrap FTMS confirmation with ≤ 5 ppm mass accuracy
  • Multi-matrix compatibility for tissues, cells, DBS, feces, and plant extracts
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What Is the Mevalonate Pathway and Why Analyze It

The mevalonate (MVA) pathway is a central metabolic route that supplies precursors for isoprenoid biosynthesis in mammals, plants, and microorganisms. Together with the plastidial MEP/DOXP pathway in plants, it generates isoprenyl diphosphates that support essential cellular processes such as protein prenylation, membrane assembly, and stress adaptation.

These pathways give rise to biologically important lipid classes, including sterols (cholesterol in mammals, phytosterols in plants), ubiquinones (CoQ), plastoquinone, dolichols or polyprenols, and vitamin K compounds. Measuring these intermediates provides a direct readout of pathway activity.

Quantitative analysis of the mevalonate network helps reveal metabolic bottlenecks, lipid synthesis capacity, and responses to genetic or compound intervention. It is widely used to study mitochondrial function, redox balance, membrane biology, and growth regulation. Targeted LC–MS profiling enables precise measurement of pathway intermediates across diverse sample types for unbiased biological interpretation.

What Problems We Help You Solve

  • Dissect regulatory bottlenecks across MVA and MEP networks
    Quantifying key intermediates helps reveal control points in sterol and isoprenoid metabolism.
  • Connect pathway activity to genetic or compound perturbations
    Use defined metabolite shifts to validate enzyme targeting or expression-driven metabolic rewiring.
  • Resolve structurally related metabolites in complex matrices
    Confidently distinguish closely eluting or isobaric species across sterol, quinone, and prenol lipid classes.
  • Integrate plant and mammalian samples into a unified analytical framework
    Matrix-matched methods allow you to compare across species, tissues, and experimental systems.
  • Support studies in redox function, growth regulation, and membrane biology
    Quantitative profiles of pathway products link upstream biosynthesis to functional outputs in mitochondria, plastids, and ER.

How Mevalonate Pathway Analysis Supports Your Research

Reveal control points in isoprenoid metabolism

Quantify HMG-CoA, mevalonate derivatives, and prenyl diphosphates to locate pathway bottlenecks under compound treatment, nutrient shifts, or genetic manipulation.

Evaluate sterol biosynthesis in animal and plant systems

Track sterol intermediates such as lanosterol, zymosterol, desmosterol, sitosterol, and stigmasterol to assess flux, accumulation, or desaturation activity.

Understand redox-linked lipid pathways

Analyze CoQ (CoQ6–CoQ12), plastoquinone, and phylloquinone chain-length variants to explore mitochondrial and chloroplast redox potential.

Estimate prenylation and membrane-targeting potential

Measure farnesyl and geranylgeranyl diphosphates to gauge prenylation capacity and its role in signaling or membrane association.

Connect pathway data to broader lipidomics

Integrate targeted MVA/MEP outputs with untargeted lipidomics or redox panels to build system-level insight into lipid biosynthesis and organelle function.

Detectable Mevalonate Pathway Analyte List (Complete Panel)

Our panel includes key intermediates, diphosphates, sterol precursors, and lipid end-products from both the mevalonate (MVA) and MEP/DOXP pathways. Coverage spans mammalian and plant-specific metabolites.

Core Mevalonate Pathway Intermediates

Compound Superclass Class
Acetyl-CoA Lipids and lipid-like molecules Fatty acyls
Acetoacetyl-CoA Lipids and lipid-like molecules Fatty acyls
3-Hydroxy-3-methylglutaryl-CoA (HMG-CoA)
Mevalonic acid Lipids and lipid-like molecules Fatty acyls
Mevalonic acid 5-phosphate
Mevalonic acid 5-pyrophosphate Organic oxygen compounds Organic oxoanionic compounds

MEP Pathway Intermediates (Plant Only)

Compound Superclass Class
1-Deoxy-D-xylulose 5-phosphate (DXP) Organic oxygen compounds Sugar phosphates
Methylerythritol phosphate (MEP)
CDP-ME
CDP-ME2P
MEcPP
HMBPP

Prenyl Diphosphates (IPP, DMAPP, GPP, FPP, GGPP)

Compound Superclass Class
Isopentenyl pyrophosphate (IPP)
Dimethylallyl pyrophosphate (DMAPP) Lipids and lipid-like molecules Prenol lipids
Geranyl pyrophosphate (GPP) Lipids and lipid-like molecules Prenol lipids
Farnesyl pyrophosphate (FPP) Lipids and lipid-like molecules Prenol lipids
Geranylgeranyl pyrophosphate (GGPP) Lipids and lipid-like molecules Prenol lipids

Sterol Precursors and Cholesterol

Compound Superclass Class Source
Lanosterol Lipids and lipid-like molecules Prenol lipids Mammal (MVA)
Dihydrolanosterol Lipids and lipid-like molecules Prenol lipids Mammal
Zymosterol Lipids and lipid-like molecules Steroids and steroid derivatives Mammal
Zymostenol Lipids and lipid-like molecules Steroids and steroid derivatives Mammal
Lathostenol Mammal
7-Dehydrodesmosterol Lipids and lipid-like molecules Steroids and steroid derivatives Mammal
7-Dehydrocholesterol Lipids and lipid-like molecules Steroids and steroid derivatives Mammal
Desmosterol Lipids and lipid-like molecules Steroids and steroid derivatives Mammal
Cholesterol Lipids and lipid-like molecules Steroids and steroid derivatives Mammal
Cycloartenol Lipids and lipid-like molecules Steroids Plant (MVA)
24-Methylene cycloartanol Plant
Campesterol Steroids and steroid derivatives Plant
Sitosterol Steroids and steroid derivatives Plant
Stigmasterol Steroids and steroid derivatives Plant

Squalene and Oxidosqualene

Compound Superclass Class
Squalene Lipids and lipid-like molecules Prenol lipids
2,3-Oxidosqualene Lipids and lipid-like molecules Prenol lipids

Coenzyme Q, Plastoquinones, Dolichols, and Polyprenols

Compound Superclass Class Source
CoQ6–CoQ12 Both
Dolichols (13–21 isoprene units) Mammal
Polyprenols (varied chain length) Plant
Plastoquinone (PQ-9, PQ-10) Plant
Phylloquinone (Vitamin K1) Plant
Menaquinones (Vitamin K2 variants) Mammal

Why Choose Our Mevalonate Pathway Analysis Service?

  • Quantitative Accuracy

Standard curves with R2 ≥ 0.995 ensure reliable linearity across concentration ranges.

  • Reproducibility

Intra- and inter-batch precision maintained at CV ≤ 15% for qualified targets.

  • Sensitive Detection

Stabilized methods support low-abundance intermediates like phosphorylated species and dolichols.

  • High-Confidence Identification

Optional FTMS offers ≤ 5 ppm mass accuracy and MS2 validation for structural confirmation.

  • Low Background and Carryover

Signal carryover controlled under 1%; blanks confirm system cleanliness.

  • Multi-Matrix Compatibility

Protocols adapted for plasma, cells, tissues, feces, and DBS to support diverse study designs.

Which Methods Are Used in Mevalonate Pathway Profiling?

We apply advanced LC–MS instrumentation and method-specific optimizations to ensure accurate, reproducible quantification of mevalonate pathway metabolites. All analyses are performed under stringent quality control parameters.

UPLC–MRM/MS for Absolute Quantitation

Platform: Agilent 6495C quadrupole mass spectrometer

Mode: Scheduled MRM, polarity switching

Calibration: Multi-point external curve (R2 ≥ 0.995), with isotope-labeled or class-matched internal standards

Typical CVs: ≤ 15% for qualified targets

Use: Core quantitation of HMG-CoA, mevalonate derivatives, isoprenyl pyrophosphates, and sterols; includes support for MEP-derived intermediates in plant samples

UPLC–FTMS for Identity Confirmation (Optional)

Platform: Thermo Orbitrap series (e.g., Q Exactive, Exploris)

Mode: Full-scan HRAM with PRM or data-dependent MS2

Resolution: ≥ 60,000 at m/z 200; mass error ≤ 5 ppm

Mass accuracy: ≤ 5 ppm

Use: Structural verification of isobars (e.g., squalene vs. oxidosqualene), and resolution of CoQ, dolichol, plastoquinone, and phytosterol homologs

Chromatography Conditions

Columns: C18 or HILIC (2.1 × 100 mm, sub-2 μm particle size)

Mobile Phases: Aqueous ammonium formate or acetate (pH-controlled); acetonitrile or methanol as organic modifier

Flow Rate & Gradient: Optimized for analyte class; supports ≥10 points per peak in MRM

Matrix Compatibility: Suitable for polar (phosphates) and nonpolar (sterols, quinones) fractions from both mammalian and plant tissues
Agilent 6495C Triple Quadrupole

Agilent 6495C Triple Quadrupole (Figure from Agilent)

Agilent 1260 Infinity II HPLC

Agilent 1260 Infinity II HPLC (Fig from Agilent)

Thermo Fisher Q Exactive

Thermo Fisher Q Exactive (Figure from Thermo Fisher)

Mevalonate Pathway Analysis Workflow: Step by Step

1

Project Intake and Target Confirmation

We confirm analyte coverage, sample matrix, and reporting units. Optional consultation supports study design alignment.

2

Sample Preparation and Stabilization

Matrix-specific extraction methods are applied, including derivatization or cold-chain handling to protect labile phosphates and sterols.

3

UPLC–MS Analysis

Quantitative analysis is performed via UPLC–MRM/MS using internal standards and multi-point calibration. Optional UPLC–FTMS provides high-resolution confirmation.

4

Quality Control and Validation

Each batch includes blanks, QCs, and calibrators. Metrics such as linearity, CV%, recovery, and carryover are reviewed before release.

5

Data Delivery and Reporting

You receive calibrated concentration data, annotated chromatograms, and optional interpretation mapped to biological pathways.

Mevalonate Pathway Analysis Workflow

Sample Requirements for Mevalonate Pathway Profiling

Sample Type Min. Amount Required Notes
Animal tissues ≥ 50 mg wet weight Snap-frozen preferred. Avoid freeze-thaw.
Cultured cells ≥ 1×106 cells (pellet) Wash with PBS and snap-freeze.
Plasma/serum ≥ 100 μL EDTA or heparinized. Avoid hemolysis.
Feces (mouse/human) ≥ 50 mg Freeze immediately after collection.
Dried blood spots (DBS) 3–5 punches (3 mm) Use Whatman 903 or equivalent. Store dry with desiccant.
Microbial pellets ≥ 108 cells Harvest during mid-log phase for optimal metabolite content.
Plant tissues ≥ 100 mg fresh/frozen Flash-freeze in liquid nitrogen. Avoid enzymatic degradation.
Plant callus or cell cultures ≥ 50 mg Rapid harvest and freeze. Submit culture conditions if possible.
Lysates or extracts ≥ 50 μL or ≥ 50 μg protein Buffer composition must be disclosed; avoid detergents or chelators.

Shipping Tips:

All biological samples should be shipped on dry ice unless pre-dried (e.g., DBS). Label clearly, include sample manifest, and use leak-proof containers. For global shipments, ensure customs documentation matches declared contents.

What You Receive: Deliverables from Mevalonate Pathway Analysis

  • Absolute metabolite concentrations (nmol/mg or nmol/mL) in Excel/CSV
  • Annotated chromatograms with retention time and transitions (PDF)
  • Calibration curves and QC summary (R2, CV%, LOD/LOQ)
  • Method summary with instrumentation and workflow details
  • Optional MS/MS spectra for identity confirmation
  • Optional interpretive report linking results to pathway insights
Heatmap showing relative levels of sterol intermediates such as lanosterol, zymosterol, and cholesterol across experimental groups.

Sterol Biosynthesis Intermediates Heatmap

Calibration curve for quantitative LC–MS/MS showing linearity with R<sup>2</sup>, LOD, and LOQ thresholds marked.

Calibration Curve with R2 and LOD/LOQ

Multiple EIC traces overlaid to compare signal profiles of mevalonate metabolites in LC–MS runs.

Overlaid Extracted Ion Chromatograms (EICs)

Mass spectrum showing m/z peaks of a mevalonate pathway metabolite with isotopic pattern resolution.

Mass Spectrum of a Representative Analyte

Applications of Mevalonate Pathway Analysis in Research and Industry

Metabolic Engineering

Identify rate-limiting steps to optimize isoprenoid and sterol biosynthetic yields in microbial or plant systems.

Drug Mechanism Studies

Evaluate the metabolic impact of enzyme inhibitors or pathway modulators targeting HMG-CoA reductase or downstream enzymes.

Mitochondrial Function Research

Quantify CoQ intermediates to assess redox balance and electron transport efficiency.

Plant Lipidomics

Explore cross-talk between the MVA and MEP pathways in plastidial lipid biosynthesis and stress adaptation.

Prenylation and Signaling Studies

Measure prenyl diphosphates (FPP, GGPP) to understand protein modification and membrane association processes.

Comparative Lipid Metabolism

Profile sterol and quinone variations across species, tissues, or developmental stages to reveal evolutionary adaptations.

What does a mevalonate/MEP pathway analysis actually measure?

It quantifies precursor pools (e.g., HMG-CoA, mevalonate), prenyl diphosphates (IPP→GGPP), and downstream lipids (sterols, CoQ/dolichols; in plants also plastoquinone/phylloquinone), giving a direct readout of pathway activity and regulatory nodes.

How do you distinguish the mevalonate (MVA) pathway from the MEP pathway in plant samples?

By targeting diagnostic intermediates unique to MEP (DXP→MEcPP→HMBPP) alongside universal prenyl diphosphates, then comparing relative abundances and isotope/HRMS signatures to separate cytosolic MVA from plastid MEP contributions.

How is metabolite identity confirmed beyond MRM transitions?

High-resolution MS with PRM/data-dependent MS2 verifies exact mass, isotopic pattern, and fragments—critical for resolving isobars like squalene vs oxidosqualene and for chain-length variants in CoQ or dolichols.

What analytical choices improve sensitivity for labile phosphorylated intermediates?

Chemical derivatization and tailored LC phases enhance retention and peak shape for mevalonate phosphates and prenyl pyrophosphates, significantly boosting detection limits in complex matrices.

Can pathway profiling inform drug mechanism or target engagement?

Yes—altered levels of mevalonate intermediates and prenyl donors reveal on-pathway inhibition (e.g., HMG-CoA reductase) and can relate to biological effects from cholesterol synthesis to prenylation-dependent signaling and immunity.

What's the value of adding sterol intermediates to a cholesterol study?

Measuring lanosterol→desmosterol→cholesterol and related branches exposes flux control and compensatory routing, improving interpretation beyond total cholesterol alone.

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