Catecholamine Analysis Service

v-innovate Technologies can provide isotope coded derivatization-dispersion liquid-liquid microextraction/ultra-high performance liquid chromatography-tandem mass spectrometry and liquid chromatograph mass spectrometer (LC-MS) method for highly sensitive detection of catecholamines, which can obtain comprehensive and high-quality analysis of catecholamine compounds and their metabolites. The identification and precise quantitative analysis of catecholamine compounds and their metabolites have important scientific significance for basic research in neuroscience, brain science, pharmacology, etc.

Overview

Catecholamines are substances containing catechol and amine groups, which play an important role in brain and nerve signal transmission, and are closely related to human health and Parkinson's disease and other neurological and mental diseases. Catecholamines mainly include epinephrine, norepinephrine, isoproterenol, epinephrine, dopamine and related derivatives. These substances are not only important neurotransmitters in the body, but also important hormones, which are closely related to a variety of physiological and pathological phenomena. Related, it plays a key role in maintaining the body's homeostasis balance. The content of catecholamines in biological samples is low, the matrix is complex, and the response signal of its own fluorescence or mass spectrometry detection is not strong, and the sample after isotope derivatization can have high sensitivity, good selectivity, strong anti-matrix interference ability, and high-throughput and accurate determination in LC-MS/MS with the characteristics of simultaneous detection of multiple components. We use d₀/d₃-10-methyl-acridone-2-sulfonyl chloride (d₀/d₃-MASC) as the isotope-encoded derivatization reagent, combined with ultrasonic-assisted dispersion liquid-liquid microextraction technology and UPLC-MS/MS rapid analytical method for the determination of catecholamines. We can also use LC-MS to obtain the content analysis of citrulline, and it provides high sensitivity for its detection.

Figure 1. Overview of the catecholamine autotoxicity theory. (Goldstein 2018).

Applications of Catecholamine Analysis

• Quantitative analysis and application of catecholamine acid and its metabolites
• Promote basic theoretical research in neuroscience and brain science
• Speed up research and application of catecholamine pharmacology and nutrition
• Increase understanding of the mechanism of action of catecholamines and their metabolites
• Enhance pathological research involving catecholamines in biochemical research

Advantages of Our Catecholamine Analysis Service

• Analytical technology is strong against matrix interference
• Use triple quadrupole mass spectrometer to obtain high-sensitivity analysis report
• High sensitivity and high precision analysis results
• Simplified operation process and fast turnaround time
• UPLC-MS/MS platform for accurate determination of catecholamines and their metabolites

Service Workflow

The d0-MASC and d3-MASC were used to derive the microanalysis fluid sample and the reference substance, and the mixture was enriched and purified by the dispersion liquid-liquid microextraction technique, and then detected by UPLC-MS/MS, and quantified with the internal standard substance of the derivative. Samples can be quickly separated and detected, with good peak shape and resolution. LC-MS is used for detection, quantitative analysis results of trace sample substances.

Figure 2. Catecholamine analysis service workflow.

Detection method: standard curve method, isotope coding derivation

Mobile phase: A is methanol, B is 0.1% (volume fraction) formic acid water

Gradient elution conditions: 0-2 min: 50%-90%A; 2-3 min: 90%-100%A; 3-5 min: 100%-50% A

Injection volume: 5 μl

Flow rate: 0.5 ml/min

Elution mode: gradient elution

Analysis time: 3 min

Detection limit: 0. 005-0.010 nmol/l

Correlation coefficient: ≥ 0.995

Lower limit of quantification: 0.018-0.040 nmol/l

Reactive ion: positive ion electrospray ionization (ESI)

Monitoring mode: Multi-reaction monitoring mode of triple quadrupole mass spectrometry

Capillary voltage: 3.0 kV

Desolventizing gas (nitrogen) flow rate: 800 l/h

Cone gas (nitrogen) flow rate: 50 l/h

Collision gas (helium) flow rate: 0.16 ml/min

Retention time: ≤ 0.6 min

Standard deviation: 3.7%

Average matrix effect: 99.3%

Analysis content:

• Construction of standard straight lines
• Isotope code derivation and sample pretreatment
• Ultrasound-assisted dispersion liquid-liquid microextraction
• Optimization of UPLC-MS/MS conditions and quantitative methods
• Optimization of mass spectrometry acquisition conditions for derivatives
• Optimization of UA-DLLME conditions
• Precision, accuracy and stability test
• Linearity, detection limit and lower limit of quantification analysis
• Matrix effect analysis
• Chromatogram raw image and data collection
• Determination of catecholamine and its metabolites

List of Detectable Catecholamines at v-innovate Technologies

Sample Preparation

• Animal and clinical tissue samples: 50mg/sample
• Blood samples (serum, plasma and whole blood): 50ul/sample
• Urine sample: 50ul/sample
• Stool and intestinal contents: 25mg/sample
• Body fluid samples (cerebrospinal fluid, saliva, etc.): 25mg/sample
• Plant tissue samples (roots, stems, leaves, fruits, etc.): 50mg/sample
• Cells and microbial cells: 1x10⁶/sample
• Medium and fermentation broth: 50ul/sample
• Sample storage and transportation: Store in liquid nitrogen or -80°C, and transport on dry ice.

Clinical samples, no less than 30 duplicates in a single group; animal samples, no less than 9 duplicates in a single group; cell and microbial samples, no less than 5 duplicates in a single group; other such as plant samples, no less than 7 duplicates in a single group repeat.

The samples include multiple sample types such as animals, plants and microorganisms, please contact us for details.

Delivery

• Comprehensive experimental procedure
• Sample pretreatment and derivatization treatment
• Optimization of chromatographic conditions and optimization of mass spectrometry acquisition conditions of derivatives
• The most selective analysis of HPLC-MS equipment parameters
• Chromatogram raw image and data analysis report
• Determination of catecholamines and their metabolites
• Customized analysis report

We use isotope coding derivatization-dispersive liquid-liquid microextraction/UPLC-MS/MS and LC-MS method to realize the analytical application of catecholamines. This method has good sensitivity and anti-matrix interference ability, and the method is accurate, fast, specific, and effective for detection. The content of catecholamines in multiple sample types can provide a good technical means for related medical tests and drug screening. v-innovate Technologies is committed to providing you with more competitive analysis costs and rapid analysis technical methods to meet your high-throughput and high-sensitivity detection needs.

References

1. Goldstein D S, Kopin I J. Linking Stress, Catecholamine Autotoxicity, and Allostatic Load with Neurodegenerative Diseases: A Focused Review in Memory of Richard Kvetnansky. Cellular and Molecular Neurobiology. 2018;38(1):13-24.
2. Fernstrom J D, Fernstrom M H. Tyrosine, phenylalanine, and catecholamine synthesis and function in the brain. The American Journal of Psychiatry. 2007;137(6 Suppl 1):1539S-1548S.