Тebl 4.
Qualitative and quantitative analysis of amino acids in collagen (100g)
Amino acid
|
In literature [17. P. 40]
|
In the study
|
Difference
|
Histidine
|
2,60
|
2,63
|
+0,03
|
Asparagine
|
0,42
|
0,44
|
+0,02
|
Aspartate
|
4,45
|
4,52
|
+0,07
|
Threonine
|
4,90
|
4,90
|
-
|
Serine
|
1,87
|
1,87
|
-
|
Glutamine
|
3,87
|
3,88
|
+0,01
|
Proline
|
7,19
|
7,23
|
+0,04
|
Glycine
|
11,82
|
11,97
|
+0,15
|
Valine
|
33,50
|
35,12
|
+1,62
|
Methionine
|
10,93
|
11,00
|
+0,07
|
Isoleucine
|
2,02
|
1,87
|
-0,15
|
Leucine
|
0,61
|
0,56
|
-0,05
|
Tyrosine
|
1,36
|
1,36
|
-
|
Phenylalanine
|
2,66
|
2,74
|
+0,08
|
Hydroxyproline
|
0,52
|
0,51
|
-0,01
|
Hydroxylysine
|
1,31
|
1,34
|
+0.03
|
Valine
|
9,21
|
10,69
|
+1,48
|
Methionine
|
0,76
|
0,74
|
-0,02
|
From the data presented in Table 4, it can be seen that the amino acid composition and the amount of both collagen proteins are almost the same. There are partial differences in some individual amino acids, which are mainly related to the separation process. This does not greatly affect its properties. It was also found that the most common amino acids in the composition of collagen practically did not differ in the amounts of glycine, proline and hydroxyproline.
Conclusion. The most optimal conditions for the isolation of the aqueous mass of collagen from the skin of cattle were found by the method of alkaline-salt hydrolysis without additives, the natural structure of which is preserved. After buffer extraction, a second hydrolysis was performed with 5% NaOH and 80 g/L Na2SO4 for 80 min. The results are validated by specific gravity, solubility and viscosity methods. It was also found that the most common amino acids in the composition of collagen practically did not differ in the amounts of glycine, proline and hydroxyproline..
Experimental part. Extraction of the aqueous mass of collagen. The first known hydrolysis method was used to obtain an aqueous mass of collagen by alkaline-salt hydrolysis of cattle skin [11. P. 14]. The second time the hydrolysis was carried out as follows. First washed in distilled water until neutralized, sorted; Hydrolysis in a 5% sodium hydroxide solution in a ratio of 1:10 All processes were carried out at a temperature of +21 - +230C.
Determination of the viscosity of the water mass of collagen. To determine the viscosity of collagen in 6% acetic acid 0.01; 0.02; 0.025; 0.03; 0.0375; Prepared 0.05% solutions. The dependence of the viscosity of collagen solutions on the concentration of solutions was determined on a Ubellode viscometer [ƞ]. The definition of intrinsic viscosity is calculated using the following formula:
Here:
tn – is the solution flow time
t0 – is the solvent flow time
Cn – is the concentration of the solution
Amino acid analysis. The amino acid composition of collagen was determined by the Cohen method using HPLC, in the form of PTC derivatives of amino acids [18. S. 1-16].
References
1. Мильчевский Ю.В, Жоров Б.С, Есипова Н.Г, Туманян В.Г.Структура коллагена с новой сеткой водородных связей // Биоорганическая химия. –1999. –Т. 25. –№5. –С. 348-357.
2. Rémi Parenteau-Bareil, Robert Gauvin and François Berthod. Collagen-Based biomaterials for tissue engineering applications // Materials. –2010. –V. 3. –P. 1863-1887.
3. E. J Miller., K. A Piez and A. H Reddi. Biomedical and industrial application of collagen; In Extracellular Matrix Biochemistry eds // Elsevier, New York. –1984. –V. 24.–P. 41-81.
4. Drake M.P., Davison P.F., Bumps S., Schmitt F.O. Action of Proteolytic Enzymes on Tropocollagen and Insoluble collagene // Biochemistry. – 1996. – Vol. 5. – № 1. – P. 301–312.
5. Charriere G., Bejot M., Schnitzler L., Ville G., Hartmann D.J. Reactions to a bovine collagen implant Clinical and immunologic study in 705 patients // J. Am. Acad. Dermatol. –1989. –V. 21. –P. 1203–1208.
6. Badylak S.F., Gilbert T.W. Immune response to biologic scaffold materials // Semin. Immunol. –2008. –V. 20. –P. 109–116.
7. Антипова Л. В., Сторублевцев С. А. Сравнительные свойства коллагеновых белков рыбного и животного происхождения // Вестник ВГУ, серия: химия. биология. Фармация. –2016. –№ 4. –С. 47-54.
8. Королева О.В., Машенцева Н.Г., Митасева Л.Ф., Семенышева А.И., Николаев И.В., Степанова Е.В. Биотрансформированное мясное сырье в качестве белкового композита в производстве паштетов // Fleischwirtschaft-International Россия. –2012. –№ 2. –C. 63-66.
9. Иванкин А.Н., Панферов В.И., Фахретдинов Х.А., Вострикова Н.Л., Куликовский А.В., Голованова П.М//Наномикрокомпозиционные раневые покрытия на основе коллагена и карбоксиметилцеллюлозы. Лесной вестник / Forestry bulletin. –2015. –№1. –С. 41-45.
10. Сапожникова А.И., Каспарьянц С.А., Белевцова Д.В//Способ получения коллагенового золя. Пат. РФ №2129805.
11. Истранов Л.П., Абоянц Р.К., Белозерская Г.Г., Макаров В.А//Местные гемостатические средства на основе коллагена//Технология лекарственный средст. –2007. –№7. –C.29-32.
12. Гулямов. Т., Раджабов О.И., Тураев А.С., Атаджанов А.Ю // Способ получения субстанции коллагена. Патент Uz IAP 04736. –2003.
13. Гулямов Т., Раджабов О.И., Атажанов А.Ю., Муйдинов Н.Т., Халилова Г.А., Тураев А.С // Способ выделения коллагена и создание препаратов на его основе пленок. ДАН РУз. 2017. №6. С.52-56.
14. Гулямов Т., Муйдинов Н.Т., Атажанов А.Ю., Жумаева Ш.Х., Шомуротов Ш.А., Раджабов О.И., Тураев А.С// Способ получения апликационного средства. Патент РУз № IАР 05873. –2019.
15. Radjabov O.I, Muydinov N.T,. Gulyamov T, Turayev A.S. Otajonov A.Y. Native collagen and its hemostatic form // Программа и тезисы научно-практической конференции молодых ученых посвященной 110-летию академика С.Ю.Юнусова Актуалные проблемы химии природных соединений. Тошкент. 2019. с. 111-С.
16. V. Yannas, J. F. Burke, P. L. Gordon, C. Huang, and R. H. Rubenstein//Design of an Artificial Skin. II. Control of Chemical Composition. Journal of Biomedical Materials Research. Vol.14. P. 107-131 (1980).
17. Антипова Л. В., Сторублевцев С. А. Сравнительные свойства коллагеновых белков рыбного и животного происхождения // Вестник ВГУ, серия: химия. биология. Фармация. –2016. –№ 4. –С. 47-54.
18. Steven A., Daniel J., Strydom D. Amino acid analysis utilizing fhenylisothiocyanate derivatives // Analytical Biochemistry.–1988. –No.1. (17). –P. 1–16.
AUTHOR BIOGRAPHIES
Ibragim R. Askarov, Andijan State University
Doctor of Chemical Sciences, Professor of the Department of Chemistry, Honored Inventor of the Republic of Uzbekistan, Chairman of the "TABOBAT" Academy of Uzbekistan
Nurillo T. Muydinov, Andijan State University
PhD, Senior Lecturer of the Department of Chemistry
Do'stlaringiz bilan baham: |