Mamina О.О.

The choice of optimal conditions for identification of chloropyramine hydrochloride by GLC / MS method, suitable

for chemical and toxicological analysis

Ukrainian National University of Pharmacy, t. Kharkov

Given the complexity of the analysis of biological objects for the presence of toxic substances and their metabolites (possibility of falsification tests, small volume, the presence in the sample a significant amount of background of
endogenic and exogenic substances) for reliable results it is important to compare the results of preliminary and confirmatory methods using specific characteristics - mass spectra - retention parameters obtained in the performance chromatography and conduct of computer-based search software libraries).

According to the literature combination of extraction methods and methods gas-liquid chromatography with mass spectrometric detection (GLC/ MS) [1-3] provides reliable and reproducible results relatively the presence of toxic substances in biological objects.

Analysis of chloropyramine hydrochloride GLC/ MS -method performed on a gas chromatograph Hewlett Packard HP-6890 / 5972A with mass selective detector on the basis Kharkov Military Ecology Research Center.

Separation of mixture components performed on capillary column Rxi-5MS (5% Diphenyl Methyl Siloxane), nominal length and diameter - 30 m x 250 µm, nominal film thickness phase – 0,25µm. The standard solutions of chloropyramine hydrochloride in ethanol with a concentration 100,0-200,0 µg / ml were used for chromatography.

As a result of studies found optimal parameters GLC/MS - analysis chloropyramine: parameters of mass selective detector (temperature of ion source and interface lines - 280°С, for detecting ions - scan mass numbers range - 45-450); parameters of gas chromatograph (carrier gas – helium, gas flow rate of carrier - 1 cm³ / min, the temperature of the evaporator - 280°С, volume of sample introduction - 1 µl, chromatography mode - without flow separation); temperature mode (initial temperature - 45°С, time of chromatography at the initial temperature - 2 min, linear increase in temperature - 10°C/min, the final temperature of - 280 °С, time of chromatography at the final temperature - 5 min).

As a result of the identification chloropyramine hydrochloride GLC - method in developed conditions (Fig. 1) is set: retention time 22,82 ± 0,02 min, the number of theoretical plates - 1441,4, coefficient symmetry - 1,0. The detection limit was 0,1 µg / ml, which corresponds to the content in the sample chloropyramine hydrochloride 0,1 ng.

According to the chromatogram conducted searches using libraries of mass spectra "Nist 02 Spectra Lib" and “Wiley 138”.

Retention time, min

Fig. 1 GLC- chromatogram of chloropyramine ethanol solution with a

concentration of 100,0 µg / ml

Established that the mass spectrum chloropyramine in a standard solution for the retention time 22,814 min has a set of specific ion m / z: 58, 71, 79, 89, 99, 125, 219 (Fig. 2), which corresponds to the literature [1].

m/z

m/z

Fig. 2 Mass-spectrum of chloropyramine retention time 22,814 min

Developed conditions of GLC / MS method is recommended to apply after extraction and TLC-treatment of extracts from biological material in parallel with highly sensitive methods HPLC, UV spectrophotometry.

Conclusions:

The choice of optimal conditions for identification of chloropyramine hydrochloride by GLC / MS determines its high sensitivity (0,1 ng in the sample), selectivity, providing the opportunity to get objective results of chemical-toxicological research.

References:

1. Clarke, E.J.C. Isolation and Identification of Drugs in Pharmaceuticals, Body Fluids and Postmortem Material. – London: The Pharm. Press, 2011. – 2463 p.

2. The detection and quantification of ethyl glucuronide in placental tissue and placental perfusate by headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry / J. N. Matlow, K. Aleksa, A. Lubetsky, G. Koren // Canad. J. Clin. Pharmacology. – 2012. - Vol. 19, № 3. - P. 473–482.

3. Determination of seven selected antipsychotic drugs in human plasma using microextraction in packed sorbent and gas chromatography-tandem mass spectrometry / B. M. da Fonseca, I. E. D. Moreno, M. Barroso, S. Costa, J. A. Queiroz, E. Gallar-do //Analyt.and Bioanalyt. Chem. - 2013. - Vol. 405, № 12. - P. 3953–3963.