Amino Acid Analysis
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Amino Acid Analysis

Amino acid analysis (AAA) involves identifying amino acid types and quantities. It can be utilized to assess the amino acid makeup of proteins, peptides, and protein drug samples, determine protein levels post-hydrolysis, or characterize proteins through peptide mass fingerprinting or in conjunction with MS/MS sequencing. BOC Sciences employs GC/MS and LC/MS platforms to offer comprehensive amino acid analysis services, enabling both quantitative and qualitative analysis of over 70 amino acids and their derivatives. We always put the quality of its scientific research service on top, prioritizes the needs of our customers, performs our analysis under strict procedures, and provides our customers with professional scientific analysis of the research and development services.

Steps in the digestion of proteins into amino acids.Fig. 1. Steps in the digestion of proteins into amino acids.

What Are Amino Acids?

Amino acid is the basic unit of protein. Its wide range of use can be applied to areas such as food, medicine, feed additives, cosmetics, industry and agriculture. Development in bioengineering technologies has helped amino acid to gradually become one of the major research areas in the 21st century. Demand for amino acids is increasing across various industries, and new amino acid reforming processes are emerging. In recent years, as researchers pay attention to the innovation and improvement of amino acid analysis methods, the speed, sensitivity and instrument automation level of amino acid analysis have improved. Currently, the instruments used to measure amino acids mainly include ion exchange chromatography (IEC), high performance liquid chromatography (HPLC) or gas chromatography (GC). Detectors used by these instruments include UV-visible absorption, fluorescence, chemiluminescence, etc.

Amino Acid Analysis Methods

Technologies used to analyze amino acid are important for life science, food science, clinical medicine, geological investigation, paleontology and other fields. Amino acid analysis has a wide range of applications. For example, amino acid analysis can measure the composition and content of amino acids in protein (peptide) drugs, and identify the presence of atypical amino acids in protein (peptide) drugs. This is a necessary step before complete protein sequencing and provides strong evidence for the protein's primary organization. Amino acid composition, mass spectrometry, and amino acid sequence determination are of great significance to confirm the structure of peptides, and amino acid composition analysis is an investigation item that must be added to the quality research of peptide drugs and their raw materials. The following are some techniques commonly used for amino acid analysis:

  • MALDI-TOF MS: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry can be used to analyze and identify amino acids and small peptides.
  • Electrospray ionization mass spectrometry (ESI-MS): Electrospray ionization is often combined with liquid mass spectrometry (LC-MS) to analyze and quantify the amino acid composition of proteins and peptides.
  • Ion exchange chromatography: Separation using the charge characteristics of amino acids.
  • Reversed-phase high-performance liquid chromatography (RP-HPLC): Often used in conjunction with mass spectrometry to achieve accurate quantitative analysis of amino acids.
  • Nuclear magnetic resonance (NMR): Although NMR is typically used to study the three-dimensional structure of proteins, information on the type and environment of amino acids can also be obtained with appropriate one- and two-dimensional NMR experiments.

Amino Acid Analysis Services

BOC Sciences' amino acid analysis solutions provide you with a way to control the main factors that change your amino acid analysis. From optimized instrumentation and quality control chemistry methods to standardized software engineering, methods, report templates, and qualification tools, you can rely on a comprehensive, detailed, robust, and reliable analytical method to ensure successful analysis of every sample.

Our services include but not limited to the following:

Amino acid composition analysis can qualitatively and quantitatively determine the amino acid composition in a sample. We use amino acid automatic analyzer and high-performance liquid chromatography to determine the amino acid composition of various samples. Our one-stop service includes processing and analysis of samples provided by customers, analysis of acquired data and completion of project reports.

We provide N-terminal sequencing services based on mass spectrometry technology and Edman degradation method, which can achieve accurate interpretation of N-terminal amino acid sequences and ensure the coverage and accuracy of sequencing results. The N-terminal sequencing based on the Edman degradation method can realize the determination of 20-40 amino acids at a time. N-terminal sequencing based on mass spectrometry is complementary to Edman degradation sequencing, which can realize the determination of N-terminal blocked amino acid sequence.

We can provide comprehensive C-terminal sequencing services for various samples. The methods of C-terminal sequencing include mass spectrometry, carboxypeptidase method and chemical reagent method. We can complete amino acid C-terminal sequencing within 4-5 weeks using mass spectrometry.

We have a professional amino acid analysis laboratory, which can use a variety of derivatization methods to convert amino acid molecules into substances that absorb ultraviolet and visible light or produce fluorescence. We performed qualitative and quantitative analysis of derivatized amino acid derivatives, solved the difficulties in amino acid analysis, and improved the sensitivity and resolution of amino acid analysis methods.

Without adding ion-pair reagents, we derivatize a large number of amino acids through different separation modes such as reversed-phase chromatography mode, hydrophilic interaction chromatography (HILIC) mode, and ion exchange mode. In addition, we can add volatile ion-pairing reagents to the mobile phase system to perform derivatized direct analysis of amino acids in clinical blood samples.

We employ a variety of analytical methods to provide high-quality solutions for the chiral analysis of protected and unprotected amino acids. We use normal-phase or reverse-phase polysaccharide chiral columns to analyze amino acids protected by groups such as Boc or FMOC, and use ligand-exchange chiral columns to analyze unprotected amino acids. Appropriate mass spectrometry methods as well as capillary electrophoresis based on chiral ligand exchange also enable chiral separation and analysis of amino acids.

We can provide high-resolution, high-sensitivity, high-speed, high-throughput UPLC amino acid analysis solutions, and issue UPLC analysis and detection data in a short time. Our UPLC amino acid analysis solution is suitable for the characterization of amino acids in peptides and proteins and the analysis of amino acids in food, feed, pharmaceuticals and cosmetics.

With years of experience in mass spectrometry services, we use a variety of mass spectrometry techniques to identify the type and molecular weight of amino acids, and identify the composition and arrangement information of amino acids. We can detect regular free amino acids as well as amino acids of physiological importance. In addition, we can also customize the detection of special amino acids according to the specific needs of customers.

Based on tandem mass spectrometry, de novo sequencing is a valuable method for obtaining peptide and protein amino acid sequences without prior knowledge.

What Can We Offer?

BOC Sciences possesses extensive technical expertise and solutions in amino acid quality research and amino acid composition drugs. This includes setting standards for starting materials of active pharmaceutical ingredients, conducting quality research and filing for active pharmaceutical ingredients, developing amino acid preparation prescriptions, and conducting quality research on peptide/protein drugs (such as amino acid composition, extinction coefficient, sequence determination, D-configuration determination, peptide mapping analysis, etc.).

  • Quality Research on Amino Acid Raw Materials and Preparations

  • Structural analysis of unidentified impurity peaks (2D-LC/QTOF and Pre-HPLC/NMR).
  • Development of methods for analyzing impurity spectra in raw materials and related substances (reverse-phase or HILIC method).
  • Achiral/chiral isomer impurity method development services (derivatized/non-derivatized methods).
  • Trace analysis of raw materials and preparations, including research on solvent residues, genotoxic impurities, and elemental impurity risks.
  • Investigation of enzyme catalysis and fermentation residues: protein/enzyme residues, host DNA residues, resin leachates, and defoaming agent residues.
  • Stability research and API filing and registration services.
  • Development of compound amino acid formulations and assistance with product registration.
  • Quality Research on Amino Acid of Peptide or Protein Products

  • Determination of extinction coefficient.
  • Quantitative analysis services: concentration determination.
  • Amino acid and protein isoelectric point determination.
  • D-isomer quantification service: deuterated acid hydrolysis-pre-column derivatization LCMS method.
  • Protein characterization: amino acid sequencing, N/C-terminal sequencing, MS peptide map analysis, disulfide bond analysis, glycoform analysis, etc.
  • Analysis of total amino acid composition, including cysteine, tryptophan, and hydroxyproline.
  • Free amino acid analysis, including asparagine and glutamine.
  • Structural identification of unknown structural impurity peaks.

Why Choose BOC Sciences?

  • Detection of all 20 natural and unnatural amino acids with high sensitivity down to the microgram level.
  • More than 15 years of experience in new drug discovery, analysis and compliance laboratory quality management.
  • Having extensive experience in the analysis of biopharmaceuticals, including amino acids, peptides, and proteins.
  • Having international standard instruments and equipment such as high-resolution mass spectrometer, triple quadrupole, two-dimensional liquid phase mass spectrometry, quadrupole mass spectrometry, MALDI-TOF, GC-MS, and ICP-MS.
  • Stable and reliable quality, competitive cost, and comprehensive report provided within 5-7 days.

Amino Acid Analysis by HPLC

Amino acid analysis by HPLC


  1. Piestansky J, Galba J, Olesova D, Kovacech B, Kovac A. Determination of immunogenic proteins in biopharmaceuticals by UHPLC-MS amino acid analysis. BMC Chem. 2019;13(1):64. Published 2019 May 10. doi:10.1186/s13065-019-0581-z.
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