Method for determination of amino acid content in protein products for medical use

Abstract

ABSTRACT. The paper presents a method for determina$ on of amino acid content in collagen biomaterials for medical use produced in the Collagen Department of INCDTP -Division ICPI and its valida$ on. The method has three stages: the fi rst consists in hydrolysing the collagen biomaterial sample down to amino acids; the second step refers to deriva$ za$ on of amino acids with N, O-bis (trimethylsilyl) trifl uoroacetamide (BSTFA) which is a silyla$ on reagent; deriva$ zed amino acids are detected using a mass spectrometer a+ er gas chromatography. The method was validated to establish performance parameters and to check compliance with the intended purpose. KEY WORDS: collagen, amino acids, biomaterials METODĂ PENTRU DETERMINAREA CONŢINUTULUI DE AMINOACIZI DIN PRODUSELE PROTEICE PENTRU UZ MEDICALREZUMAT. Se prezintă o metodă de determinare a conţinutului de aminoacizi din biomaterialele colagenice pentru uz medical produse în Departamentul Colagen al INCDTP -Sucursala ICPI şi validarea ei. Metoda are trei etape: prima constă în hidroliza probei de biomaterial colagenic până la stadiul de aminoacizi; a doua etapă se referă la deriva$ zarea aminoacizilor cu N, O-bis (trime$ lsilil) trifl uoracetamidă (BSTFA) care este un reac$ v de sililare; aminoacizii deriva$ zaţi sunt detectaţi pe un spectrometru de masă după gaz cromatografi e. Metoda a fost validată pentru a se stabili parametrii de performanţă şi pentru verifi carea conformării cu scopul propus. CUVINTE CHEIE: colagen, aminoacizi, biomateriale PROCÉDÉ DE DÉTERMINATION DE LA TENEUR EN ACIDES AMINÉS DANS DES PRODUITS PROTÉIQUES À USAGE MÉDICALRÉSUMÉ. On présente une méthode de détermina$ on de la teneur en acides aminés de biomatériaux de collagène à usage médical fabriqués dans le Département Collagène de INCDTP-ICPI et sa valida$ on. La méthode comporte trois étapes : la première consiste en l'hydrolyse de l'échan$ llon de biomatériau de collagène au stade des acides aminés ; la deuxième étape se rapporte à la dériva$ sa$ on d'acides aminés avec du N, O-bis (triméthylsilyl) trifl uoroacétamide (BSTFA) qui est un réac$ f de silyla$ on ; les acides aminés dérivés sont détectés sur un spectromètre de masse après chromatographie en phase gazeuse. La méthode a été validée pour établir les paramètres de performance et vérifi er la conformité avec l'objec$ f prévu.

Study of the Chemical Composition, Amino Acid, Flavonoids and Vitamin C of Moringaoliefera Leaves Extract Grown in AL-Ramadi- Iraq

“…Sixteen peaks were determined from the chromatogram of the aqueous leaf extract of Moringaoleifera. These peaks indicate the presence of sixteen compounds (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16) in the extract. The molecular formula, percentage content and molecular mass of the compounds are shown in (Table 1).…”

“…Moringaoliefera leaf extract was rich with long chain hydrocarbon (Alkanes, Alkenes) ,alcohols,fatty acids and another compound, For example, the compound Decanoic acid is one of the components that has straightchain saturated fatty acid. It has a role as an antibacterial agent, an anti-inflammatory agent, a human metabolite, a volatile oil component, a plant metabolite and an algal metabolite.Decanoic acid non-toxic ,used to make esters for perfumes and fruit flavors and as an intermediate for food-grade additives (8) . Also the other chemical compound is 17-Octadecynoic acid, it is is an acetylenic fatty acid that is octadecanoi acid (stearic acid) which has been doubly dehydrogenated at positions 17 and 18 to give the corresponding alkynoic acid.…”

“…About 5 mg solid samples were weighed with an accuracy of 0.01 mg and approximately 100 mg liquid samples were weighed with an accuracy of 0.01 mg then 1 ml of 6 M hydrochloric acid solution as hydrolysis agent was added, the tube was covered and placed in the aluminum thermo block at 100°C±20°C for 24 hours for hydrolysis , Using a pipette, a volume of 100 μl of hydrolyzed is introduced in a vial placed in evaporation to remove moisture with nitrogen gas; The dried amino acid residues were dissolved in a volume of 100 μl of acetonitrile; They are derivative with a volume of 100 μl of OPA( orthophethalene aldehyde ); The sealed vial is subjected to ultrasound for 1 minute;• The vial is placed in the thermo block at 100°C+2°C for 30 min. to complete the derivatization reaction; The vial is placed in the gas chromatograph sample stand; 10 injections of 100 μl per sample are performed (8) .…”