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BOC Sciences has a professional amino acid analysis laboratory, rich experience in amino acid analysis, and advanced technology to provide customers with fair, rigorous, reliable and efficient amino acid derivatization analysis services.
Amino acids are the basic structural units that make up proteins and peptides, and are also essential nutrients for organisms to maintain growth. They participate in the metabolic process of the body and have a wide range of biological activities and special physiological functions. Based on the importance of amino acids, amino acid analysis has always been an important part of research in the field of analysis. Amino acid analysis plays an important role in research in many fields such as chemistry, bioengineering, food science, and clinical medicine. Amino acid analysis can be divided into two categories according to whether it is derivatized, one is the direct analysis method, and the other is the derivatized indirect analysis method. Most amino acids do not contain chromophores and cannot be directly detected with conventional UV detectors. By derivatizing the amino acid and attaching a chromogenic group, the qualitative and quantitative analysis of the derivatized amino acid derivatives can be carried out, and finally the sensitivity and separation selectivity of the analysis and detection can be improved.
We use pre-column derivatization technology to convert amino acids into derivatives that are suitable for reversed-phase chromatographic separation and can be sensitively detected, and are suitable for amino acid analysis of hydrolyzed peptides and proteins. We can realize RP-HPLC analysis by attaching UV-absorbing groups to amino acids through derivatization reactions. The most commonly used are AQC derivatives and PITC derivatives.
Post-column derivatization is a process in which the sample components are first separated by a chromatographic column, and then derivatized and detected. Derivatization reagents are generally ninhydrin and o-phthalaldehyde. Compared with pre-column derivatization, the advantage of post-column derivatization is the good reproducibility of the reaction. We hydrolyze the sample with hydrochloric acid to form free amino acids, and then use the ion exchange column of the amino acid analyzer to separate them. Amino acids react with ninhydrin solution in color. Subsequently, we determined the amino acid content colorimetrically by a spectrophotometer.
Regardless of the determination of free amino acids or hydrolyzed amino acids, since most amino acids contain at least highly polar amino groups and carboxyl groups, the overall polarity is very high, and the commonly used reversed-phase mobile phases and chromatographic columns are relatively weaker in retention. At the same time, some amino acids lack structural detection features, neither UV absorption nor fluorescence, so the choice of mobile phase and detector is very challenging.
For amino acids, we have replaced active hydrogens on polar functional groups such as hydroxyl, amino, and carboxyl groups with non-polar groups through derivatization, making the analytes more volatile and less reactive, thereby improving their chromatographic behavior. We use a variety of derivatization methods to convert amino acid molecules into substances that absorb ultraviolet and visible light or produce fluorescence, which solves the difficulties in amino acid analysis and improves the sensitivity and resolution of amino acid analysis methods.
