GRINSPANA-FONNJESUE METHOD (E. M. Greenspan, V. Fonnesu) — quantitative definition of mukoproteid of blood serum. Modification of Fonnezyu (1960) original methods of Greenspan with coauthors (1952) consists in change of the standard structure of a biuretic reactant. With the diagnostic purpose the maintenance of mukoproteid in blood serum is defined at rheumatism, malignant new growths, a thyrotoxicosis.
The method is based on dissolution of mukoproteid in chloric to - those (other proteins in these conditions drop out in a deposit) with the subsequent sedimentation their phosphatotungstic to - that. The amount of protein in the deposit dissolved in the diluted alkali is defined colorimetric with a biuretic reactant.
The course of definition
For extraction of mucoproteins of 2,0 ml of blood serum mix from 4,0 ml 6% of solution chloric to - you and filter via the wet filter within 1 hour (not less). Add 0,8 ml of 5% of solution to the received filtrate phosphatotungstic to - you (H 3 PO 4 - 12WO 3 • 12H 2 O), prepared on 2 N solution salt to - you. In 10 min. centrifuge (10 min., 2000 about I mines). The deposit is dissolved in 1 ml 0,4 N of solution of caustic soda, add 0,4 ml of a biuretic reactant (10 ml of 1% of solution of a copper vitriol — CuSO 4 • 5H 2 O — mix from 50 ml 15% of NaOH solution). Coloring develops within 1 hour in the dark. The optical density of the painted solution is measured at 630 nanometers. Control is prepared as follows: add 0,4 ml of a biuretic reactant to 1 ml 0,4 N of NaOH. Standard solution (200 mg of casein dissolve in 100 ml 0,4 N of NaOH solution) is processed as pilot test. Calculation is made on a formula:
[E of pilot test • 100] / [E of standard test] = quantity of mukoproteid, in mg of %,
where E — the size of an extinction found at a kolorimetrirovaniye. (At the healthy person in blood serum from 40 to 75 mg of % of mukoproteid are defined by this method.)
The quantity of mukoproteid in blood serum increases at rheumatic process (in proportion to increase in its activity), and also at malignant new growths and hyperfunction of a thyroid gland.
The described method differs in specificity, however its practical value decreases because of need of use of a large amount of blood serum (2 ml) and labor inputs of statement.
See also Blood, methods of a research .
Bibliography: Biochemical methods of a research in clinic, under the editorship of A. A. Pokrovsky, page 244, M., 1969; Kuzmakn. I. Comparative assessment of some methods of definition of seromucoid in blood serum, Laborat, business, JVe 3, page 146, 1970, bibliogr.; Methodical instructions on use of the unified clinical methods of a research, under the editorship of V. V. Menshikov, M., 1973; Fonnesu V. La determinazione delle mucoproteine (sieromucoide) del siero nell’ ambito delle epatopatie, Minerva med., y. 51, p. 4234, 1960, bibliogr.; G r e e n s p a n E. M. a. o. Serum mucoproteins as aid in differentiation of neoplastic from primary parenchymatous liver disease, J. Lab. clin, med., v. 39, p. 44, 1952.
L. H. Delektorskaya.