Amino Acid Analysis Service

What is Amino Acid Analysis

Amino acid analysis is a fundamental technique in biochemical research and drug development, providing crucial insights into the composition, structure, and function of proteins and peptides. By accurately determining the amino acid sequence and content, this method plays a pivotal role in understanding protein function, identifying potential therapeutic targets, and ensuring the quality and purity of pharmaceutical products.

Creative Proteomics is a reliable partner of biopharmaceuticals. We provide advanced instruments and analytical workflow, aiming to provide you with reliable, high-quality analysis and perfect pre-sales and after-sales services. Amino acid analysis is the key in ICH Q6B specification. We will provide you with amino acid analysis services according to PharmEu and USP <1047>.

We Can Provide but Not Limited to

Amino acid sequence analysis
Amino acid composition analysis
Terminal amino acid sequence analysis
Determination of total amino acids
Determination of free amino acids
Amino acid content analysis
Determination of extinction coefficient
Identify atypical amino acids present in protein/peptide drugs
Qualification of synthetic peptides
Decision support for choice of proteases for protein fragmentation

Technology Platform of Amino Acid Analysis Service

Creative Proteomics provides efficient and accurate protein (peptide) drug amino acid composition analysis services through amino acid derivation technology and high-performance liquid chromatography (HPLC) or thin layer chromatography (TLC) analysis.

We provide the following analysis methods:

1) Hydrolyze protein and peptide samples into single amino acids. Then perform pre-column derivation. The resulting amino acid was derivatized with phenyl isothiocyanate (PITC). The obtained derivative products were subjected to gradient elution with HPLC or TLC to separate different amino acid derivatives. The content of each amino acid derivative is measured by the external standard method, and then the composition and content information of the amino acid in the protein sample is obtained.

2) High performance cation exchange separation is used, followed by post-column Ninhydrinor phthalaldehyde derivatization for amino acid detection. Post-column detection techniques can be used for samples that contain a small amount of buffer components, and each analysis usually requires 5 to 10 ug of protein sample.

Advantages of Amino Acid Analysis Service

Comprehensive services: We provide detailed amino acid profiling across a wide range of biological samples, including serum, plasma, and tissue.
High sensitivity: Low protein/peptide content can be detected to microgram level.
Optimized analysis platform: The current technology uses automatic chromatographic instruments designed for analysis, which can reduce the scale of analysis and greatly improve the flexibility and effectiveness of analysis.
Advanced equipment: High-performance liquid chromatography LC-20AT, SHIMADZU.
Customized service: We can customize exclusive solutions for you based on your samples. You can select or propose the required items for analysis.
Fast Turnaround: We prioritize efficiency without compromising quality, delivering results promptly to meet your project timelines.

Creative Proteomics' analytical scientists can provide our customers with complete confirmation of amino acids in protein/peptide samples, as well as quantitative information on relevant amino acids. Provide quick turnover, clear and concise written reports and customized services to help customers solve analytical and technical problems.

Sample Requirements for Amino Acid Analysis

Sample Type	Amount Required	Storage Conditions	Preparation Notes
Purified Proteins	5-10 µg	Store lyophilized at -20°C	Ensure buffer components do not interfere with analysis
Peptides	5-10 µg	Store lyophilized at -20°C	Desalt if necessary, avoid detergents
Synthetic Peptides	1-5 mg	Store lyophilized at -20°C	Ensure purity and avoid interfering reagents
Biological Fluids	100-200 µL	-80°C	Centrifuge or filter to remove debris
Tissue Lysates	Varies (protein content required)	-80°C	Ensure homogenization, free of contaminants

FAQ

Amino acid analysis services have garnered significant attention as an essential tool in the characterization of proteins and peptides across diverse scientific fields. However, due to the complexity of protein structures and the precision required in the analysis, clients have raised many concerns. To address these common inquiries, we have compiled a comprehensive FAQ section to provide clear and detailed responses, helping to ensure that your experience with our amino acid analysis services is both seamless and informative.

Q: What types of samples can be analyzed in amino acid analysis?

A: We can analyze a wide range of sample types, including purified proteins, peptides, biological fluids, and synthetic peptides. If your sample is complex or contains interfering substances, we can assist with sample preparation or recommend suitable methods for optimal results.

Q: What are the detection limits of your amino acid analysis service, and can it handle low concentrations of proteins or peptides?

A: Our amino acid analysis service offers high sensitivity, capable of detecting protein/peptide content down to the microgram level. This allows for accurate analysis even with low-concentration samples, ensuring precise quantification and composition determination.

Q: Can I customize my amino acid analysis based on specific research needs?

A: We offer tailored analysis services to meet your specific requirements. Whether you need to analyze atypical amino acids, determine free or bound amino acids, or require assistance with synthetic peptide validation, we can design a custom solution for you.

Case Study

Case: Fast and Sensitive Quantification of AccQ-Tag Derivatized Amino Acids and Biogenic Amines by UHPLC-UV Analysis from Complex Biological Samples

Background

The study presents a novel method combining Ultra-High-Performance Liquid Chromatography (UHPLC) with mass spectrometry for the simultaneous analysis of amino acids and biogenic amines. This approach addresses the need for rapid, sensitive, and accurate quantification of these compounds in body fluids, specifically serum and tears. The method aims to enhance the understanding of biochemical profiles and potential biomarkers in various physiological and pathological conditions.

Methods and Instrument

Preparation of Standard Solutions and QC Samples: Calibration standards were created through serial dilutions from this stock. Quality Control (QC) samples were prepared by spiking different matrices (MilliQ water, serum, and tears) with specific concentrations of analytes.

UHPLC-MS Analysis: Liquid chromatography was performed on an Acquity H-class UPLC system with a gradient elution profile. Detection was done using a PDA detector set to 260 nm, and mass spectrometry was conducted on a 5500QTRAP mass spectrometer with electrospray ionization.

Method Validation: The method was validated for linearity, accuracy, precision, recovery, matrix effects, limit of detection (LOD), limit of quantification (LOQ), and stability following FDA guidelines.

Results

The method successfully quantified 35 derivatized amino acids and biogenic amines with good accuracy and precision, meeting FDA standards.

It was effectively applied to analyze serum and tear samples, showcasing its utility in profiling these compounds in body fluids.

The method's robustness and efficiency make it a valuable tool for analyzing changes in amino acid and biogenic amine profiles in various biological contexts.

Chromatogram showing separation of 33 derivatized amino acids and biogenic amines.

Figure 1. Representative chromatogram of the 33 derivatized amino acids and biogenic amines.

Bar graph displaying concentrations of amino acids and biogenic amines in serum.

Figure 2. Concentration of the quantified amino acids and biogenic amines in serum in µmol/L.

References

Guba, Andrea, et al. "Fast and sensitive quantification of AccQ-tag derivatized amino acids and biogenic amines by UHPLC-UV analysis from complex biological samples." Metabolites 12.3 (2022): 272.
Chang J Y, Knecht R. Amino Acid Analysis Using The Dimethylaminoazobenzene Sulfonyl Chloride (Dabs-C1) Precolumn Derivatization Method. Laboratory Methodology in Biochemistry: Amino Acid Analysis and Protein Sequencing, 2019.
Wahl O, Holzgrabe U. Amino acid analysis for pharmacopoeial purposes. Talanta, 2016, 154: 150-163.

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