A High-Performance Liquid Chromatography-Mass Spectrometry Method Development for the Quantitative Determination of Enalapril Maleate From Caco-2 Cell Monolayers

Abstract

Objective: A simple, rapid high-performance liquid chromatography-mass spectrometry (HPLC MS/MS) method was developed for the determination of enalapril maleate from confluent Caco-2 monolayers and aqueous solution.Methods: Chromatography was achieved on Discovery C18, 50×2.1 mm, 5 μm column. Samples were chromatographed in a gradient mode (eluent A [acetonitrile-water-formic acid, 5: 95:0.1 v/v] and eluent B [acetonitrile-formic acid, 100:0.1 v/v]). The initial content of the eluent B is 0%, which increases linearly by 1.0 min to 100% and 1.01 min returns to the initial 0%. The mobile phase was delivered at a flow rate of 0.4 ml/min into the mass spectrometer ESI chamber. The sample volume was 5 μl.Results: Under these conditions, enalapril maleate was eluted at 1.52 min. According to the Caco-2 test results, enalapril showed low permeability. It should be noted that the recovery value for enalapril is 100.62 %. Low in vitro permeability data for enalapril are in good agreement with the literature data.Conclusion: From results of the analysis, it can be concluded that developed method is simple and rapid for determination of enalapril maleate from confluent Caco-2 monolayers and aqueous solution. Acquired results demonstrate that proposed strategy can be effortlessly and advantageously applied for examination of enalapril from Caco-2 cell monolayers.

Development of Methods of Analysis of Biological Active Substances for Standardization of Dense Extract of Creeping Thyme

“…Approaching the standardization of liquid and dense thyme creeping extract, we considered the possibility of identifying the same BAS, continuing the proposed approach in the HMP chain extracts [9]. The determination of the amount of flavonoids was carried out by the method of differential spectrophotometry on the spectrophotometer of the brand "Cary-50" [12]. The quantitative content of flavonoids in the dense extract was determined by the reaction of the formation of the colored compound directly of flavonoids with aluminum chloride.…”

“…With a vacuum dilution of 0.8 kp/cm 2 , the gain is about 15-20% in comparison with the vacuum of 0.6 kp/cm 2 Identification and determination of the quantitative content of the analyzed BAS in HMP and their respective extracts were carried out using modern sensitive and selective methods of analysis. The qualitative composition of flavonoids was determined by the thinlayer chromatography method [11,12] in the solvent system ethyl acetate Pformic acid Pwater P (90:6:9). The determined volumes of solutions were applied to chromatographic plates Silica gel F254 of company "Merck" using a device for automatic sampling on "CAMAG Linomat 5" plate, chromatography was carried out in a "GAMAG" chromatography chamber.…”

“…The determination of the amount of flavonoids was carried out by the method of differential spectrophotometry on the spectrophotometer of the brand "Cary-50" [12].…”

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“…Both evaluated drugs were "rapidly dissolving" [ Table 1] because the active pharmaceutical ingredient release at time point 30 min was >85%. [21][22][23][24][25] Table 1.…”

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“…For a successful study of this substance by these methods, usually, derivatization of active groups with one or more reagents is carried out. [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21] However, as practice shows, in case of using tianeptine for the purpose of obtaining a narcotic effect, urine contains products of its metabolism or disintegration.…”