Method for predicting body intoxication with lead

FIELD: medicine, diagnostics.

SUBSTANCE: due to spectrophotometry on should study body biological material to detect the concentration of porphyrin in tissues of biological material and compare it values against the standard. As biological material one should apply that part of the tissue at patient's body where spectrophotometric investigation should be carried out in vivo. Moreover, One should affect this part with low-intensity laser radiation, wave length corresponds to one of the maximums of protoporphyrins' absorption range. One should determine the amplitude of fluorescence radiation of this part. As a standard for comparison one should take the amplitude of fluorescence of protoporphyrin IX that models the concentration of protoporphyrin in blood erythrocytes ranged 100-1500 mcg/l and more. At equality of amplitudes it is necessary to establish concentration of proptoporphyrin IX in blood erythrocytes of biological tissue under testing. Moreover, wave lengths corresponding to maximums of absorption range of protoporphyrins correspond to 337, 370, 405, 532, 632 nm. The innovation enables to considerably simplify the investigation of values that indirectly characterize lead concentration in tissues and, thus, total degree lead intoxication in patients with the risk of lead intoxication.

EFFECT: higher efficiency of diagnostics.

1 cl, 1 ex

 

The invention relates to medicine, namely to methods for the rapid diagnosis of lead intoxication of living biological tissues and the whole organism, and can be used in pathology, toxicology, therapy, and other fields of medicine.

Known methods of determining the concentration of lead in blood and urine, as well as indicators of its impact (coproporphyrin and Delta-aminolevulinic acid) laboratory methods. For example, one method for determination of lead in blood and urine is dry ashing of samples of blood and urine, its dissolution in indifferent electrolyte and subsequent polarographic determination of lead (see "Polarographic determination microgramme quantities of lead in the blood" - Pavlovskaya N.A., etc. / Laboratory business, No. 1, 1982. - p.26-29). For the assay, 1 g of blood and 20 ml of urine with heparin, produce mineralization biosubstrates with sulfuric and nitric acids followed by obtaining the dry residue when heated tile with asbestos gasket and further combustion in a muffle furnace. Solemny the remainder of the blood is diluted hydrochloric acid, and urine background solution and produce polarographic analysis, pre-equalising researched solutions with an inert gas to remove samples of oxidants. Before each series of polarographic analysis should be done to the control sample for the presence of lead in the reagents.

There is a method of assessing the severity of lead poisoning, including spectrophotometric study of the biological material of the organism (Lyubchenko PN. Features of the clinical examination of workers in lead industries / guidelines. - M.: MONICA, 1985.-22 C.).

This produces a definition of the levels of accumulation of coproporphyrin and Delta-aminolevulinic acid in the urine.

In the practice of clinical laboratory analysis to determine coproporphyrin method is often used in Fischer modification Migunova and Ukimerioni. Extraction of coproporphyrin of urine is carried out with ether. The mixture of urine, acetic acid and ether is shaken on shuttleport, separate the aqueous and ether phases. Next remove coproporphyrin from the ether by adding hydrochloric acid and shaking for 5 min test tubes, leaking hydrochloric acid in a measuring tube, repeating this operation 4-5 or more times (until complete discoloration layer of hydrochloric acid). The resulting solution photometrate on the spectrophotometer.

Determination of Delta-aminolevulinic acid in urine is produced using ion exchange resins. The upper layer of the resin cover filter paper, miss distilled water, then with 1 ml of urine, which passes through the resin in the glass, leaving the ion exchanger urea and Elita-aminolevulinate acid. Urea is removed by passing water through the resin. Further isolated from the resin of the Delta-aminolevulinate acid using sodium acetate and sodium acetate buffer. The resulting solution also photometrate on the spectrophotometer.

The main disadvantages of these methods are:

1) the difficulty and complexity of techniques, long duration of time, which makes the frequent repetition of research in monitoring to determine the effectiveness of therapy;

2) the inability to use methods for screening during the examination of workers in lead industries directly in a production environment;

3) study of lead content in the blood is invasive methods, which is particularly undesirable given the increasing incidence of viral hepatitis and other diseases transmitted through blood.

The problem posed by the authors, is to eliminate these drawbacks by developing a simple, cheap, non-invasive diagnostic method, based on modern technology fluorescence spectrophotometric analysis in vivo.

In addition, the objective was also to enhance the functionality of the method by determining the increased local concentration of protoporphyrin on the tissue under normal ur the outside concentration of lead in blood and urine and to assess the severity of the local lead intoxication tissue that will provide the appropriate treatment.

This task is achieved in that in the method for the diagnosis of lead intoxication, including spectrophotometric study of the biological material of the organism, the concentration of porphyrin in the tissues of a biological material and a comparison with the standard proposed as a biological material to use piece of tissue on the patient's body on which to conduct in vivo spectrophotometric study, to affect the area of low-intensity laser radiation with a wavelength corresponding to one of the maxima of the absorption spectrum of protoporphyrin, to determine the amplitude of the fluorescence radiation of this section, and as a reference for comparison to take the amplitude of the fluorescence of protoporphyrin 1X modeling the concentration of protoporphyrin in red blood cells from 100 to 1500 µg/l or more, and in case of equal amplitudes to establish the concentration of protoporphyrin 1X in the red blood cells of the investigated biological tissue.

Moreover, the wavelengths corresponding to the maxima of the absorption spectrum of porphyrins form 337, 370, 405, 532, and 632 nm.

It is known that porphyrins and their various chemical compounds and derivatives have strong fluorescence spectra in the wavelength range of 600-800 nm (Judenfrei C. Fluorescence analysis is in biology and medicine. - M.: Mir, 1965). The excitation of the fluorescence of porphyrins called their illumination light with a wavelength corresponding to one of the maxima of the absorption spectrum of porphyrins(337, 370, 405, 532, 632 nm). It is also known that such biological tissue, such as skin and mucous membranes of the organs are for optical radiation translucent and light-scattering environments (Tuchin VV Lasers and fiber optics in biomedical research. - Saratov: Saratov state University, 1998). Therefore, when the illumination of the biological tissue by optical radiation it penetrates the tissues, interacts with its biochemical and morphological structure and due to the acts of multiple scattering partially emerges from the illuminated surface, forming the so-called secondary flow of the scattered radiation from the surface. If within the biological tissue contains fluorescent substances such as porphyrins, together with the flow of secondary scattered radiation from biological tissue will be issued and the flux of the fluorescence of these substances. This thread will be different from the stream usually scattered radiation its characteristic spectrum and intensity. The emission spectrum of the fluorescence will indicate which fluorescent substances caused the fluorescence and the intensity of fluorescence will correlate to what iCustom fluorescent substances, contained in the volume of tissue exposed to light.

In order for the intensity of fluorescence was possible in vivo to quantify the level of accumulation of fluorescent substances in the tissue (in this case, porphyrins), you spectrophotometric apparatus to register the signal amplitude of the fluorescence of porphyrins living biological tissue and to compare the amplitude of any of the reference values of the amplitudes of signals corresponding to the different levels of accumulation of porphyrins in biological tissue.

The method is as follows.

In the selected tissue of the patient in vivo carry out spectrophotometric study. This affects the area of low-intensity laser radiation with a wavelength corresponding to one of the maxima of the absorption spectrum of porphyrins (337, 370, 405, 532 or 632 nm)log amplitude emerging from the biological tissue fluorescence radiation and compare it with a reference amplitude of the fluorescence of protoporphyrin 1X in the concentration range of 100-1500 mg/l and >1500 µg/L. In case of equality of these amplitudes establish the concentration of protoporphyrin in red blood cells of the investigated biological tissue - Par. Then, in accordance with generally accepted grading the severity of lead intoxication (Ammonemia, Ogur the powa Nsarchive and other Clinic, diagnostics, treatment, examination of disability and prevention of lead poisoning / guidelines - M.: Russian Ministry of health, 1986. - 25 C.) determine the severity of lead intoxication:

- initial (100 μg/l<Par.<500 µg/l)

- lightweight (500 g/l<Par.<1500 µg/l)

and expressed (Per.> 1500 µg/l)

where PPAR. the concentration of protoporphyrin in red blood cells.

A specific example of the method.

Patient I., 1958 born., and/b No. 16007, was in the Department of pathology MONICA with 10.09. on 16.10.2003, was Admitted with complaints of General weakness, pain in the arms, leg muscles, limitation of movements, abdominal pain, fever. 1994 and currently works at the Podolsk battery plant collector-palsecam. The duties of palika includes soldering interelement connections battery lead rods using a gas burner when the movement of lead batteries in the pipeline. Harmful production factors in the work of palika are lead and its inorganic compounds, noise. The duration time of exposure per shift is 83%. The concentration of lead in air of the working area in different years was 0,307-1.5 mg/m3MPC 0.01 mg/m3(the exceedance in 30-40-55 times). Since 1997 has been a marked increase in Delta-ALA (50-80-94 µmol/l), appeared above the written complaint, the strengthen which in August 2003, was the reason for hospitalization in the Department of pathology MONICA. Objectively: General condition of moderate severity. Height 172 see Weight 50 kg Skin and visible mucous membranes pale. In the mouth a lot of artificial and broken teeth, crowns, teeth - stones that lead the border could not be clearly defined. The strength in the muscles of the shoulder girdle is sharply reduced. In the lungs vesicular breathing, wheezing is not heard. BH 18 in 1 min muffled heart sounds, acadeny, HR 100 in 1 min. HELL 110/70 mm Hg Tongue coated white bloom. Abdomen palpation soft, painless. The liver is not enlarged. With the first tapping negative on both sides. Analyses: 1) blood Lead 110 µmol/PP (rule 40). 2) Lead in urine 278 μmol/% (normal is 40). 3) Coproporphyrin in the urine 1939,2 µg/g creatinine (normal range 20-80). 4) Delta-ALA in urine 31,5 mg/g creatinine (normal 2.5). Determining the severity of lead intoxication proposed by the fluorescence spectrophotometry in vivo on the skin of the fingers showed that the amplitude of the fluorescence signal is in the range of concentrations of protoporphyrin in erythrocytes from 500 to 1500 µg/l, which corresponds to mild degree of intoxication.

The use of this method in clinical practice will significantly simplify the study of indicators, indirectly characterizes the x concentration of lead in the tissues, consequently, the overall severity of lead intoxication in individuals with suspected lead poisoning. This method allows in the shortest time period necessary to conduct dynamic monitoring of patients with lead intoxication when assigning them etiotropic and pathogenetic therapy, in addition, possible mass screening in the production conditions during the preventive medical examinations. The proposed method also allows you to explore the increased local concentration of protoporphyrin in the tissues of the normal level of lead concentration in blood and urine and to assess the severity of the local lead intoxication tissues.

1. Method for the diagnosis of lead intoxication, including spectrophotometric study of the biological material of the organism, the concentration of porphyrin in the tissues of the biological material and comparing its value with the reference value, characterized in that the biological material used piece of tissue on the patient's body, in which in vivo carry out spectrophotometric study, thus affecting the area of low-intensity laser radiation with a wavelength corresponding to one of the maxima of the absorption spectrum of protoporphyrin determine the amplitude of the radiation fluorescence the this site, but as the standard of comparison take the amplitude of the fluorescence of protoporphyrin IX, which simulates the concentration of protoporphyrin in red blood cells from 100-1500 mg/l or more, and in case of equal amplitudes establish the concentration of protoporphyrin IX in erythrocytes of the blood of the studied biological tissue.

2. Method for the diagnosis of lead intoxication according to claim 1, characterized in that the wavelengths corresponding to the maxima of the absorption spectrum of protoporphyrin, form 337, 370, 405, 532, and 632 nm.



 

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