SU1191789A1 - Method of determining bisnadine - Google Patents

Abstract

METHOD FOR DETERMINING VISNADINE by treating an analyzed sample with a chemical reagent followed by measuring the fluorescence intensity, characterized in that, in order to increase the sensitivity and selectivity of the determination and shorten the time of determination, an aqueous saturated bromine solution is used as the chemical reagent and the fluorescence intensity of the resulting solution is measured .

Description

00

about

The invention relates to analytical chemistry, and specifically to methods for determining visnadin, and can be applied for the qualitative and quantitative analysis of this compound in various materials for pharmaceutical research.

Visnadine is a derivative of furanochromone and is contained in the fruit of the plant ammisubly.

The aim of the invention is to increase the sensitivity and selectivity of the determination and shorten the detection time.

Example 1. Qualitative definition of visnadin.

8 to 10 ml of distilled water, 2 drops of a saturated aqueous solution of bromine are added to 1-2 mg of visnadin and shake. In ultraviolet light with a Ufa-3 or Ufa-6 light filter, strong violet fluorescence is observed.

Example 2. Qualitative determination of visnadin in vegetable raw materials.

1-2 g of the crushed raw material is extracted with 100 ml of a mixture of ethyl alcohol and chloroform (3: 7). To isolate visnadine, chromatography is applied on a thin, fixed layer of silica gel of brand LS 5/40 c. gypsum. The plates are activated at 110 ° C for 30 minutes. 0.1 mJ of the extract is applied to the plate and chromatographed in the solvent system hexane - ethyl ether-ethyl acetate (12: 12: 1). After that, the plate is dried and sprayed with an aqueous saturated solution of bromine. In UV light, a fluorescent bright violet spot is observed, which indicates the presence of visnadin.

Example 3. Quantitative determination of visnadin in the preparation “Avisan.

The Avisan tablets contain 0.05 g of the amount of furocoumarins and chromones isolated from ammonium one.

A fine weight (0.350 g) of finely divided Avisan tablets is transferred to a 100 ml volumetric flask, 20 ml of 96% ethanol is added, the mixture is shaken for 5 minutes and distilled water is added to the mark. Shake vigorously, drug solution, 5 - 10 min. This Avisan solution was transferred to a separatory funnel and extracted with 100 ml of chloroform, and the tide was added in portions of 10 ml. An aliquot portion of the chloroform extract (10 ml) was evaporated to dryness in a water bath and the residue was dissolved in 1 ml of ethyl alcohol (96%) and 8.9 ml of water. To the resulting solution, 0.1 ml of an aqueous saturated solution of bromine is added and the fluorescence intensity of the resulting solution is measured relative to the standard content of visnadin 10 µg / ml.

The content of visnadin is determined by calibration curve. The results of the determination of visnadin in Avisan are presented in Table. one.

Table 1

with

1 tablet of the drug contains 0.00041 g of visnadin, that is, about 8%. With this method of determination, the other compounds in solution with bromine do not fluoresce, which was tested experimentally. The completeness of the extraction (93-95%) was checked. chromatography of the solution of the preparation remaining after chloroform extraction.

Example 4. Quantitative determination of visnadin in the preparation of “Floverin.

Tablets "Floverin contain 0.05 g of visnadine and dihydrosamidine.

A precise weight (0.350 g) of finely divided Floverin tablets is transferred to a 100 ml volumetric flask, 20 ml of 96% ethyl alcohol is added, the mixture is shaken for 5 minutes, and distilled water is added to the mark. Shake vigorously for 5-10 minutes, the solution is the preparation. This solution of the preparation is transferred to a separatory funnel and extracted with 100 ml of chloroform, the tide is added in portions of 10 ml. An aliquot portion of the chloroform extract (25 ml) is evaporated to dryness in a water bath and the residue is dissolved in 0.2 ml of hexane. This amount is applied with a micropipette onto a chromatographic plate with a thin fixed layer of silica gel LS 5/40 / + 13% gypsum. Chromatographic in the solvent system hexane-isoamyl alcohol (2: 1) Rf visnadine is 0.58. The visnadin zone, detected by Rf size or standard visnadin sample, is quantitatively transferred to a test tube and 9.9 ml of distilled water are eluted. The completeness of the despair is 98-99%. To the solution, separated from the precipitate after elution, is added 0.1 ml of an aqueous saturated solution of bromine. Fluorimeter the test solution relative to the standard content

10 μg / ml visnadin. The content of the visnadin and the sample is determined by calibration curve. Tablets "Floverina contain about 48% visnadin.

Example 5. Quantitative determination of visnadin in ammonia.

5 g of finely ground fruit of the plant is extracted with 100 ml of a mixture of 96% ethanol and chloroform (3: 7). 0.1 ml of this solution is applied to a silica gel chromatogram of the brand LS 5/40 i + + 13% gypsum, activated for 30 minutes at 110 ° C. The coated sample plate is dried and placed in a hexane-diethyl ether-ethyl acetate (12: 12: 1) solvent system chamber and chromatographed using an ascending method. After drying

chromatograms for 5 minutes in air under ultraviolet light establish the localization of the visnadin zone in this zone, 38, noted by the visnadin standard, are quantitatively transferred into a test tube, 9.9 ml of distilled water are added, eluted for 5-10 minutes and, after separating the solution from the silica gel precipitate, add 0.1 ml of a saturated aqueous solution of bromine. Fluorimeter the resulting solution relative to the standard content of visnadin 10 µg / ml. The content of visnadin is determined by calibration curve. The fruit of the plant contains 0.008 g of visnadin, i.e. about 0.2%. Compounds of similar structure do not give such a reaction: coumarin, angelicin, psoralen, xanthotoxol, isopimpinelin, xantotoxin and others.

table 2

Pale blue

I rko purple

Pale blue, fast fading

In tab. 2 presents data on the fluorescence of visnadin and reliable samples of coumarins, observed when using the described method.

The fluorescence intensity of visnadin after its interaction with bromine does not change for 2 hours. The linear relationship between the intensity of flu-pale blue

Bright purple

Pale blue

orescence and concentration of visnadin is in the range of 0.5-10 μg / ml.

The analysis may be carried out with a fairly wide pH range in both neutral and acidic environments.

The sensitivity of the known method is 30 µg / ml, described by

0.5 µg / ml, which is 60 times higher than the known.

Using the described method is economical and requires less time (see table. 3).

Table 3

Preparation of mortar5

Chromatography

Chromatogram processing

Add reagents

Continued table. 3

Observation

The data show that the described method has a higher sensitivity, selectivity and expressivity than the known one and allows qualitatively and quantitatively to determine visnadine in a substance, medicinal preparations and vegetable raw materials. Possessing simplicity and cost-effectiveness, the proposed method is applicable in control-analytical laboratories and factory laboratories of pharmaceutical companies to control the quality of the drug and produce it, as well as in pharmacological studies for analyzing medicinal plants containing visnadine and searching for new plants - the sources of this compounds.

Claims (1)

METHOD FOR DETERMINING VISNADINE by treating the analyzed sample with a chemical reagent and then measuring the fluorescence intensity, characterized in that, in order to increase the sensitivity and selectivity of the determination and reduce the determination time, an aqueous saturated bromine solution is used as a chemical reagent and the fluorescence intensity of the resulting solution is measured.