CC BY
8
https://doi.org/10.29013/AJT-23-3.4-47-52
Olimov Khairullo Kayumovich, Candidate of Pharmaceutical Sciences, Associate Professor,
Tashkent Pharmaceutical Institute Kholikova Zukhra Akhmadovna, Assistant, Tashkent Pharmaceutical Institute
Abadjanov Damir Iserkepovich, Assistant, Medical Institute of the Republic ofKarakalpakstan
IR-SPECTROSCOPIC METHOD TO HELP IN CHEMICAL-TOXICOLOGICAL INVESTIGATION OF AMLODIPINE
Abstract. Diseases of the cardiovascular system are very common among the population today. According to statistics, 17 million people live in the world every year. One of the main reasons for this is acute heart rhythm disturbances, problems with high blood pressure. In particular, ventricular tachycardia and fibrillation of cardiac arrhythmias are very life-threatening. Therefore, in connection with life -threatening severe rhythm changes, such as tachycardia and irreversible ventricular fibrillation, the development of suitable antiarrhythmic drugs that can be fast and highly effective even in severe arrhythmias remains one of the urgent tasks of today. Based on this, the relevance of our work is the development of methods for separating amlodipine from a biological object, improvement of methods for analyzing the drug amlodipine, enalapril and metaprolol which is widely used in the treatment of hypertensive diseases.
Keywords: hypertension, biological object, substance, tablet, drug, blood, urine, methods of analysis, solvent system.
Actuality of theme. Diseases of the cardiovascular system are very common among the population today. According to statistics, 17 million people live in the world every year (Oshchepkova E. V., 2007). One of the main reasons for this is acute heart rhythm disturbances, problems with high blood pressure. In particular, ventricular tachycardia and fibrillation of cardiac arrhythmias are very life -threatening (Wa-then M. S., 2004; Anthony R., 2008; Knecht S., 2009; Juan J. S.M., 2010). Therefore, in connection with life-threatening severe rhythm changes, such as tachycardia and irreversible ventricular fibrillation, the development of suitable antiarrhythmic drugs that can be fast and highly effective even in severe arrhythmias remains one of the urgent tasks of today. Based on this, the relevance of our work is the
development of methods for separating amlodipine from a biological object, improvement of methods for analyzing the drug amlodipine, enalapril and metaprolol which is widely used in the treatment of hypertensive diseases.
Communication of dissertation work with research plans. Practise an assay method that separates the drug from the biological object, causing
hypersensitivity reactions to the drug amlodipine, which lowers blood pressure and normalizes heart function in hypertensive patients.
The purpose of the study.
1. Development of hypersensitive chemical reactions to antihypertensive drugs;
2. To determine the sensitivity of the reaction to antihypertensive drugs;
3. Improvement of methods of quantitative analysis of antihypertensive drugs;
4. Development of a method for separating antihypertensive drugs from biological fluids and biological objects.
Object and subject of research. Antihypertensive drugs that lower blood pressure.
Research methods: during the analysis, the use of chemical reactions, the creation of qualitative reactions to the preparations obtained for the analysis, the use of thin layer chromatography, spectrophotometry and high-efficiency liquid chromatography methods, as well as the use of infrared spectroscopic spectroscopy methods.
Scientific, theoretical and practical significance of the research results. Implementation of the developed methods of analysis in the Republican laboratories of forensic chemistry.
Introduction
Amlodipine is a derivative of digiropyridine and has antianginal and hypotensive effects. At the same time, it causes a number of unpleasant reactions, and in some cases cases of poisoning have also been observed. As a result of excessive vasodilatation, there is a sharp drop in arterial pressure and tachycardia.
CHClNO. C6H6O3S mm=567.1
20 25 2 5 6 6 3
(RS)-2-(2-aminoethoxymethyl)-6-methyl 4-(2-chlorophenyl)-1,4-dihydropyridine - benzene-sulfonate of 3,5-dicarboxylic acid.
Amlodipine is widely used in medical practice as a hypotensive agent. In some cases, acute poisoning occurs when it is taken in excess of the norm. In such cases, the patient's urine and blood are analyzed and a conclusion is drawn. Determination of the authenticity of the substance was carried out by the method of thin layer chromatography. When amlodipine is isolated from biological liquid urine and determined
by TLC method, 2 g of ammonium sulfate is added to 15 ml of urine containing 10 mg of the substance, extracted 3 times (5 ml) with chloroform, 1 g of anhydrous sulfate is filtered through a filter containing sodium sulfate, and washed with 2 ml of chloroform. The chloroform was evaporated to a dry residue. The residue was dissolved in 2 ml of alcohol, then the solution was dripped onto the starting line of a silufol UV-254 plate with a diameter of 3 mm using a capillary and dried. Then 1 mol was chromatographed in a chromatographic chamber saturated with acetic acid vapor. When the plate was dried from the chamber and exposed to iodine vapor, a blue spot with Rf=0.87 was observed. At the same time, an IR spectroscopic method for determining the authenticity and quantity ofthe substance extracted from the biological fluid by the method described above was developed.
Basic phrases. Amlodipine, thin layer chromatography, silufol - UV-254, IR -spectroscopy, valence and deformation vibrations, intensity of absorption path, validation.
The purpose of the study. Chemical-toxicologi-cal studies of amlodipine are of great importance, allowing quick and accurate investigation of cases of poisoning. We aimed to develop an infrared spectroscopic method for the extraction and analysis of amlodipine from biological fluids.
Practical part. From the IR spectrum of amlodipine isolated from biological fluid, the following classification absorption pathways were determined: n max: cm -1, 3301 primary NH2 group, 3190 (pyri-dine ring N-H valence vibrations amino group valence vibrations), 2981 (C-H valence vibrations in ethyl and methyl groups); 2948 (valence vibrations of C-H bonds in aromatic ring and pyridine ring) 1698, 1675 (valence vibrations of carbonyl bonds in carboe-thoxyl group, 1615 (valence vibrations of aromatic ring - C=C bonds), 1493 (valence vibrations ofC=C bonds in the pyridine ring), 1445, 1432 (deformation vibrations of methyl groups), 1384, 1365 (deformation vibrations of C-H groups in the aromatic ring and pyridine ring), 1303, 1265 (out-of-surface defor-
mation vibrations ofC-H groups), 1202, 1125 (plane deformation vibrations of C=C bonds to aromatics), 1095, 1034, 1049 (carboxyl and oxyethiamine and categories C-O-), (1,2, Plane strain vibrations of 4,5-substituted pyridine C=C bonds, 869, 838, 791 (C-H bonds of 1, 2, 3 substituted aromatic rings), 735, 615 cm-1 (C-H valence vibrations).
Quantitative analysis. Infrared spectra of a 1 mg (exact draw) amlodipine working standard sample and samples isolated from a biological object were obtained in the form of pressed KBr tablets, and the areas of carboethoxyl groups were measured as characteristic absorption pathways.
Figure 1. - IR spectrum of amlodipine isolated from biological fluid
Figure 2. IR-spectrum of a working standard sample of amlodipine
The percentage of amlodipine extracted from the in the formula: S - the area of absorption paths of car-
biological object was determined using the following bonyl groups in the IR spectrum ofthe standard sample;
calculation formula. S0 - amlodipine working standard sample IR -
S-m ■ P-100 area of absorption paths of carbonyl groups in the
& -m-100 ' spectrum;
X = -
m - Drawer of amlodipine isolated from a bio- P - amlodipine is the percentage of amlodipine logical object; in the working standard sample.
m0 - drawer of the woing standard sample of amlodipine;
Metrological description and validation of the analytical method
Table 1.
№ m m0 X X S2 S Ax A X E% E % RSD% v%
1 0.00090 0.00010 98.4
2 0.00095 98.7
3 0.00098 99.2 99.2 0.32 0.57 1.57 0.7 1.71 0.76 0.6 0.32
4 0.00103 99.6
5 0.00107 100.2
Specificity of the method. When the spectra obtained at different concentrations and with different equipment are compared, the intensity and classification values of the absorption lines in them are the same, and the absorption lines of the carbonyl groups in the spectra were observed at 1698, 1675 cm-1.
Linearity of the method. The linear relationship was determined in the dimensions of the relationship between the draw and the characteristic absorp-
tion path for obtaining an IR-spectrum, the correlation coefficient was 0.9992 nm, and the graph of the linear relationship was found to be y = 3.5833 x x - 0.0167.
IR-spectrometric quantitative analysis method of amlodipine linearity of the relationship between the amount of substance and the area of the absorption path
Figure 3.
The precision of the method was determined in six re-analyses and was determined by studying the
metrological description of the results.
Table 3.
X g X% X - X, d2 Metrological description
1 2 3 4 5
0.00090 98.4 1.2 1.44 S2 = 0.75 AX = 0.69
0.00095 98.7 0.5 0.25 S = 0.27 A X=0.28
1 2 3 4 5
0.00098 99.2 0 0 SX = 0.11 E = 0.27
0.00104 99.6 0.4 0.16 E=0.12 V=0.75
0.00108 100.2 1.0 1.0 RSD=0.8%
The reproducibility of the method was carried experiments, and processing the obtained values out by analyzing the samples extracted from the with the method of mathematical statistics. biological fluid six times on the basis of triplicate
Table 4.
Return № number Percentage of Amlodipine
Experiment 1 Experiment 2 Experiment 3
1 98.4 98.1 98.2
2 98.7 98.8 98.6
3 99.2 99.4 99.8
4 99.6 99.7 100.2
5 100.2 100.5 100.8
6 100.5 101.4 101.8
RSD 0.8 2.1 1.8
The accuracy of the developed method was determined by comparing the results of experiments repeated 9 times in three series in samples with 10%: 20%: 30% more than the nominal value
Table 5. - Determining the accuracy of the equipment
Amlodipine drawer, g Amount added, g Total amount, g Determined amount, g Determined amount,%
0.001 0.0001 0.0011 0.0009 82
0.001 0.0001 0.0011 0.0009 82 >88
0.001 0.0001 0.0011 0.0009 100
0.001 0.0002 0.0012 0.0011 92 ~
0.001 0.0002 0.0012 0.0010 83 92.6
0.001 0.0002 0.0012 0.0012 100^
0.001 0.0003 0.0013 0.0011 85
0.001 0.0003 0.0013 0.00128 98.5 >94.5
0.001 0.0003 0.0013 0.0013 100
Average value 91.7%; The range of values is 88-94.5%
Conclusion 2. The isolated amlodipine was identified using
1. A moderate method of extracting amlodipine chromatographic and physical methods;
from biological fluid was developed. The most suit- 3. The absorption pathways of the IR-spectrum
able extractant was found to be chloroform; of amlodipine were determined;
4. IR-spectroscopic method of quantitative analysis for amlodipine was developed;
5. The method was validated according to param eters such as precision, accuracy, reproducibility, lin earity.
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