Jasmonates (JAs) Analysis Service

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Overview

Jasmonates (JAs) are a class of phytohormones that mediate signaling pathways of plant defense responses. JAs are derivatives of fatty acids, including compounds such as jasmonic acid (JA), jasmonate isoleucine conjugate (JA-ILE), methyl jasmonate (MEJA), dihydrojasmonic acid (H2JA), and 12-oxo-phytodienoic acid (12-OPDA). Accurate detection and absolute quantification of JA and its conjugate(s) are essential to better understand the JA-mediated signaling and metabolic pathways. Traditionally, gas chromatography-mass spectrometry (GC-MS) is used for JA analysis. However, this method involves the separation of high boiling compounds and long chromatographic separation at elevated temperatures that may cause thermal decomposition of the molecules, increasing the difficulty of quantifying small amounts of JA accurately. Tandem mass spectrometry (LC-MS/MS) technology is also widely applied to measure JAs since it is a powerful tool for rapidly and accurately measuring small metabolites, such as JA and its amino acid conjugates. Creative Proteomics has developed a rapid method using LC-MS/MS for accurate jasmonate quantification in plants.

Figure 1. Derivatives and metabolites of JA (Zhai Q et al., 2017).

Applications of JA Analysis

Identification of JA transporters to elucidate JA transport mechanisms
Reveal functions of JAs and their mechanism of action
Understand JA signaling and metabolic pathways
Use of JAs in induced defenses against pests and pathogens
Enhance crop quality for human utility

Advantages of Our JA Analysis Service

Improved sample preprocessing offers high recovery rates and effectiveness

Enable measurement of JA, MEJA, H2JA, JAILE, and 12-OPDA

Accurate detection and quantification of JAs with LC-MS platforms

Customizable bioinformatics analysis

End-to-end JA analysis with comparable prices

Service Workflow

Our newly developed sample preprocessing method provides safety, environmental protection, rapidness, effectiveness, and recovery rates consistently exceeding 85%. Sample preprocessing at Creative Proteomics complies with in-house standard operation procedures (SOPs), ensuring maximized homogeneity between samples.

Service Workflow

Quantification methods: external reference method or isotope-labeled internal standard method

Mode: MRM, capable of simultaneously detecting more than 1000 MRM ion pairs

Precision: ≤10^-9 g

Positive/Negative polarity switching time: ≤20 ms, allowing for the acquisition of Q1/Q3 MRM transition mass spectra in both ionization modes from a single LC-MS/MS run.

Analysis content:

Standard curve creation
Raw data preprocessing
Absolute quantification of JAs
Differential metabolites screening
Optimal analyses such as KEGG pathway analysis and hierarchical clustering

List of Detectable Jasmonates at Creative Proteomics

Detectable Jasmonates	CAS	Quantification Methods
JA	3572-66-5	External reference method / isotope-labeled internal standard method
H2JA	3572-64-3	External reference method / isotope-labeled internal standard method
JA-ILE	120330-93-0	External reference method / isotope-labeled internal standard method
12-OPDA	85551-10-6	External reference method / isotope-labeled internal standard method
EMJA	879490-89-8	External reference method / isotope-labeled internal standard method

Sample Requirements

1. Fresh plant tissues from leaf, flower, stem, root, or fruit: >2 g. Provide young plant tissues for best results. Samples should be frozen in liquid nitrogen immediately after collection, and then transferred to -80°C for storage.

2. Plant seeds: >2 g.

At least 3 biological replicates.

Deliverables

Experimental procedure
Parameters of liquid chromatography and MS
MS raw data files and MS data quality checks
JA quantification data
Custom analysis report

Creative Proteomics provides plant hormones analysis services utilizing LC-MS/MS technology for accurate identification and quantitation of phytohormones and revelation of the roles of these molecules in plant regulation and stress responses. Please do not hesitate to contact us for detailed information about the jasmonate analysis service.

References

Tamogami S, Agrawal G K, Rakwal R. Targeted Quantitative Analysis of Jasmonic Acid (JA) and Its Amino Acid Conjugates in Plant Using HPLC-Electrospray Ionization-Tandem Mass Spectrometry (ESI-LC-MS/MS). Sample Preparation in Biological Mass Spectrometry. Springer, Dordrecht, 2011: 869-875.
Zhai Q, Yan C, Li L, et al. Jasmonates. Hormone Metabolism and Signaling in Plants, 1st ed.; Li, J., Li, C., Smith, S., Eds, 2017: 243-271.
Rohwer C L, Erwin J E. Horticultural applications of jasmonates. The Journal of Horticultural Science and Biotechnology, 2008, 83(3): 283-304.