Content Determination of Amino Acids Using Ion Chromatography

There are currently 22 amino acids determined as protein building blocks. Moreover, researchers have discovered roughly 250 non-proteinogenic naturally occurring amino acids. Meteorites and comets have also been shown to have amino acids. The standard 20 amino acids, which are stored in the form of codons in the DNA.

Out of these standard amino acids, the human body or the microorganisms existing in the human digestive system synthesize 12 amino acids and the remaining eight essential amino acids are obtained from food. Conversely, all the amino acids required for plants and microorganisms are synthesized by themselves.

The human health is largely affected by the concentrations of various amino acids. Therefore, it is essential to perform amino acid analysis in different fields. For instance, in the clinical field, the amino acid analysis in physiological fluids like urine or saliva helps diagnose metabolic disorders.

Measuring amino acids in protein hydrolysates helps achieve control over feedstuffs and foods (Figure 1). In the beverage industry, effective process control and media production for cell cultures and fermentation can be achieved through determination of amino acid concentration. Amino acids also play crucial role in producing cheese with the required taste.

Typical application fields for the determination of amino acid contents: food, feedstuff, and health care industry.

Figure 1. Typical application fields for the determination of amino acid contents: food, feedstuff, and health care industry.

Amino acid analysis

Metrohm offers Professional Ion Chromatography (IC) instruments for the analysis of amino acids, anions and cations. VIS detection subsequent to derivatization with ninhydrin is involved in the standardized determination method, which complies with USP <1052> (Method 1).

The Amino Acid Analyzer (Figure 2) is equipped with various modules and an ion chromatograph with gradient capability as the key component. The system uses the new Metrosep Amino Acids 1 - 100/4.0 column to separate amino acids by a binary gradient.

Metrohm

Figure 2. Metrohm's fully automated Amino Acid Analyzer consists of 850 Professional IC, 886 Professional Thermostat / Reactor, 887 Professional UV/VIS Detector, 872 Extension Module IC Pump, and sample changer 889 IC Sample Center.

The 886 Professional Thermostat / Reactor ensures the required temperature of 120°C for post-column derivatization. An additional pump is provided to supply the reagent for postcolumn derivatization. The 887 Professional UV/VIS Detector determines the colored reaction products.

The sample changer used for handling liquid samples composed of amino acids is the 889 IC Sample Center with cooling option. Many of these amino acids do not have temperature resistance. Up to 384 samples can be handled using differently sized sample vessels and microtiter plates in accordance with the SBS standard.

With these capabilities, Metrohm’s Amino Acid Analyzer can perform a fully automated amino acid analysis. It is also possible to analyze biologically active samples because the system features metal-free components for handling samples and eluent.

As a result, artifacts caused by the interactions between metal ions and amino acids and peptides can be prevented. Besides performing routine measurements, the reliable analyzer is flexible enough to perform method development. Figure 3 illustrates a chromatogram of the amino acid content determination.

About Metrohm

At Metrohm is one of the world’s most trusted manufacturers of high-precision instruments for chemical analysis. Metrohm was founded in 1943 by engineer Bertold Suhner in Herisau, Switzerland. Today, Metrohm is represented in 120 countries by subsidiaries and exclusive distributors. The global Metrohm Group also includes the Dutch companies Metrohm Applikon and Metrohm Autolab, manufacturers of online analyzers and instruments for electrochemical research, respectively. Recently, the Metrohm Group was joined by Metrohm Raman, a leading manufacturer of handheld Raman spectrometers.

Metrohm is the global market leader in analytical instruments for titration. Instruments for ion chromatography, voltammetry, conductivity, and stability measurement make the Metrohm portfolio for ion analysis complete. Instruments for Near-infrared and Raman spectroscopy are another, strongly growing segment of the Metrohm portfolio.

Metrohm is a problem solver, both in the laboratory and within the industrial process. To this end, the company offers their customers complete solutions, including dedicated analytical instrumentation as well as comprehensive application know-how. More than 30% of the company’s employees at the Metrohm international headquarters in Herisau work in R&D.

Metrohm has been owned 100% by the non-profit Metrohm Foundation since 1982. The Metrohm Foundation, which does not exert any influence on the company’s business operations, sponsors gifted students in the natural sciences, supports charitable and philanthropic purposes and, above all, ensures the independence of the company.


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Last updated: Apr 16, 2019 at 10:16 AM

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