Metabolomics Creative Proteomics

Creative Proteomics Metabolomics

Salicylic Acid (SA) Analysis Service

Salicylic Acid (SA) Analysis Service

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Creative Proteomics offers salicylic acid (SA) analysis service to analyze SA and its derivatives based on state-of-the-art LC-MS/MS platforms in MRM mode, as well as tailored statistical analyses and bioinformatics analyses to meet your project needs. We are committed to putting our clients first, providing excellent and feasible services and achieving the best outcomes we can. Our complete sample-to-data services can help better understand the biosynthesis, function, metabolism, and mechanism of action of SA.

Overview

Plants have developed complex mechanisms to protect themselves from pathogen attacks and other environmental stresses. In recent years, salicylic acid (SA; 2-hydroxybenzoic acid) has emerged as an important plant hormone with key roles in the defense against biotrophic pathogens. Studies have identified SA receptor proteins and revealed the regulation of pathogen-induced SA biosynthesis and how SA regulates defense gene expression. SA is required for the expression of many defense genes and the assembly of root microbiome. Methyl salicylate (MeSA), a derivative of SA, acts as a long-distance mobile signal for inducing systemic acquired resistance (SAR) in uninfected parts of plants. SA is methylated by SA carboxyl methyltransferase (SAMT) to form MESA, which travels to the systemic tissues and activates resistance following its conversion back SA by SA-binding protein 2 (SABP2). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is generally used for the detection of SA. Creative Proteomics has developed an accurate and feasible protocol for the quantitative determination of salicylic acid in given plant tissues. Multiple reaction monitoring (MRM) is utilized for data acquisition, which is a highly specific and sensitive label-free technique for detecting and quantifying selected protein/peptides of interest.

SA undergoes many modifications in plantsFigure 1. SA undergoes many modifications in plants (Dempsey & Klessig 2017).

Applications of SA Analysis

  • Elucidate the regulation of SA biosynthesis and SA perception
  • Reveal the role of SA and MESA and their signal transduction mechanism
  • Clarify the relationships among SA, MESA, and other derivatives of SA
  • Use of SA in induced defenses against pathogens
  • Enhance crop quality for human utility

Advantages of Our SA Analysis Service

Improved sample preprocessing offers high recovery rates and Effectiveness

Accurate detection and quantification of SA and MESA

Flexibility to customize protocol and bioinformatics analyses to meet your specific project needs

End-to-end SA analysis with fast turnaround time and comparable price

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. Electrospray ionization (ESI) is performed with a triple quadrupole LC/MS system, and MRM is utilized for data acquisition.

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 SA and MESA
  • Differential metabolites screening
  • Optimal analyses such as KEGG pathway analysis and hierarchical clustering

List of Detectable SA at Creative Proteomics

Detectable SAsCASQuantification Methods
SA69-72-7External reference method / isotope-labeled internal standard method
MESA119-36-8External 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
  • SA and MESA quantification data
  • Custom analysis report

Creative Proteomics provides plant hormones analysis services to determine the level of specific hormones in plant tissues powered by state-of-the-art mass spectrometry coupled with gas chromatography (GC) or liquid chromatography (LC), which is essential for revealing the roles and functions of various phytohormones in plant growth, development, and resistance. Please do not hesitate to contact us for detailed information about our SA and MESA analysis services.

References

  1. Li Y, Zhang W, Dong H, et al. Salicylic acid in Populus tomentosa is a remote signalling molecule induced by Botryosphaeria dothidea infection. Scientific reports, 2018, 8(1): 1-9.
  2. Zhang Y, Li X. Salicylic acid: biosynthesis, perception, and contributions to plant immunity. Current opinion in plant biology, 2019, 50: 29-36.
  3. Dempsey D M A, Klessig D F. How does the multifaceted plant hormone salicylic acid combat disease in plants and are similar mechanisms utilized in humans?. BMC biology, 2017, 15(1): 1-11.
  4. Zheng W, Yoo K H, Abd El-Aty A M, et al. Quantitative determination of carbasalate calcium derived metabolites, acetylsalicylic acid and salicylic acid, in six animal foods using liquid-liquid extraction method coupled with liquid chromatography-tandem mass spectrometry. Food chemistry, 2019, 278: 744-750.
For Research Use Only. Not for use in diagnostic procedures.

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