SU1027608A1 - Bis( chlorethyl)amine determination method - Google Patents

Abstract

METHOD FOR DETERMINING DERIVATIVES BIS- (/ i-CHLORETHIL; -AMINE by thin-layer chromatography in a mixture of solvents followed by processing the chromatogram obtained with a chemical reagent is distinguished by the fact that, in order to increase the selectivity and sensitivity of the method, n-butyl mixture is used as a mixture of solvents alcohol, acetic acid and water in the ratio (1:

Description

The invention relates to analytical chemistry, in particular to methods for analyzing bis-1p-chloroethyl amine derivatives, and can be used in factory, control and analytical laboratories and pharmacies. There is a method for determining derivatives. bi5- (o-chloroethyl Lamine by treating the sample being analyzed with a solution of 1-isoniconyl-isopropylhydrin at pH 10-11 and 90-100 ° C. Cl. The disadvantage of this method is its relatively low accuracy. The closest in technical essence and the achieved result is Method for determination of bis - (- chloroethyl} -amine derivatives by thin layer chromatography of an analyzed sample in a mixture of solvents containing chloroform, methanol, followed by chromatography with a Dragendorff reagent 2 Due to the fact that the products of its hydrolysis give a similar reaction, and low sensitivity (0.20-0.28 mg / ml). The aim of the invention is to increase the selectivity and h-sensitivity of the method. The goal is achieved according to the method for determining bis derivatives - (l-chloroethyl) -amine by thin-layer chromatography of the analyzed sample in a mixture of solvents n-butyl alcohol, acetic acid and water in a ratio of 1-3: 1: 4-6, with the following processing of the chromatogram 2-methyl-3-hydroxy- 4,5-di- (hydroxymethyl) -pyridine at 99-100 ° C and pH 11-12, as a mixture of p In a solvent, a mixture of n-butyl alcohol, acetic acid and water is used in a ratio of (1-3): 1: (4-6), 2-methyl-3-hydroxy-4, 5-di- (hydroxymethyl j- pyridine and treatment are carried out at 99-100 ° C and pH 11-12. Example. Definition of sarco lysine. Reagents and solvents: 5% solution of 2-methyl-3-hydroxy-4,5 di - (. Oxime - pyridine hydrochloride, 20% solution of caustic soda, a mixture of n-butyl alcohol,. Acetic acid, water with 2: 1: 5, acetone, 1% solutions of sarcolysine, chlorbutin, dopassali, cyclophosphamide in acetone. 0.5 m 1 is applied to the starting line of chromatographic paper 8 cm in diameter at 23 cm from one another % solution of the specimens under study. The high p1 is immersed and the lower end of the flagellum passing through the center of the circular chromatogram is immersed in the mobile phase. After 10-15 min. take out the chamber, mark the position of the solvent and dry. Dry the chromatograms in UV light and mark the positions of the spots, then show by successively immersing in a 5% solution of 2-methyl-3-hydroxy-4, 5-di (hydroxymethyl) -pyridine hydrochloride and 20% sodium hydroxide solution for 20-30 seconds. The chromatograms are dried in a drying cabinet at 100 ° C and the position of the spot is noted. The chromatogram shows one spot. . The eluate has an absorption band for them, which corresponds to gluconic aldehyde. Measured on an SF-4L spectrometer. PRI mme R 2. Determination of hlorbutin. Reagents and solvents: 5% solution of 2-methyl-3-hydroxy-4,5-di- (hydroxymethyl) pyridine hydrochloride, 20% sodium hydroxide solution, a mixture of n-butyl alcohol, acetic acid, water in a ratio of 2 : 1: 5, acetone, 1% solutions of sarcolysin J chlorbutin, dopane, asale, cyclophosphamide in acetone. On, a start line of chromatographic paper with a diameter of 8 cm at a distance of 23 cm from each other is applied with 0.5 ml of a 1% solution of the studied preparations. Dry and immerse the lower end of the flagellum, pass it through the center of the circular chromatogram into the mobile phase. After 10-15 minutes, the chromatogram is removed from the chamber, the position of the solvent is noted and dried. The dried chromatograms are examined in UV light and the position of the spots is noted, and then they are developed by successively immersing in a 5% solution of 2-methyl-3-hydroxy-4,5-di- (hydroxymethyl-pyridine hydrochloride). and a 20% caustic soda solution for 20-30 C. The chromatographic we are dried in a drying oven at and mark the position of the spot. There is a spot on the chromatogram, the presence of which is confirmed by two methods. The eluate has an absorption band of DTS1. nm, which corresponds to glutaconic aldehyde. Measured on an SF-4A optical spectrophotometer with at. nm. EXAMPLE 3 Determination of dopane Reagents and solvent: 5% solution of 2-methyl-3-hydroxy-4,5-di- (hydroxy methyl. pyridine hydrochloride, 20% sodium hydroxide solution, a mixture of n-butyl alcohol, acetic acid, water in a ratio of 2: 1: 5, acetone, 1% solutions of sarcolysin, Chlorobutin, dopane, asale, cyclophosphamide in acetone. On the start line of chromatographic paper with a diameter 8 cm at a distance of 23 cm from the institute, 0.5 ml of a 1% solution of the investigational preparations was applied to one another. The lower end of the flagellum, passing through the center of a circular chromatogram, is dried and immersed in the mobile phase.

After 10-15 minutes, the chromatogram is removed from the chamber, the position of the solvent is noted and dried. The dried chromatograms are viewed under UV light and the positions of the spots are noted, and then developed by successively immersing in 5% solution of 2-methyl-3-hydroxy-4., 5-di- {hydroxymethyl-pyridine hydrochloride and 20% a solution of caustic soda at 2030 s. The chromatograms are dried in a drying cabinet at 100 ° C and the position of the spot is noted.

There is one spot on the chromatogram, the presence of which is confirmed by two methods. The eluate has an absorption band of nm that

It is responsible for glutaconic aldehyde. The optical density at A 360 nm is measured on an SF-4A spectrophotometer.

PRI me R 4. Definition asale

Reagents and solvents: 5% solution of 2-methyl-3-hydroxy-4,5-di- (hydroxy;;) -pyridine hydrochloride, caustic soda solution, a mixture of n-butyl alcohol, acetic acid, water in a ratio of 2: 1: 5, acetone, 1% solutions of sarkolizin, chlorbutin, dopane, asale, cyclophosphamide in acetone.

On the start line of chromatographic paper with a diameter of 8 cm, at a distance of 23 cm, 0.5 ml of a 1% solution of the studied preparations is applied one to another. They are dried and the lower end of the flagellum passing through the center of the circular chromatogram is immersed in the mobile phase.

. After 10-15 minutes, the chromatogram is removed from the chamber, the position of the solvent is noted and dried. The dried chromatograms are scanned in the UV light and the positions of the spots are noted, and then developed by successive immersion in a 5% solution of 2-methyl-3-hydroxy-4,5-di- (XmethylJ-pyridine hydrochloride and 20% caustic soda solution for 20-30 s. Chromatograms are dried in an oven at 100 ° C and the position of the spot is noted.

There is one spot on the chromatogram which is confirmed by two methods. The eluate has a half absorption of them, which corresponds to

glutaconic aldehyde. Optical density nm is measured on an SF-4A spectrophotometer.

Example sT Definition of cyclophosphamide.

Reagents and solvents 5% solution of -2-methyl-3-windows-4,5-di- (hydroxy methyl J-pyridine hydrochloride, 20% sodium hydroxide solution, a mixture of n-butyl alcohol, acetic acid, water in a ratio of 2: 1: 5, acetone, 1% solutions of sarcolysin, chlorbutin, dopane, asale, cyclophosphamide in acetone.

Chromatographic paper with a diameter of 8 cm at a height of 2-3 cm from one another is applied to the start line with 0.5 m of a 1% solution of the studied preparations. The lower end of the flagellum, which passes through the center of the circular xpoMaTorpaNw.Kj, is dried and immersed in the mobile phase.

After 10-15 minutes, the chromatogram is removed from the chamber, the position of the solvent is noted and dried. The dried chromatograms are viewed under UV light and the positions of the spots are noted, and then they are developed by successively immersion in a 5% solution of 2-methyl-3-hydroxy-4,5-di- (hydroxymethyl) -pyridine hydrochloride and 20% caustic soda solution for 2030 s. The chromatograms are dried in a drying oven at and mark the position of the spot.

There is one spot on the chromatogram, the presence of which is confirmed by two methods. The eluate has an absorption band with 360 im, which corresponds to glutaconic aldehyde. The optical density at the SF-4A spectrophotometer is measured at

Vax 360 nm.

The results of the spectrophotometric determination of alkylating agents are given in Table. one.

These sensitivity reactions are shown in Table. 2

Comparative data of the quantitative content of anticancer drugs and products from hydrolysis when choosing the optimal ratio of solvents are given in Table 3.

The use of the proposed method for the analysis of anticancer drugs in comparison with the known method has greater selectivity and sensitivity. 51027608 1.. The preparation Sarkolizin Chlorbutin Dopan Asaley Cyclophosphamide The preparation Sensitizer known Sarcolizin0.20 Chlorobutyl0.18 Dopant0.17 Asaley0.28 Cyclophosphamide 0.26 Quantitative preparation ° ° - "° -1t 7e C1s SopSeactive production of substances hydrolysis

1: 1: 6

91.7

1: 1: 6 90.9

1: 1: 6 92.7

1: 1: 6 93.2

1: 1: 6 90.5

16.4 + 7.3% 98.7

2: 1: 5 97.9 2: 1: 5 99.1

2: 1: 5 99.0 2: 1: 5

80,52,734,517,8

90.15,339,421.8 91,43,840,218,5 92 / 15,242,415,1 91,46,245,311,4

7.8 + 8.2% 49.5 + 10.6%

91.2.7,743,954,2 90,08,253,746,1 88,811,059,340,2 86,212,260,639,1, 6 TABLE 1 Table of contents,% 98.73 97.97 99.13 99.02 98.15 T a l and c a 2 ness, according to the method , mg / mi proposed 0.033 0.040 0.010 0.084 0.025 Table 3 content with expired ORO “see the validity of the active production of the active production of the guideline

Continued, table. 3

Cyclophosphamide 2: 1: 5

98.1

91.2

53.5

43.2

7.5

Claims (1)

A METHOD FOR DETERMINING BIS- ((b-CHLOROETHYL J-AMINE DERIVATIVES by thin-layer chromatography in a solvent mixture followed by treatment of the obtained chromatogram with a chemical reagent) is distinguished by the fact that, in order to increase the selectivity and sensitivity of the method, a solvent mixture is used a mixture of n-butyl alcohol, acetic acid and water in the ratio (1-3): 1: (4-6), 2'-methyl-3-hydroxy-4,5-di- (hydroxymethyl) - pyridine and processing are carried out at 99-100 ° C and pH 11-12.