Organic acids are acidic organic compounds (excluding amino acids) containing carboxyl groups that are widely found in living organisms. In animals and plants except a few organic acids exist in the free state, generally in the form of salt or ester combined with potassium, sodium, calcium plasma.
Many organic acids play an important role by directly participating in biochemical reactions in the process of life activities. For example: 1) antibacterial, antiviral and anticancer effects; 2) softening blood vessels and promoting the absorption of calcium and iron; 3) acting as intermediate products of metabolism; 4) helping gastric juice to digest fats and proteins; 5) Preventing diseases and promoting metabolic effects, etc. In food research, the accumulation of organic acids is an important indicator of the flavor quality of fruits.
Creative Proteomics has established a quantitative analysis method for organic acids based on LC-MS/MS platform, which simultaneously detects 60+ organic acids which can be applied to practical sample detection in metabolomics and foodomics.
Detectable Organic Acids Include but Not Limited to
| No. | Organic Acid | Metabolic Pathway |
|---|
| 1 | Acetic Acid | Citric Acid Cycle |
| 2 | Aconitic Acid | Citric Acid Cycle |
| 3 | Adipic Acid | Fatty Acid Metabolism |
| 4 | Ascorbic Acid (Vitamin C) | Various |
| 5 | Benzoic Acid | Phenylalanine Metabolism |
| 6 | Butyric Acid | Butyrate Production |
| 7 | Citric Acid | Citric Acid Cycle |
| 8 | Formic Acid | Various |
| 9 | Fumaric Acid | Citric Acid Cycle |
| 10 | Gluconic Acid | Gluconeogenesis |
| 11 | Glutaric Acid | Glutaryl-CoA Metabolism |
| 12 | Humic Acid | Soil Organic Matter |
| 13 | Itaconic Acid | Citric Acid Cycle |
Technical Route of Targeted Metabolomics of Organic Acids
Technology Platform for Organic Acid Analysis
- Analytical platform: AB Sciex QTRAP ® 6500 LC-MS/MS
- Detection mode: MRM
- Ion source：ESI
- Detection accuracy: pmol
- Linearity: r>0.99
- Strict quality control
1) Blank: Remove residual impurities in the instrument, provide a stable detection environment, and also monitor the stability of the instrument.
2) Mixed standard: Mixed standard mainly monitors the stability of the instrument.
3) Random sample repetition: monitor the stability of the entire extraction process.
Service Process
Data Analysis Provided by Organic Acid Analysis
| Data Analysis Type | Description |
|---|
| Quantitative Analysis | Accurate quantification of specific organic acids in samples, often reported in units such as mg/L or μg/g. |
| Qualitative Analysis | Identification of the presence or absence of specific organic acids in samples, often used for initial screening. |
| Profiling Analysis | Comprehensive analysis of a sample to identify and quantify multiple organic acids, providing a detailed profile of the sample's composition. |
| Trend Analysis | Monitoring changes in organic acid levels over time, which can be valuable for process optimization or tracking environmental trends. |
| Correlation Analysis | Determining relationships between organic acid levels and other variables, such as environmental factors or product characteristics. |
| Statistical Analysis | Employing statistical methods to analyze organic acid data, including measures of central tendency, variability, and hypothesis testing. |
| Pattern Recognition | Identifying unique patterns or signatures in organic acid profiles that may indicate specific conditions or sources. |
| Multivariate Analysis | Using mathematical and statistical techniques to analyze relationships among multiple variables, including various organic acids. |
| Predictive Modeling | Developing models to predict future organic acid levels based on historical data, which can aid in decision-making and process control. |
| Data Visualization | Presenting organic acid data in graphical formats like charts, plots, and heatmaps to enhance data interpretation. |
| Customized Reporting | Tailoring data reports to meet specific client needs, including summary statistics, graphs, and actionable insights. |
Applications of Organic Acid Analysis
Biotechnology: Monitor organic acid production in microbial cultures for bioproducts.
Metabolomics Research: Gain insights into cellular metabolism for various research fields.
Food and Beverage Industry: Ensure product quality by monitoring acidity levels. Assess fermentation processes in dairy products, wines, and more. Detect organic acids in beverages and food items.
Environmental Monitoring: Evaluate soil and water quality. Identify indicators of pollution or microbial activity.
Chemical Synthesis: Verify reagent purity in chemical reactions.
Agriculture: Study soil health and nutrient content for optimized crop growth.
Pharmaceuticals: Quality control and verification of drug formulations.
Waste Management: Optimize composting and anaerobic digestion processes by assessing organic waste composition.
Petroleum Industry: Detect and quantify corrosive organic acids to prevent pipeline and equipment damage.
Project Cycle
A standard experiment and analysis process takes about 2~6 weeks.
Creative Proteomics provides accurate and detailed data and analytical reports by selecting the appropriate column, such as a C18 column or a HILIC column, depending on the type of organic acid to be analyzed. Please feel free to contact us if you have any questions or specific requests.
Case: Quantitative LC-MS/MS Analysis Reveals Subcellular Distribution of Low Molecular Mass Organic Acids in Osteosarcoma Cells
Background:
The background information emphasizes the importance of low molecular mass organic acids (LMMOAs) in cellular metabolism, particularly their role in the tricarboxylic acid (TCA) cycle and oxidative phosphorylation (OXPHOS). It also highlights the relevance of studying LMMOAs, including malic acid (MA), succinic acid (SA), fumaric acid (FA), and citric acid (CIA), in the context of cancer research and metabolic re-programming.
Samples:
The study primarily focuses on human osteosarcoma 143B-derived wild-type cybrids as the biological samples of interest. These cells were grown in a controlled environment and subsequently subjected to subcellular fractionation, allowing for the isolation of mitochondria and cellular extracts.
Technical Methods:
The study employed a comprehensive set of technical methods:
Standards and Solutions: High-quality reagents, including HPLC-grade acetonitrile, formic acid, and the LMMOAs, were used. Stock solutions of analytes were prepared and stored for further dilution. Working standards for method validation were obtained through dilution.
Cell Growth and Subcellular Fractionation: The biological samples, human osteosarcoma 143B-derived cybrids, were cultured in specific medium conditions. Mitochondria were isolated through a series of centrifugation and homogenization steps.
Extraction of Cellular and Mitochondrial Fractions: Samples underwent cold methanol/acetonitrile extraction to isolate the LMMOAs.
LC-MS/MS Analysis: Chromatographic analysis was performed using a Phenomenex XSelect CSH Phenyl-Hexyl column. LC-MS/MS analysis was conducted with an API 4000-QTrap triple quadrupole interfaced with an electrospray ionization (ESI) source in negative mode. Multiple reaction monitoring (MRM) was used for quantification.
Method Validation: The method underwent comprehensive validation, including linearity studies over specific concentration ranges, determination of limit of detection (LOD) and limit of quantitation (LOQ), recovery experiments, precision assessment, and evaluation of matrix effects.
Results:
The results indicated the successful development and validation of the LC-MS/MS method for the quantification of LMMOAs (MA, SA, FA, and CIA) in cellular and mitochondrial extracts. The method demonstrated excellent linearity, sensitivity, and accuracy. Notably, it enabled the differentiation of LMMOAs' distribution between cellular and mitochondrial fractions. In particular, SA was found to be predominantly abundant in mitochondria, while MA and FA were more prominent in the cytosol. CIA levels were low, likely due to its active utilization in metabolic pathways.
MRM chromatograms
Reference
- Fiori, Jessica, et al. "Cellular and mitochondrial determination of low molecular mass organic acids by LC–MS/MS." Journal of Pharmaceutical and Biomedical Analysis 150 (2018): 33-38.
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