RU2761045C1 - Method for determination of iron content in muscle tissue of fish - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to animal husbandry, ecology and veterinary medicine. Microelement analysis of fish scales is carried out. The concentration of one or more elements in the flake is detected, optionally Fe and/or Mg. Then the regression equations are calculated according to the original formulas to calculate the concentration of iron in the muscles.

EFFECT: method is lifelong, non-invasive, practical and affordable to use.

1 cl, 3 tbl, 1 ex

Description

This invention relates to animal husbandry, fish farming, ecology, veterinary medicine and serves as a test to determine the level of iron concentration in fish muscles.

Iron is one of the most important chemical elements for all biological organisms, as well as for humans. The human body contains about 3.5-4.5 g of iron. 70% of iron is present in the blood, 30% - in the liver, bone marrow, spleen, muscles. Iron is involved in the transport and storage of oxygen, DNA synthesis, protein metabolism, the synthesis of thyroid hormones, the production of connective tissue, neurotransmitters, and plays an important role in maintaining immunity. However, this element is dangerous for the body in case of its excess, since there is a violation of carbohydrate metabolism, and cirrhosis of the liver and intestinal necrosis can also occur. Iron deficiency in the fish organism leads to growth retardation, the development of iron deficiency hypochromic anemia, a decrease in hemoglobin and hematocrit, as well as a decreased erythropoiesis.

Therefore, it is necessary to determine and control the level of iron in the body of animals, since its imbalance can cause a violation of the homeostasis of the body, as a consequence, affect the quality of the resulting product. Derivatives of leather are widely used to establish the concentration of heavy metals in organs and tissues of farm animals (Patent RU 2421726 C1, Patent RU 2426119 C1). In fish, a method for determining the copper content in muscle tissue using scales as a marker is known (Patent RU 2555518 C1).

Currently, there is a method for the determination of iron in fish (GOST 26928-86). Raw materials and food products. Methods for the determination of iron. - M: IPK Publishing house of standards, 2002. - S. 1-8.). However, this method does not allow for in vivo diagnostics of the level of iron concentration, since samples for analysis are already taken from killed fish. The method proposed by us differs from this method in that the chemical composition of scales taken from live fish is determined by the method of atomic emission spectrometry with inductively coupled plasma. Determine the concentration of Mg and / or Fe in the scales to determine the iron in the muscles of broodstock and females. And the regression equation is calculated:

y = - 0.0998x + 13.532, where x is the Fe content (mg / kg) in the scales,

y - the content of Fe (mg / kg) in the muscles.

y = - 1.943x + 12.918, where x is the Mg content (g / kg) in the scales,

y - the content of Fe (mg / kg) in the muscles.

This method can be used to estimate the iron content in the muscle tissue of fish of the species Stizostedion lucioperca. The problem is solved by establishing the level of concentration of Mg and / or Fe in the scales with further calculation of the level of iron in the muscles using the regression equations. According to the accumulation of optionally Fe and / or Mg in the scales, the iron content in the muscle tissue is determined.

An example of implementation. Scale samples were taken from pike perch (Stizostedion lucioperca) at the age of 3-4.5 years. The pike perch was caught between November and December 2019 in the Novosibirsk reservoir. The scales for this study were thoroughly washed. In order to clean the scales from contamination, a weighed portion of the scales was placed in a flask with distilled water, then the sample was stirred for one minute with a mixer at a rotation speed of 1000 rpm. Subsequently, the water was changed up to 10 times, repeating this stage. Then the scales were washed in acetone of the OSCh 49-5 brand for two minutes, then it was cleaned 3 times with deionized water and dried at room temperature.

For analysis, a 100 mg sample was weighed and placed in a quartz cup, then placed in a cold quartz oven. The furnace was heated to a temperature of 250 ° C, the exposure at this temperature was 15 minutes. Then the temperature was increased to 450 ° C with an exposure of 15 minutes. Then the sample was left in the oven to cool down to room temperature. After charring, the sample was ground in quartz cups, then a 10 mg sample was taken from the prepared sample powder and 50 mg of graphite powder and 40 mg of a spectroscopic buffer (15% NaCl and graphite powder) were added. For the study itself, 20 mg was taken from the resulting mixture.

Chemical elements in scales and muscle tissue were established using atomic emission spectral analysis on the basis of the Analytical Center for Collective Use of the Institute of Geology and Mineralogy. B.C. Sobolev SB RAS. To study the flakes, a two-jet arc plasmatron "Fakel" and a multichannel atomic emission spectrometer "Grand" manufactured by "VMK-optoelectronica" LLC (Russia) were used. To analyze muscle tissue, a Perkin Elmer 360 atomic absorption spectrophotometer (USA) was used. Samples for atomic absorption analysis were prepared in stages: after washing in soapy water, they were washed into the dishes with tap water, rinsed with bidistilled water, and then dried. Samples (weighing 100 g) were crushed to a homogeneous mass, then dried in an oven at a temperature of 60-70 ° C for about 12 hours until constant weight. From the prepared dry residue, 3 g were taken, which were ashed in a muffle furnace at a temperature of 500-550 ° C. Then, after 10-15 hours, the mineralization ended, the ash was gray or white. The samples were cooled at room temperature, then the resulting ash residue weighing 3 g was introduced into 3 ml of 50% hydrochloric acid, then the samples were heated on an electric stove to extract the dry precipitate, then this residue was transferred to a flask by diluting it in 25 ml of distilled water. The elemental composition was determined in the finished solution.

Data on the content of iron and magnesium in muscle tissue are presented in Table 1. In the muscles of pike-perch, the content of the trace element iron was many times less than that of the macronutrient magnesium. The phenotypic variability of iron was 1.8 times higher than the magnesium concentration (P <0.01).

Table 2 shows data on the content of magnesium and iron in fish scales. The iron concentration in the scales is 8.1 times higher than in the muscles (P <0.001). The individual variability of the iron content in muscle tissue and scales was equally high.

The variability of magnesium levels in scales was 1.5 times higher than in muscles (P <0.01). It was found that there are average correlation values between the studied indicators (Table 3). Therefore, if there are connections between these elements, it is possible to calculate the regression equations to predict the iron content in muscles by setting the concentration of one or more elements to choose from - Mg and / or Fe in the scales.

Therefore, using this method, it is possible to carry out an in vivo assessment of the interior of the fish by the iron content in the muscles, using only samples of scales.

Claims (5)

A method for determining the iron content in the muscle tissue of fish, including the analysis of a biosubstrate, characterized in that a microelement analysis of fish scales is carried out, and the concentration of one or more elements in the scales is detected at choice - Fe and / or Mg, and then the regression equations are calculated: y = - 0.0998x + 13.532, where x is the Fe content (mg / kg) in the scales, y - Fe content (mg / kg) in muscles, y = - 1.943x + 12.918, where x is the Mg content (g / kg) in the scales, y - the content of Fe (mg / kg) in the muscles.