Order number: P2145.0005
Delivery time 14 Workdays
The amino acid analysis by oxidative hydrolysis is used when cysteine and methionine in proteins or peptides are to be quantified, since both amino acids are subject to decomposition due to side reactions in hydrochloric acid. In simple total hydrolysis in 6N HCl, methionine oxidizes with partial oxidation to methionine sulfon, which is not stable in hydrochloric acid solution. For that both amino acids must be converted into acid-stable derivatives. At Genaxxon this is done by using performic acid for quantitativ oxidation of Cystein and Methionine. The performic acid oxidizes cysteine completely to cysteic acid and methionine is completely converted into methionine sulfone. Both compounds are stable in hydrochloric acid solution.
After the oxidation step with peramic acid, the analysis sample can then be subjected to total hydrolysis with 6N HCl. After hydrolysis is completed, the hydrolyzate is dried, taken up in buffer and the buffered solution injected onto a HPLC column for separation of the individual amino acids. At Genaxxon, the amino acids are detected by post-column derivatization with ninhydrin. The result is made available to our customers in a detailed report sent as a pdf and excel file.
The amino acid analysis service encloses the total hydrolyis of a given sample, the separation of the amino acids by reversed phase HPLC, the chemical derivatisation of the free amino acids with Ninhydrin (either produced by the acid hydrolysis or of already free amino acids), and transfer of a comprehensive analysis report as pdf-file.
Pure samples are required. The presence of salts, buffers, or detergents are deleterious. Amines (primary or secondary) will react with the carbamate, adversely affecting results. While salts, especially strong buffer substances can alter the pH of the sample causing the hydrolysis/derivitisation to be incomplete or simply fail. Additionally, significant levels of glycerol or carbohydrates are problematic - the glycerol is nonvolatile and attracts moisture (acid) and carbohydrates char, decomposing to ash taking the sample with them. Samples containing Chitin or samples from bones, cartilage or blood plasma can be normally analysed without problems.
For analysis of proteins we do need 25µg to 100µg (500pmol to 2nmol) based on the molecular weight of each protein. For peptides, the requirement is much smaller - 5µg to 20µg (5nmol to 25nmol) will be ok.
The total hydrolysis (P2145.0001 >) is necessary when the analysis of bound amino acids (Proteins or peptides) is conducted. As Methionin and Cysteine are destroyed during total hydrolysis, we offer the oxidative total hydrolysis for quantification of these amino acids (P2145.0005 >). No total hydrolysis is required for the analysis of free amino acids (P2145.0002 >).
Sicherheits Hinweise / Safety
Klassifizierungen / Classificationeclass-Nr: 25-14-10-90
Dokumente - Protokolle - Downloads
Here you will find information and further literature on Amino Acid Analysis of given Sample (oxidative hydrolysis). For further documents (certificates with additional lot numbers, safety data sheets in other languages, further product information) please contact Genaxxon biosience at: email@example.com or phone: +49 731 3608 123.
Hier finden Sie Artikel und Literaturzitate, in denen die Autoren auf die hohe Qualität dieses Genaxxonprodukts vertrauen.
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