Organic Acid and Metabolites Analysis
Service Details Case Study

What are Organic Acids?

Organic acids are a class of compounds characterized by the presence of one or more carboxyl groups (COOH) attached to a carbon atom. These acids can be classified as either aliphatic or aromatic based on their carbon backbone structure. Examples of aliphatic organic acids include acetic acid, citric acid, and lactic acid, while benzoic acid and salicylic acid are examples of aromatic organic acids. Organic acids are ubiquitous in nature and can be found in various sources, including plants, animals, and microorganisms.

Carbohydrate metabolism refers to the biochemical processes by which carbohydrates are synthesized, broken down, and converted into usable forms of energy. This metabolic pathway involves a series of enzymatic reactions that occur within cells. Understanding carbohydrate metabolism is critical for comprehending the underlying mechanisms of energy production, glycosylation, and the regulation of various cellular processes.

Organic Acid Metabolism

Organic acid metabolism refers to the biochemical processes that involve the synthesis, degradation, and interconversion of organic acids within living organisms. These processes are tightly regulated and are essential for maintaining cellular homeostasis. Organic acids are involved in a wide range of metabolic pathways, including energy production, amino acid metabolism, and the tricarboxylic acid (TCA) cycle.

Analyzing organic acid metabolism provides valuable insights into the functioning of different cellular processes. Abnormal levels of specific organic acids can indicate underlying metabolic disorders or diseases. By identifying and quantifying these organic acids, researchers and clinicians can better understand the metabolic status of an organism and develop targeted interventions for various conditions.

Organic Acid Metabolomics Service Provided by Creative Proteomics

Services Offered by Creative Proteomics

Targeted Organic Acid Profiling: Creative Proteomics offers targeted analysis of specific organic acid metabolites of interest. By employing highly sensitive mass spectrometry techniques, researchers can accurately quantify the levels of targeted organic acids. This service allows for a focused investigation into the role and regulation of specific organic acids in cellular metabolism.

Untargeted Metabolomics: Creative Proteomics also provides untargeted metabolomics analysis, which enables a comprehensive assessment of the entire organic acid metabolome. Using cutting-edge mass spectrometry platforms, this approach allows for the detection and identification of a wide range of organic acids, including both known and unknown metabolites. Untargeted metabolomics provides a global view of organic acid metabolism, uncovering novel insights and potential biomarkers.

Analytical Techniques Employed by Creative Proteomics

To ensure accurate and reliable results, Creative Proteomics utilizes state-of-the-art analytical techniques for organic acid metabolism analysis:

Liquid Chromatography-Mass Spectrometry (LC-MS): Creative Proteomics employs liquid chromatography-mass spectrometry (LC-MS) for organic acid analysis. LC-MS combines the separation capabilities of liquid chromatography with the sensitive and selective detection of mass spectrometry. This powerful technique allows for the identification and quantification of organic acids with high precision. Creative Proteomics employs advanced LC-MS systems such as the Agilent 1290 Infinity LC System coupled with the Agilent 6530 Accurate-Mass Q-TOF LC/MS System to perform organic acid metabolism analysis.

Gas Chromatography-Mass Spectrometry (GC-MS): For volatile organic acids, Creative Proteomics utilizes gas chromatography-mass spectrometry (GC-MS). This technique involves the separation of analytes using gas chromatography and subsequent detection and quantification by mass spectrometry. The Shimadzu GCMS-QP2020 NX Gas Chromatograph Mass Spectrometer is one of the sophisticated instruments employed by Creative Proteomics for GC-MS analysis in organic acid metabolism studies.

Workflow for Plant Metabolomics ServiceWorkflow for Plant Metabolomics Service

List of Organic Acid Metabolites Analyzed (including but not limited to)

Category Organic Acid
Ketone Metabolites Acetone
Detoxification α-Hydroxybutyrate, β-Hydroxybutyrate, Butyrate, α-Ketovaleric acid, α-Keto-β-methylvaleric acid, β-Hydroxy-β-methylbutyrate, α-Keto-isocaproic acid, Methylmalonic acid, Glutaric acid, Hydroquinone, Lactate, Formate, Benzoic acid, Hippuric acid
TCA Cycle Malic acid, Citrate, Isocitrate, α-Ketoglutarate, Succinate, Fumarate, Malate, Oxaloacetate
Glycolysis Pyruvate, Lactic acid
Cofactor Requirement Propionate, Hexanedioic acid (Adipic acid), Ethylmalonic acid, Methylsuccinic acid, Cinnamic acid, Salicylic acid, Ursolic acid, Ellagic acid, Ferulic acid, Chlorogenic acid, Maleic acid, Glutamic acid, Glycolic acid, Glyoxylic acid, 4-Hydroxybenzoic acid, 4-Hydroxycinnamic acid, 4-Hydroxyphenylacetic acid, 4-Hydroxyphenylpyruvic acid
Neurotransmitter Metabolism Tryptophan metabolites (Kynurenic acid, Quinolinic acid, 5-Hydroxyindoleacetic acid, Nicotinic acid)
Microbial Metabolism Methylparaben, Phenol, Catechol, Ethyl acetate, Tri-carboxylic acid

Why Need Organic Acid Analysis?

Metabolic Profiling: Organic acid metabolism analysis allows for the comprehensive profiling of metabolic pathways in organisms. By identifying and quantifying organic acids, researchers can gain a deeper understanding of the interconnectedness of metabolic reactions, the flow of metabolites, and the regulation of metabolic pathways.

Stress Responses and Environmental Adaptation: By studying changes in organic acid profiles under various stress conditions (e.g., drought, temperature extremes, nutrient deficiencies), researchers can unravel the mechanisms by which plants respond and adapt to their surroundings.

Plant-Microbe Interactions: Organic acids play a crucial role in plant-microbe interactions, including symbiotic relationships with beneficial microbes and defense responses against pathogens. Analyzing organic acid metabolism can provide insights into the dynamic interplay between plants and microorganisms, shedding light on the mechanisms of disease resistance, nutrient uptake, and overall plant health.

Secondary Metabolism and Phytochemical Production: Many organic acids are precursors or intermediates in the biosynthesis of secondary metabolites, including phenolics, flavonoids, and terpenoids, which contribute to the nutritional and medicinal properties of plants. Analyzing organic acid metabolism allows researchers to understand the biosynthetic pathways and regulatory mechanisms involved in phytochemical production, facilitating efforts to enhance the nutritional value and therapeutic potential of crops.

Biomarker Discovery: Alterations in organic acid profiles can indicate metabolic dysregulation, disease conditions, or responses to external stimuli. By analyzing organic acid metabolism, researchers can identify specific biomarkers that can be used for diagnostic, prognostic, or therapeutic purposes in fields such as clinical medicine, agriculture, and environmental monitoring.

Sample Requirements for Organic Acid Metabolites Assay

Sample Types Minimum Sample Size Biological Repeat
Plant Samples Roots, stems and leaves, floral parts, fruits/seeds, rhizomes, buds/tender leaves, tissue sections, pollen, bark, trunk/wood, resin/gum, resin acids, seedlings/young plants, rhizosphere soil, root exudates. 100 mg - 1 g 3-6
Animal Samples Blood/plasma
Urine
Tissues (liver, muscle, adipose, etc.), brain tissue
Feces
1-2 ml
5-10 ml
100-200 mg
1-2 g
Humans >30/group

Animals 8-10/group

Cell Samples Adherent cells, suspension cells, cell cultures 1×107 Humans >30/group
Animals 8-10/group

Case 1. GC-MS Analysis Reveals Metabolic Changes in Organic Acids during Drought Stress in Plants

Background:

Metabolic changes in organic acids play a crucial role in plant responses to drought stress. Organic acids are important metabolic products in plant cells, and their alterations can reflect the plant's adaptation to environmental stress.

Technical Methodology:

The analysis of organic acid metabolism was performed using GC-MS (Gas Chromatography-Mass Spectrometry) techniques. This method allows for the identification and quantification of organic acids present in the samples.

The workflow for the analysis involved the following steps:

  • Sample preparation: Plant tissues were collected and processed to extract organic acids.
  • Derivatization: The extracted organic acids were derivatized to enhance their detectability in the GC-MS system.
  • GC-MS analysis: The derivatized organic acids were injected into the GC-MS system, which separates and detects the compounds based on their physicochemical properties.
  • Data analysis: The obtained chromatograms and mass spectra were analyzed using appropriate software to identify and quantify the organic acids present in the samples.

Results

The results of the analysis revealed significant changes in the content of various organic acids under drought stress conditions. Specifically, succinic acid, malic acid, and galacturonic acid showed the highest increases in response to long-term drought stress. Other organic acids, such as methylmalonic acid, citric acid, glyceric acid, and isocitric acid, exhibited a decrease in content compared to the control samples.

The study also mentioned the presence of phenolic acids, including caffeic acid and ferulic acid, in cereal grains such as wheat, oat, barley, and rye. These phenolic acids showed elevated levels, suggesting their potential role in drought stress responses.

Overall, the analysis of organic acid metabolism provided insights into the metabolic changes occurring in plants under drought stress conditions. These findings contribute to a better understanding of plant responses to environmental stress and the potential mechanisms involved in drought tolerance.

Key mechanism of stress avoidance targeted by humic acid and fulvic acidKey mechanism of stress avoidance targeted by humic acid and fulvic acid (Van Oosten et al., 2017)

Reference

  1. Khan, Naeem, et al. "Role of sugars, amino acids and organic acids in improving plant abiotic stress tolerance." Pak. J. Bot 52.2 (2020): 355-363.
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