CS261315B1 - Method for Analysis of Diethyllysergamide Azobenzenaphthalenesulfonates - Google Patents

Abstract

The solution falls into the field of clinical biochemistry, pharmaceutical analysis, chemical toxicology and chemical control. The previously known methods of detection, identification and preparation of lysergic acid derivatives are based on procedures that require qualified personnel, special equipment and are time-consuming. The shortcomings are eliminated by a method that depends on the fact that an aqueous solution of diethyllysergamide is treated with an aqueous or methanol solution of azobenzenenaphthalenesulfonate of the formula 1, where R is methyl, phenyl, carboxyl, hydroxyl, nitro group, amino group, sulfo group, while the number of substituents R is expressed by the number η = 1 to 4 and the formed ionic associate is shaken at pH 1 to 5.5 with benzene, chloroform, 1,1,2,2-tetrachloroethane, xylene or toluene, after which the yellow to red colored solution of the ionic associate of diethyllysergamide with azobenzenenaphthalenesulfonate is measured colorimetrically. The procedure also brings the benefit of the possibility of express evaluation of samples containing diethyllysergamide, from a limiting concentration of 1.10-6 mol.dm-3 with an accuracy of 1 5 I, expressed by relative standard deviation. It meets the requirements of automation.

Description

The present invention relates to a method for the analysis of diethyllysergamide azobenzenaphthalenesulfonates of the general formula ^{R 11 -C} 6 ^H 4 ^{N = N} - ^C 8 ^H 4 ^{R 11}

The radical R is methyl, phenyl, carboxyl, hydroxyl, nitro, amino, sulfo. The number of substituents is expressed by the number η = 1 to 4.

The invention provides for the simple expression, sensitive spectrophotometric determination and detection of a group of biologically active compounds that find use in medicine, pharmacy, can also be misused as drugs and means in diverse events or for terrorist attacks. Thus, the invention may be included in the fields of clinical biochemistry, pharmaceutical analysis, chemical toxicology, chemical control of armed forces and civil defense services.

Hitherto known methods for the detection, identification and determination of lysergic acid derivatives, including lysergamides, are based on classical color reactions with sulfuric acid and formaldehyde [WEBER, Q., W., A. - HEVERAU, J., E .: J. Pharm. Sci 62, 1973, p. 1 174], modifications according to the Winkler reaction [FRIEDMAN, M. - Finley JW, J. Agr. Food Chem. 19, 1971, p. 62β], Keller reactions [cONCON J. M .: Anal. Biochem. 67, 1975, pp. 206, FAUBERT M.: J.

Pharm. Pharmacol. 26, 1974, pp. 637j.

Other physicochemical methods of identification, detection or determination have been developed for the analysis of LSD and other lysergic acid derivatives. Thin-layer and column chromatography including mass spectrometry is suitable and reliable. Less suitable is gas chromatography [sPEHLING A., J.: J. Chromatogr. Sci. 10, 1972, p. Volumetric methods are suitable for semi-quantitative methods of analysis. Is it necessary to work with a solution volume of at least 5 µ? and a concentration of -4 -3 above 5.10 mol.dm. A suitable method for potentiometric titration of generally organic bases with sodium tetrafenylborate to a potassium ion selective electrode has been published [VYTŘÁS K .: Coll. Czech. Chem. Commun. 42, 1977, p. Spectrofluorimetry methods have also been developed for the analysis of lysergamides [BAUDOT Ph. - ANDRÉ.J. C .: Analyt. Letters A 15, 1982, pp. 471, infrared spectrometry [CROMC. C. TURNEYF. G .: J. Forensic. Sci. 15, 1970, p. 530, mass spectrometry, BELXMAN S. W., J. Assoc. Offic. Analyt. Chem. 51, 1968,

164.

J

These methods of analysis are time-consuming, requiring qualified staff and special analytical equipment. This does not give the possibility to use it in the laboratory operation at the basic stage, where only the thin-layer chromatography is available from the mentioned methods. The above drawbacks are overcome by the analysis method of the invention.

The present invention is a method of analyzing diethyllysergamidu azobenzennaftalensulfonany which consists in that the aqueous solution diethyllysergamidu treated with an aqueous or methanolic solution azobenzennaftalensulfonanu formula R ^ _n -CgH -N = N-CGH ^ _n -R wherein R is methyl, phenyl, carboxyl , hydroxyl, nitro, amino, sulfo, wherein the number of substituents R is expressed as η = 1 to 4 and the resulting ionic associate is shaken at pH 1 to 5.5 with benzene, chloroform, 1,1,2,2-tetrachloroethane, carbon tetrachloride, xylene or toluene, followed by a yellow to red solution of the diethyllysergamide ionic associate with azobenzene naphthalenesulphonate is measured colorimetrically.

The advantage of the elaborated procedure is the possibility to determine diethyllysergamide up to • 6 - 3 concentration

1.10 ⁰ mol.dm. The assay does not interfere with the presence of codeine, physostigmine, morphine, yohimbine,

-3-3 dihydroergotamine, atropine, hyoscyamine, caffeine, nicotine up to a concentration of 5.10 mol.dm in the analyzed process solution. The time required to perform the analysis is 4 minutes. Using the method of analysis according to the invention has the advantage of expressing evaluation of blood and urine samples as process solutions containing diethyllysergamide up to a limit concentration of 6 -3 +, 5.10 mol.dm, with a precision of the method expressed as a relative standard deviation of - 5%.

Example 1-3 -3

A urine solution containing 1.10 mol.dm diethyllysergamide in the sample was prepared.

3

A 0.2 cm sample solution was pipetted and 1.6 cm of citrate buffer at pH2.2, 0.2 cm ^{3 of an} aqueous solution of 3- (5'-chloro-2'-hydroxynaphthalene) -5,5-hydroxy-naphthalene was added. Sodium 2,7-diq sulfonate and shaken with 2 cm of chloroform for 1 minute. After removal of the aqueous layer, absorbance at 580 nm was measured. The equation of the calibration line was y »13,000x + + 0.001. The minimum detectable concentration of diethyllysergamide was 5.10 mol.dm. Example 2 *

The urine solution containing 1.10 ^-5 mol.dm ^-3 was pipetted after 0.4 cm ³ and 0.1 cm ^{3 of an} aqueous solution of disodium diphenyl-bis- (1-amino-2-naphthylazo-4-sulfonate) was added and make up to 2,0 cm with buffer solution pH 2,5. The aqueous mixtures were shaken with 2.0 cm ^{3 of} chloroform in one minute. Absorbance of chloroform extracts was measured at 525 nm. The lowest detectable concentration was 4.10 ® mol.dm ³ and the calibration line equation was y = 11 500x + 0.05.

Example 3

-5 —3

0.1 cm of blood plasma containing 1.10 mol.dm diethyllysergamide was pipetted and 0.1 cm ^{3 of an} aqueous solution of 4.10 ^-4 mol.dm ^-3 m-anilinazobenzenesulfonate sodium was added and supplemented with citrate buffer pH 5.0 to 2, 0 cm ³ . The aqueous layers were extracted with 2.0 cm ^{3 of} chloroform, 1,1,2,2-tetrachloroethane, carbon tetrachloride, benzene, toluene and xylene. Extraction into chloroform and 1,1,2,2-tetrachloroethane was performed in 80% yield, carbon tetrachloride 64%, benzene 48%, toluene 27%, xylene 11%. The lowest detectable concentration in the extract was 2.10 ® mol.dm ³ . The equation of the calibration line was y = 15 200x + 0.01.

Example 4 ’-5 —3

0.4 cm of blood plasma containing 5.10 mol.dm diethyllysergamide 3 was pipetted off and 0.1 cm @ 3 of an aqueous solution of dimethylanilinazobenzene added and supplemented with citrate buffer 3, pH 3.0. The aqueous layers were shaken for one minute with 2.0 cm chloroform. After aspiration of the aqueous layer, organic associate solutions were measured at 405 nm. The limit of determination - 6 - 3 '3 was 1.10 mol.dm or 1,2yug.cm. The equation of the calibration line was y = 18,000x + 0.005.

Example 5

A sample of diethyllysergamide was imitated so that 1.0 g of sucrose was homogenized with

100 mg diethyllysergamidium tartrate. A 10 mg sample was weighed, dissolved in water and replenished

3 3 per 100,0 cm. For the determination, 0.1 cm was pipetted, 0.1 cm methanol solution -3-3 was added

4.10 mol.dm sodium p- (2-hydroxy-1-naphthylazo) benzenesulfonate and supplemented with citrate buffer to 2,0 cm ³ . Chloroform extracts were measured at 400 nm. The relative standard deviation was 5.09%. The equation of the calibration line was y = 9 800x + 0,001.

The method of analysis according to the invention satisfies both automation and efficiency requirements by its methodical arrangement. The procedure is modified so that it can be secured by domestic chemicals and is applicable to spectrophotometers, photometers and colorimeters with which the basic operational toxicology and biochemical laboratories are equipped. The method is suitable for complementing a set of methods of analysis in pharmacy, forensic medicine, clinical biochemistry, chemical toxicology, chemical control, by armed forces or civil defense services. The sensitivity which is ensured according to the method of analysis allows the method according to the invention to be used up to a -6-3 limit concentration of 1.10 mol.dm diethyllysergamide in a process sample of blood plasma, urine or other body fluid.

Claims (1)

Process analysis diethyllysergamidu azobenzennaftalensulfonany characterized ie the aqueous solution diethyllysergamidu treated with an aqueous or methanolic solution azobenzennaftalensulfonanu formula R _n -CgHj-N "N-Cgll4 _n-R wherein R is methyl, phenyl, carboxyl, hydroxyl, nitroskuplna, amino, sulfoekupina, wherein the number of substituents R is expressed as η = 1 to 4 and the resulting ionic associate is shaken out at pH 1 to 5.5 with benzene, chloroform, 1,1,2,2-tetrachloroethane, tetrachloroethane, xylene or toluene, followed by yellow to red the colored solution of the ionic association of diethyllysergamide with azobenzenaphthalenesulfonate is measured colorimetrically.