Plant Hormones Analysis Service: Precision Quantification for Phytohormone Profiling
When plant hormone levels shift, it signals fundamental changes in growth regulation, stress adaptation, and developmental programming. Creative Proteomics provides high-sensitivity, targeted LC–MS/MS and GC–MS plant hormones analysis to quantify classic, emerging, and conjugated phytohormones across diverse plant tissues and matrices.
Our phytohormone profiling service is engineered for plant scientists, crop breeders, and agricultural biotechnologists who require absolute, isomer-aware quantification rather than simple qualitative detection.
- Resolve complex isomers (e.g., cis/trans zeatins) with optimized chromatography.
- Absolute quantification using stable isotope-labeled internal standards.
- Deep coverage from trace-level strigolactones to high-abundance abscisic acid.
- High-throughput workflows designed for large-scale mutant screens and field trials.
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- What We Provide
- Analytes
- Advantages
- Technology Platform
- Sample Requirements
- Demo
- Case Study
- FAQs
Biological Roles of Plant Hormones and Why Their Analysis Matters
Plant hormones coordinate growth and development (cell division/elongation, organ patterning, flowering, fruiting) and orchestrate responses to environmental and biotic stress (drought, salinity, temperature, pathogens, herbivory). Because hormone pathways are interconnected—often through biosynthesis, conjugation, transport, and catabolism—a multi-class panel is frequently needed to interpret phenotype and pathway-level regulation.
Targeted profiling helps you:
- Identify which hormone pathways shift under stress or genetic perturbation
- Track bioactive forms vs. storage/conjugated forms
- Evaluate cross-talk among auxins, cytokinins, gibberellins, ABA, jasmonates, salicylates, brassinosteroids, and strigolactones
Plant Hormones Quantification & Profiling Services
You can select from or combine the following offerings:
- Targeted phytohormone quantification
Absolute or relative quantitation of key hormones and metabolites in plant tissues and related matrices. - Custom hormone panels
Panel configuration around your hypothesis (e.g., drought/ABA axis, defense JA–SA balance, growth auxin–CK–GA modules). - Multi-tissue and time-course profiling
Harmonized workflows for cross-tissue comparisons and developmental or treatment time series. - Optional statistics and pathway summaries
Group comparisons and visualization-ready outputs for quick biological interpretation.
Service Selection Guide: Matching Your Research Goals to the Right Solution
| Your Research Objective | Recommended Technical Solution | Core Advantages |
|---|---|---|
| Fundamental Physiological Mechanisms (e.g., studying gene-hormone interactions, developmental stages) | Targeted Absolute Quantification (LC-MS/MS) | Gold Standard Data: Absolute concentrations with internal standards. High sensitivity for defining precise hormonal shifts. |
| Large-Scale Screening (e.g., mutant libraries, GWAS populations, field trials) | High-Throughput Phytohormone Profiling | Cost-Effective & Fast: Simultaneous semi-quantification of major hormone classes suitable for identifying outliers in large sample sets. |
| Ultra-Trace or Novel Hormone Analysis (e.g., Strigolactones, Brassinosteroids, or unique conjugates) | Custom Method Development & Validation | Tailored Protocols: Specialized extraction and enrichment strategies designed to overcome complex matrices and push detection limits. |
Comprehensive Detectable Plant Hormones & Metabolites
Service Contents
- Auxins (Indolic and Non-Indolic)
- Cytokinins (Adenine-type and Phenylurea-type)
- Gibberellins (GAs)
- Abscisic Acid (ABA) and Related Compounds
- Jasmonates (JAs) and Oxylipins
- Salicylates and Phenolics
- Brassinosteroids (BRs)
- Strigolactones (SLs) and Others
Auxins (Indolic and Non-Indolic)
- Indole-3-acetic acid (IAA)
- Indole-3-butyric acid (IBA)
- Indole-3-propionic acid (IPA)
- 4-Chloroindole-3-acetic acid (4-Cl-IAA)
- Phenylacetic acid (PAA)
- Indole-3-acetyl-aspartic acid (IAA-Asp)
- Indole-3-acetyl-glutamic acid (IAA-Glu)
- Methyl indole-3-acetate (Me-IAA)
Cytokinins (Adenine-type and Phenylurea-type)
- Isopentenyladenine (iP) types: iP, iPR, iP7G, iP9G, iPRMP
- trans-Zeatin (tZ) types: tZ, tZR, tZ7G, tZ9G, tZOG, tZROG, tZRMP
- cis-Zeatin (cZ) types: cZ, cZR, cZ9G, cZOG, cZROG
- Dihydrozeatin (DZ) types: DZ, DZR, DZ9G, DZOG, DZROG
- Aromatic CKs: Benzyladenine (BA), Kinetin (K), Topolin isomers (oT, mT, pT)
- Bioactive GAs: GA1, GA3, GA4, GA7
- Precursors/Intermediates: GA9, GA12, GA15, GA19, GA20, GA24, GA44, GA53
- Inactivated Metabolites: GA8, GA29, GA34, GA51
Abscisic Acid (ABA) and Related Compounds
- (+)-Abscisic acid (ABA)
- ABA-glucose ester (ABA-GE)
- Phaseic acid (PA)
- Dihydrophaseic acid (DPA)
- 7′-Hydroxy-ABA
- Neophaseic acid (neoPA)
Jasmonates (JAs) and Oxylipins
- Jasmonic acid (JA)
- Jasmonoyl-isoleucine (JA-Ile)
- Methyl jasmonate (MeJA)
- 12-Oxo-phytodienoic acid (12-OPDA)
- Dihydro-JA
- 7-iso-Jasmonic acid
Salicylates and Phenolics
- Salicylic acid (SA)
- Salicylic acid 2-O-β-D-glucoside (SAG)
- Methyl salicylate (MeSA)
- Brassinolide (BL)
- Castasterone (CS)
- Typhasterol (TY)
- 6-Deoxocastasterone
- 24-Epibrassinolide
Strigolactones (SLs) and Others
- 5-Deoxystrigol
- Strigol, Sorgomol, Orobanchol
- Ethylene precursor: 1-Aminocyclopropane-1-carboxylic acid (ACC)
- Others: Melatonin; N-Acetylserotonin
Why Choose Our Plant Hormones Analysis
Selecting a phytohormone analysis partner requires confidence in sensitivity, quantitative accuracy, and biological relevance. Our key advantages include:
- Attomole-Level Sensitivity
Optimized positive- and negative-ion ESI enables limits of detection down to 0.01–0.5 pg/mg fresh weight, supporting ultra-low-abundance targets such as strigolactones. - High Recovery for Labile Analytes
A proprietary double-extraction workflow with pre-chilled solvents (−20°C) and antioxidants (e.g., BHT) delivers 85–95% recovery, protecting unstable compounds such as JA-Ile and GA4. - Isotope-Dilution Absolute Quantitation
50+ stable isotope-labeled internal standards (13C, 2H, 15N) correct matrix effects in every sample, achieving CV < 12%. - Isomer-Resolved Chromatography
Sub-2 µm UPLC columns and ~25 min gradients provide baseline separation of critical isomers, including trans- vs. cis-zeatin. - Validated Across Difficult Matrices
Proven protocols for woody tissues, recalcitrant seeds, siliques, and specialized fluids such as xylem/phloem sap.
Plant Hormones Analysis Workflow: Step-by-Step Process
Instrumentation and Methods for Plant Hormones Profiling
To provide the highest levels of sensitivity and reproducibility, Creative Proteomics utilizes a multi-platform approach. We combine the absolute quantification power of Triple Quadrupole (QqQ) systems with the high-resolution discovery capabilities of Orbitrap technology.
Advanced Analytical Platforms for Phytohormone Profiling
| Platform | Core Instrumentation | Key Capabilities | Technical Highlights |
|---|---|---|---|
| Targeted LC-MS/MS | Agilent 1290 Infinity II + 6495C Triple Quad (QqQ) | Absolute Quantification via MRM mode. The "Gold Standard" for phytohormone flux analysis. | Attomole-level sensitivity; iFunnel technology for trace-level signal enhancement; wide dynamic range. |
| High-Res LC-MS (HRAM) | Orbitrap Exploris 480 / Q Exactive HF-X | Non-targeted Profiling and isomer resolution. Ideal for discovery and complex conjugates. | 480,000 FWHM resolution;<1 ppm mass accuracy; eliminates near-isobaric interference from plant matrix. |
| GC-MS | Agilent 7890B GC + 5977B MSD | Volatile & Small Molecule Analysis. Specialized for MeJA, MeSA, and precursors. | High-efficiency Electron Ionization (EI); robust separation of derivatized hormone isomers. |
Methodological Optimization
Our methods are refined to overcome the specific challenges of plant biochemistry:
- Isomer-Specific Separation: Optimized UPLC gradients baseline-resolve bioactive hormones from their inactive isomers (e.g., t-Zeatin vs. c-Zeatin).
- Isotope Dilution: Every run is spiked with stable isotope-labeled standards ($^{13}C$ or $^{2}H$) to correct for matrix-induced signal suppression.
- Ultrapure Extraction: Multi-step Solid Phase Extraction (SPE) ensures the removal of chlorophyll, tannins, and lipids that can compromise instrument performance.
Agilent 1290 Infinity II HPLC (Figure from Agilent)
Agilent 6495C Triple Quadrupole (Figure from Agilent)
Thermo Fisher Q Exactive (Figure from Thermo Fisher)
Agilent 7890B-5977A (Figure from Agilent)
Sample Requirements for Plant Hormones Analysis
| Item | Recommendation / Requirement |
|---|---|
| Accepted sample types | Fresh or frozen plant tissues (leaf, root, stem, flower, seed, silique), callus, culture media; specialized fluids (xylem/phloem sap) |
| Minimum amount (typical) | 50–200 mg fresh weight per sample for most tissues; 200–500 mg recommended for very low-abundance targets (e.g., strigolactones) |
| Replicates | ≥ 3 biological replicates per group; technical replicates optional |
| Quenching (recommended) | Flash-freeze in liquid nitrogen immediately after harvest to minimize metabolic changes |
| Homogenization | Grind to a fine powder under cryogenic conditions (liquid nitrogen) |
| Storage | −80°C (preferred) or −20°C (short term); avoid repeated freeze–thaw cycles |
| Packaging | Pre-labeled screw-cap tubes (leak-proof, O-ring recommended); include blanks/controls if available |
| Shipment | Ship on dry ice; samples must remain frozen throughout transit |
Demo Results
Calibration curve for targeted phytohormone quantitation showing wide linear range and low LOD/LOQ by LC–MS/MS.
Isotope-dilution internal standards reduce matrix effects and improve precision (lower CV) in plant hormone analysis.
Extracted ion chromatograms demonstrate baseline separation of key isomers (trans- vs cis-zeatin) for reliable calls.
Heatmap summarizes multi-class phytohormone shifts across groups, supporting biological interpretation of treatment effects.
Case Study
Targeted phytohormone LC–MS/MS reveals distinct hormone signatures in agro-industrial byproducts to support application-specific sorting
Journal: Bioresources and Bioprocessing
Published: 2024
- Background
- Analytical Approach
- Key Findings
- Takeaways for Project Design
Agro-industrial byproducts and wastes are increasingly explored as sustainable inputs for agriculture and bioprocessing. However, their biological effects can vary widely by source and batch. In this study, researchers asked whether phytohormone composition could help characterize and differentiate byproduct streams, enabling more informed sorting and downstream use.
At study endpoints, selected agro-industrial byproducts (including wheat bran and garlic processing residues) were profiled using a targeted phytohormone LC–MS/MS workflow performed by Creative Proteomics. Approximately 25 mg of dry material was extracted for analysis. The panel covered key hormone classes and regulators, including ABA, SA, JA, JA-Ile, OPDA, IAA and IAA conjugates, cytokinins (cZ/tZ and ribosides), strigolactones (strigol, orobanchol), and GA precursors (GA12/GA19/GA53). Quantitation was normalized using stable isotope internal standards (e.g., D6ABA, D5IAA, D4SA, D2JA, D2GA1), and separation was performed on a C18 column (ZORBAX Eclipse Plus, 2.1 × 100 mm).
The targeted dataset showed that different byproduct streams carried distinct phytohormone profiles rather than a uniform "plant extract" background. Several bioactive hormones and metabolites were detected across samples, while specific analytes fell below quantification limits in all tested materials (including multiple bioactive GAs). Notably, the study reported high strigolactone levels in one byproduct stream and orobanchol enrichment in another, alongside measurable levels of IAA, SA, ABA, jasmonates, cytokinins, and GA precursors, supporting the idea that hormone signatures can inform how these materials may influence plant growth and development.
This case highlights how a focused, targeted LC–MS/MS phytohormone panel can:
- Differentiate heterogeneous plant-derived materials using quantitative hormone fingerprints
- Identify bioactive vs. low/undetectable hormone classes to guide realistic biological expectations
- Support application decisions (e.g., biostimulant screening, agronomic testing) with mechanism-relevant chemical evidence
FAQ of Plant Hormone Analysis
What are the lowest limits of quantification (LLOQ) for various hormones analyzed by Creative Proteomics?
| Compound | LOD, nM 10 uL injection |
|---|---|
| ABA | 0.8000 |
| cZ | 0.0128 |
| cZR | 0.0128 |
| IAA | 0.8000 |
| IAA-Ala | 8.0000 |
| IAA-Asp | 0.4000 |
| IAA-Trp | 0.4000 |
| JA | 0.3200 |
| JA-ILE | 0.0800 |
| MethylIAA | 0.1600 |
| OPDA | 8.0000 |
| SA | 10.0000 |
| tZ | 0.0320 |
| tZR | 0.0128 |
| GA1 | 0.8000 |
| GA3 | 0.8000 |
| GA4 | 4.0000 |
| GA8 | 0.8000 |
| GA9 | 4.0000 |
| GA12 | 4.0000 |
| GA19 | 0.8000 |
| GA20 | 4.0000 |
| GA24 | 4.0000 |
| GA29 | 4.0000 |
| GA53 | 0.8000 |
Which analytical method is best for quantifying multiple plant hormone classes?
Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) with Multiple Reaction Monitoring (MRM) is the established standard for multi-class profiling. Unlike GC-MS, LC-MS/MS does not require high-temperature derivatization, which is critical for preserving thermally labile and polar compounds such as gibberellins and cytokinins. For exploratory studies, high-resolution Orbitrap systems provide the mass accuracy needed to distinguish hormones from isobaric secondary metabolites.
How do you address matrix effects in complex tissues like woody stems or oily seeds?
Matrix effects, such as ion suppression from plant pigments and tannins, are mitigated through isotope-dilution internal standards. By spiking every sample with stable isotope-labeled analogs like 13C or 2H, we compensate for signal variance during electrospray ionization. Additionally, specialized solid-phase extraction (SPE) protocols remove interfering polyphenols and chlorophyll, ensuring high signal-to-noise ratios even in recalcitrant matrices.
What is the most critical factor for preserving phytohormone integrity during sampling?
Immediate metabolic quenching is essential. Plant hormones respond to harvesting stress within seconds, leading to rapid enzymatic turnover. Samples must be flash-frozen in liquid nitrogen immediately upon collection. Maintaining a strict cold-chain during cryogenic grinding and using chilled extraction solvents at -20°C prevents the artificial induction of stress hormones like jasmonic acid and the degradation of unstable molecules like indole-3-acetic acid.
Can phytohormones be detected in micro-samples like shoot apices or pollen?
Yes. Modern mass spectrometry platforms with enhanced ion sampling technology push detection limits to the attomole or low picogram range. This sensitivity enables the quantification of ultra-trace hormones like strigolactones or localized signals in milligram-scale tissues. We employ micro-extraction techniques that minimize sample loss, providing reliable data from localized tissues where hormone concentrations are sub-picomolar.
Why is baseline isomer resolution necessary for reliable hormone quantification?
Structural isomers often possess vastly different biological activities. For instance, trans-zeatin is highly active while cis-zeatin is typically associated with stress or dormancy. We use sub-2 μm UPLC columns and optimized gradients to achieve baseline resolution (Rs > 1.5) of these isomers. This ensures quantification is specific to the bioactive form, preventing the overestimation of hormone levels that occurs when isomers co-elute.
How can hormonal crosstalk be interpreted from quantitative analysis results?
Bioinformatics workflows integrate multi-class hormone data into principal component analysis (PCA) and fold-change heatmaps to visualize regulatory shifts. By mapping quantitative data onto metabolic pathways, researchers can identify key interaction hubs, such as the antagonism between abscisic acid and gibberellins during germination or the trade-offs in salicylic acid-jasmonic acid defense signaling.
Learn about other Q&A.
Publications
Here are some of the metabolomics-related papers published by our clients:
- Detailed analysis of agro-industrial byproducts/wastes to enable efficient sorting for various agro-industrial applications. Bioresources and Bioprocessing, 2024. https://doi.org/10.1186/s40643-024-00763-7
- A Water Solution from the Seeds, Seedlings and Young Plants of the Corn Cockle (Agrostemma githago) Showed Plant-Growth Regulator Efficiency. Plants, 2025. https://doi.org/10.3390/plants14152349
- WI12 Rhg1 interacts with DELLAs and mediates soybean cyst nematode resistance through hormone pathways. Plant Biotechnology Journal, 2022. https://doi.org/10.1111/pbi.13709
- Characterization of CYCLOPHILLIN38 shows that a photosynthesis-derived systemic signal controls lateral root emergence. Plant Physiology, 2021. https://doi.org/10.1093/plphys/kiaa032
- Overexpression of maize ZmLOX6 in Arabidopsis thaliana enhances damage-induced pentyl leaf volatile emissions that affect plant growth and interaction with aphids. Journal of Experimental Botany, 2023. https://doi.org/10.1093/jxb/erac522
