CS228259B1 - Method for determining lobeline chloride content in countersmoking chewing-gums - Google Patents

Description

The invention relates to a method for determining lobelin chloride in anti-smoking chewing gum.

Anti-smoking therapies rely on psychotherapeutic efficacy, often supplemented by the addition of ataractics, or by the application of substances that, in smoking, induce unpleasant taste and olfactory sensations and thereby induce resistance to smoking. For this purpose, they are using pooled salt products containing lobelin as a smoking cessation agent or a smoking cessation agent. This substance has similar pharmacological effects to nicotine. Both alkaloids stimulate the respiratory center. They are able to stimulate the release of oerotonin after injection. Typical of both alkaloids is their ability to release catecholamines from adrenal pulp. Joot-Jochim's Lobelin - Nicotine Syndrome Joot J., Jochim K .: Med. Kline. 54, 1049 (1959). This syndrome consists in the fact that smoking after previous oral administration of lobelin gives rise to unpleasant side effects such as dizziness, headache, vomiting irritation, severe sweating and pain in the ordca area, which affects resistance and contributes to quitting smoking. Lobelin oa is administered orally in the form of a salt, such as lobelin sulphate or hydrochloride. It is applied oa in solid dosage forms in the form of tablets, palatines and chewing gum.

The composition of all three types of dosage forms differs from each other only by the auxiliaries used and the manner of their galenic processing.

The first expert report on the manufacture of lobelin for its use as an anti-smoking ingredient was prepared by Urban J. Lobelin. Study report O.S.VTEI farm.prum.

Several authors gave analytical evaluation of lobelin chloride o and o. Its chewing in chewing gums encountered considerable tensile bones for lobelin thermolability, which complicated its extraction from the chewing gum. Defining as the fact that one of the lobeline chloride separanda not ^j: E may be used, protifajčiaroke gum bezjpresného determining the concentration of the dosage form.

Polarographic determination of lobelin chloride proposed by Šautavý, Šantavý F .: Čeo.čeo.lek.56, 1, 1943 and Šantavý F .: Čes. 57, 109, 1944 could not be accomplished with the aforementioned demanding chewing gum extraction and lobelin thermolability. The quantitative separation of lobelin chloride from the ottotic bags of Chewing gum could not be transferred without decomposition and therefore the results of the polarographic assay were not reproducible.

The photometric method used to determine lobelin chloride after its conversion to acetophenone based on the 2,4-dinitrophenylhydrazine color reaction described by Martern K., Schill C .: Svenok.farmac.Tidskr.54, 461, 1950 could not be used intensively the coloring resulting from the reaction of the chewing gum mass with this reagent, so this procedure was not suitable for the eye of interest.

On the basis of the above, it appeared necessary to develop a method which would allow reproducible determination of lobelin chloride in anti-smoking chewing gums, which was not possible and would also negatively affect its use in anti-smoking counseling. The chewing gum used for this sack contains, besides lobelin salt, as adjuvants, 3-orbitol, propylene glycol, chewing gum mass and as a taste corrected sugar or milk sugar. In developing the analytical procedure, the objective was to arrive at a method that would give exact and reproducible assay results.

A method for the determination of lobelin chloride in anti-smoking chewing gum based on its conversion to acetophenone was prepared and the acetophenone was isolated by distillation and determined by liquid chromatography at 20 ° C and 12 MPa using a chemically bound octadecyloilane-size silica gel 10 µm grains and as a solvent solvent methanol-water, at a ratio of 1 2.

Chewing gums intended to prepare the calibration curve are converted to acetophenone similar to that of chewing gums in which

The 4,228 2S9 has a thaw of 3 and after isolation of the acetophenone formed, this is determined by liquid chromatography. The results of the assay in this concentration range are reproducible, allowing analysis of the anti-smoking chewing gums and hence their preparation, and safe use in anti-smoking counseling centers, which was previously not possible.

table

Experimental results r «calculation for construction of calibration curve / x / '

Lobelin chloride concentration in chewing gum Average values of y from values of y in integration units. Values from the regreece equation y 0.03 6676 6673 0,045 9475 9257 0,060 12274 12359 0,075 15073 15778 0,090 17872 17287

/ x / Estimation of the standard error of the regression line o = 546 Correlation coefficient r _VY = 0,994

The calibration curve equation below was calculated from the measured values by linear regression.

y = 1077 + 186611 x, where y is the area of the chromatographic peak in integration units, x is the reported amount of lobelin chloride in 100 g of chewing gum. The results are summarized in the table.

In FIG. 1 is a calibration curve for the determination of lobelin chloride.

In FIG. 2 is a chromatogram of a gum of lobelinium chloride with a taste coefficient for sucrose and sorbitol.

In FIG. 3 is a chromatogram of a chewing gum containing lobelin chloride with flavoring lozenges and sorbitol. , _jt

ΑλΛτ- t / e eAx "iue / ear" (M funtffcy ".fc / aiezv eá / .r / efa / dwAiyk"

In FIG. 1 on the x-axis shows the weight of lobelin chloride in 100 g of chewing gum. The y-axis shows the area of the curve in integration units. line 1 represents the dependence of the area of the chromatographic wave on the concentration of lobelin chloride in the chewing gum, the area being given in integration units. The value given is the average of the six measurements. Line 2 shows the regression curve calculated

- 5 linear regression from all measurements. 23i 258

In FIG. 2 elution wave 3 is acetophenone chromatographic wave. Elution waves 4 and 5 are the chromatographic waves of the components of the disintegrant mesh.

In FIG. 3 is a chromatogram of chewing gum containing lobelin chloride with flavoring agents lactose and aorbitol. Elution wave 3 is an acetophenone elution wave. The waves 4 and 5 are the elution waves of the components of the weaning components.

In FIG. 4 is a chromatogram containing lobelin chloride containing flavor and oacharose. Elution wave 3 is acetophenone wave, waves 4 and 5 are elution waves of solvent components.

Preparation of the calibration curve

The following exact concentrations of lobelin chloride in anti-smoking chewing gums were used: 0.03, 0.045, 0.060, 0.075, 0.09 per 100 g of chewing gum. The chewing gums gently and gently weigh and are weighed preone into an Erlenmayer flask with a baffle and baffle plug. 20 ml of 95% alcohol, 10 ml of 1 mol of potassium hydroxide and 10 ml of deotified water are added. The mixture was boiled under reflux for 4 hours. Subsequently, 100 ml of deionized water are added and deodorization is continued after replacement of the bad condenser with the descending condenser. Collect the distillate in a 100 ml volumetric flask up to 100 ml.

From this solution, α is pipetted 10 µl and loaded α on a liquid chromatograph column. The chromatographic separation and determination of acetophenone, which was prepared from lobelin o and makes the following opooob. Uses oa liquid chromatograph with variable wavelength detector. The area of the peaks about and evaluated by the integrator. It will use a x25x25 cm column. 10 µm bead packed with chemically coupled octadecylsilane and a silica gel column. The solvent is: methanol - water 1: 2. Working at 20 ° C at 12.7 MPa. Dispense about 10 µl of samples with a Hamilton micropipette. The elution weaning flow rate suitable for this and analysis is 1.2 ml / min. Detection of α and δ is done in UV light at 249 nm.

While it was not possible to cut the calibration curve from the values obtained after converting lobelin chloride to acetophenone for

228 259.> Lines after extraction from chewing gum, idealized by and evaluating the results of the analysis from the calibration curve, cut to the above) ooob #

Determination of lobelin chloride in anti-smoking chewing gums g of finely-oat chewing gum and weighing preone into an Erlenmayer flask with a stopper and a stopper. Add about 20 ml of alcohol, 10 ml of 1 M potassium hydroxide solution and 10 ml of de-water. The mixture was boiled under reflux for 4 hours. Then, 100 ml of deionized water are added and distillation is continued after the replacement of the bad condenser to a zootic. Collect the distillate oa in a 100 ml graduated flask, make up to the mark with precision. From this solution oa is pipetted 10 µl and applied oa onto the column · Chromatographic separation and determination of acetophenone prepared from lobelin, and carried out by the previously described method in Example 1, by liquid chromatography method.

The evaluation of the liquid chromatograms obtained from a day makes a calibration curve.

Claims (1)

Method for the determination of lobelin chloride in chewing gum based on its conversion to acetophenone, characterized in that the obtained acetophenone is isolated by distillation and determined. at 20 ° C and 12.7 MPa by liquid chromatography using silica gel with chemically bonded octadecylsilane as the stationary phase and methanol / water 1: 2 as solvent.