Development of method to assay pepsin activity in gastric juice
 
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Katedra i Zakład Chemii, Wydział Lekarski z Oddziałem Lekarsko-Dentystycznym w Zabrzu Śląskiego Uniwersytetu Medycznego w Katowicach
 
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Laboratorium Analityczno-Bakteriologiczne, Niepubliczny Zakład Opieki Zdrowotnej, Zakład Pulmonologii w Tarnowskich Górach
 
 
Corresponding author
Michał Długaszek   

Katedra i Zakład Chemii Wydział Lekarski z Oddziałem Lekarsko-Dentystycznym w Zabrzu Śląskiego Uniwersytetu Medycznego w Katowicach, ul. Jordana 19, 41-808 Zabrze, tel. +48 693 962 272
 
 
Ann. Acad. Med. Siles. 2015;69:85-90
 
KEYWORDS
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ABSTRACT
Introduction:
The aim of this study was to develop a method to assay the activity of pepsin in gastric juice based on the method described in the references, which was not entirely effective in laboratory tests. The study aims to improve the aforementioned method based on the digestion of bovine hemoglobin by pepsin and to introduce effective changes so that the results may be useful for diagnostic purposes.

Materials and methods:
To achieve this, tests were carried out using the hemoglobin of different species of animals and human hemoglobin, changing the incubation periods, concentrations and volumes of reagents.

Results:
Tests based on the use of purified (crystalline) pepsin revealed that the most effective in the assay of enzyme activity is human hemoglobin. The originally proposed incubation periods were extended, and in order to inhibit the reaction, trichloroacetic acid was used in a higher concentration than suggested, while reducing by half the volume of reactants. In the tests carried out using a purified enzyme, a correlation between the known concentration of the enzyme and the absorbance of supernatant containing the reaction products of enzymatic digestion of hemoglobin was proved.

Conclusions:
Introducting changes leads to more effective diagnostic use.

REFERENCES (12)
1.
Ozmen S., Yücel O.T., Sinici I., Ozmen O.A., Süslü A.E., Oğretmenoğlu O., Onerci M. Nasal pepsin assay and pH monitoring in chronic rhinosinusitis. Laryngoscope 2008; 118: 890–894.
 
2.
Samuels T.L., Johnston N. Pepsin as a marker of extraesophagealreflux. Ann. Otol. Rhinol. Laryngol. 2010; 119: 203–208.
 
3.
Abd El-Fattah A.M., Abdul Maksoud G.A., Ramadan A.S., Abdalla A.F., Abdel Aziz M.M. Pepsin assay: a marker for reflux in pediatric glue ear. Otolaryngol. Head Neck Surg. 2007; 136: 464–470.
 
4.
Anson M.L. The estimation of pepsin, trypsin, papain and cathepsin with hemoglobin. J. Gen. Physiol. 1938; 22: 79–89.
 
5.
Wynn W.A. A Simple Method for the Estimation of Pepsin in Gastric Juice. J. Clin. Pathol. 1955; 8: 85.
 
6.
Balasubramaniam P., Malathi A. Comparative study of hemoglobin estimated by Drabkin's and Sahli's methods. J. Postgrad. Med. 1992; 38: 8–9.
 
7.
Yvon M., Chabanet C., Pellisier J. Solubility of peptides in trichloroacetic acid (TCA) solutions. Hypothesis on the precipitation mechanism. Int. J. Pept. Protein Res. 1989, 34: 166–176.
 
8.
Dembińska-Kieć A., Naskalski J. Diagnostyka laboratoryjna z elementami biochemii klinicznej. Wyd. II uzupełnione i poprawione, Urban & Partner, Wrocław 1998, s. 633–636.
 
9.
Kokot F. Choroby wewnętrzne. Tom I. Wyd. VIII zmienione i unowocześnione. Wydawnictwo Lekarskie PZWL, Warszawa 2006, s. 326.
 
10.
Abu-Erreish G.M., Peanasky R.J. Pepsin inhibitors from Ascaris lumbricoides. Pepsin-inhibitor complex: stoichiometry of formation, dissociation, and stability of the complex. J. Biol. Chem. 1974; 249: 1566–1571.
 
11.
Ofori-Anti A.O., Ariyarathna H., Chen L., Lee H.L., Pramod S.N., Goodman R.E. Establishing objective detection limits for the pepsin digestion assay used in the assessment of genetically modified foods. Regul. Toxicol. Pharmacol. 2008; 52: 94–103.
 
12.
Yamada S., Hirata K., Tsugawa N., Bunai Y., Ohya I. Vomit identification by pepsin assay using a fibrin blue-agarose gel plate. Forensic Sci. Int. 1992; 52: 215–221.
 
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