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Gabunia Ketevani, Akaki Tsereteli state university, PhD., Pharmacy, the faculty of Medicine, Kutaisi E-mail: ketevanigabunia@gmail.com Abuladze Nino,
Akaki Tsereteli state university, PhD., Pharmacy, the faculty of Medicine, Kutaisi E-mail: nino.abuladze5@atsu.edu.ge
DETERMINING THE CONTENT OF TANNING SUBSTANCES IN THE ANTIMYCOTIC OINTMENT BY METHOD OF UV SPECTROPHOTOMETRY
Abstract: Despite the wide range of modern medications, mycoses remain one of the most common diseases in the population, with a growth each year in the number of people suffering from this disease. While studying folk manuscripts and mediaeval medical handbooks, we have discovered interesting materials about the properties of plants containing tanning substances. It has been revealed that medicinal plants containing tanning substances exhibit antimycotic activity. Tanning substances abound in the nature of Georgia. They are contained in medicinal plants, fruits and vegetables. On the basis of a literary review we conducted research and prepared ointment. Microbiological studies of the ointment have confirmed that medicinal plants containing tanning substances exhibit antimycotic activity. The ointment has exhibited a high antimycotic effect, and its studies are still going on. The existing standard and the normative documentation we have developed specified determining the content of tanning substances in the ointment by titration, in particular by the method of permanganometric titration. This is a labor-intensive and time-consuming method. Based on the study of data from literature, we made it necessary to modernize the spectrophotometry method for determining the content of tanning substances in medicinal plant raw materials for our antimycotic ointment.
The method of UV spectrophotometry that we have developed is effective, sensitive and simple as compared with the method of permanganometric titration.
Keywords: UV spectrophotometry, antimycotic ointment, polyphenolic, tanning substances.
Introduction esis inhibition, and antiviral properties. Tanning substances
Despite the wide range of modern medications, mycoses abound in the nature of Georgia. They are contained in medic-
remain one of the most common diseases in the population, inal plants, fruits and vegetables. On the basis of a literature re-
with a growth each year in the number of people suffering from view, we have undertaken studies and developed the ointment.
this disease. According to World Health Organization, approxi- Microbiological studies of the ointment have confirmed that
mately 20% of the world population suffers from mycosis infec- medicinal plants containing tanning substances exhibit anti-
tions and the percentage is even higher in tropical and subtropi- mycotic activity [3,77-81; 4,564; 5,179; 6,67-81; 7,240-241;
cal areas [1]. Scientists believe that, along with other reasons, 8,205-215]. The ointment was developed within the grant
this is caused by uncontrolled use of antibiotics, self-treatment for fundamental researches from Shota Rustaveli Georgian
with antimycotic preparations ofvarious groups, which results National Science Foundation FR/28 /8-403/13, and its
in lowering of human immunity and a further health complica- standardization method was also developed. By this method,
tion [2,249-252]. In addition, despite the diversity of antimy- we determined the total content of tanning substances in this
cotic drugs, their use frequently does not lead to full recovery ointment, which is estimated at no less than 10%. from this disease the elimination of a relapse. The ointment has exhibited a high antimycotic effect, and
While studying folk manuscripts and mediaeval medical its studies are still going on. The existing standard and the nor-
handbooks, we have discovered interesting material about the mative documentation we have developed specified determin-
properties of plants containing tanning substances. Tanning ing the content of tanning substances in the ointment by titra-
substances, tannins are the most common polyphenolic com- tion, in particular by the method of permanganometric titration
pounds with a bitter taste, which are characterized by the high [9]. This is a labor-intensive and time-consuming method. It
biological and pharmacological effects, such as carcinogen- had also become necessary to improve the existing standard.
DETERMINING THE CONTENT OF TANNING SUBSTANCES IN THE ANTIMYCOTIC OINTMENT BY METHOD OF UV SPECTROPHOTOMETRY
The European Pharmacopoeia describes the qualitative and quantitative determination of tannins in eight plant species containing tanning substances by simple colored reactions, using microscopic, chromato-detector and chemomet-ric methods [10, 24].
Based on the study of data from literature [11, 413-415; 12,190-193], we made it necessary to modernize the spectrophotometry method for determining the content of tanning substances in medicinal plant raw materials for our an-timycotic ointment, for which we produced validation of the ultra-violet spectrophotometric method for determining the content of tanning substances in medicinal plant raw materials [13, 176]. A comparison of the results obtained by spectrophotometry method was carried out by method of perman-ganometric titration.
Research subject and method
Research subject was 10 g of a multi-component ointment. The base components per 10 g of base: distillated monoglycerides - 1.0 g; glyceril mono stearate - 4.0 g; glycerin - 3.0 g; cetyl palmitate - 2.0 g; emulsifying wax - 6.0 g; hostacerine - 2.0 g; stearin - 6.0 g; paraffinic (perfume) oil -7.0 g; cacao oil - 6.0 g, sodium benzoate - 0.3 g; a 16% solution of dimexidum - 4.0 g; purified water - up to 100 g. 100 g of ointment contain 84.0 of base component and 16 grams of therapeutic agents, including plant extracts (the dense extracts of nut Corylus avellanta and tannin Cotinus Coggygria) and 40% lactic acid, so that the total tannin content in the preparation should be no less than 10%.
Research method: UV spectrophotometry. The quantitative content of tanning substances was determined using a "Shimadzu-UV-240" spectrophotometer.
Experimental part
Preparing experimental sample solution: we placed 2.0 g of ointment under study in the flask of 50 ml capacity, adding 10 ml of distilled water, heated on water bath by boiling for 3 minutes, and then we cooled and filtered it in a 100-ml measuring flask with ground-stopper. We carried out this process three times. At the end, we washed the filter in 5 ml of hot distilled water 6 times until the negative reaction to tannin. After binding the obtained extracts, the volume of the flask was brought up to the mark with distilled water.
Preparing tanning standard sample solution: 0.0025 g of tannin standard sample reduced to a constant weight (100°-105° C), we placed in the flask of 50 ml capacity. With a Table 1.- The quantitative content of
continuous mix, we added 50 ml of 70% ethyl alcohol. 2.5 ml of the obtained solution 25 we placed in a 25-ml measuring flask, the volume of the flask was brought up to the mark with 70% ethyl alcohol.
Results and their discussion
5 ml ofwatery extract obtained from material under study (ointment), we placed in a 50-ml measuring flask and added 70% ethyl alcohol up to the mark. 2.5 ml of this solution, we transferred into a 25-ml measuring flask and the volume was brought up to the mark with 70% ethyl alcohol. The optical density of the obtained solution was determined by a spectrophotometer "Shimadzu-UV-240" at a wavelength of275 nm as regard to 70% ethyl alcohol. The thickness of a sample holder layer is 10 mm. Simultaneously, we determined the optical density of a standard sample of tannin under the same conditions.
The total content of tanning substances was calculated by the formula:
where
, D x Mix 50 x 25 x 2,5
x % =-— x 100%
Di x M x 5 x 50 x 2,5
D - the optical density ofa solution under study (ointment);
D1 - the optical density of a standard sample of tannin;
M - the weight of an ointment under study, g;
M1 - the weight of a standard sample of tannin, g.
At the beginning of the research, we studied the quantitative content of tanning substances in the antimycotic ointment by the methods of UV spectrophotometry and perman-ganometric titration. A comparison of the results obtained showed that both methods had yielded almost the same results (by I method - 10.29%, and by II method - 10.25%). thereafter, the studies were carried out by the method of UV spectrophotometry.
The research was conducted for the period of six months at certain intervals: 3-4 days a week. Each day, we ran a test 3 times. To conduct a study, we used the ointment prepared at the beginning of the research (for the period of 6 months) and ointment samples prepared each week.
Statistical processing of the data obtained showed that the quantitative content of tanning substances in the ointment samples prepared during the period of 6 months and in samples prepared each week, are identical, and complies with the requirement of the existing regulatory and technical documentation (standard - not less than 10%). The quantitative content of tanning substances in samples is given in (Table 1).
tanning substances in the ointment
Sample The content of tanning substances,% The average amount
I month II month III month IV month V month VI month
Sample 1 10.2 10.15 10.15 10.15 10.2 10.15 10.17
Sample 2 10.2 10.1 10.3 10.1 10.2 10.1 10.17
Sample 1 - the ointment first-prepared during the period of 6 months.
Sample 2 - the ointment prepared each week.
The table shows that the percentage of tanning substances in the ointment prepared during the period of 6 months remains unchanged that is the indicator of its state.
Conclusions
Based on the data obtained, we can conclude that:
1. To determine the quantitative content of tanning substances in the antimycotic ointment, it is possible to use method of UV spectrophotometry;
2. The method of UV spectrophotometry is effective, sensitive and simple as compared with method of perman-ganometric titration.
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