Determination of diphenhydramine by HPLC method in biological liquids

Abstract

The aim of the study was to develop an algorithm for directed analysis of diphenhydramine in biological extracts from urine and blood using a unified method of HPLC research. Materials and methods. The extraction of diphenhydramine was carried out with chloroform at pH 9.0. The extracts were purified from impurities by a combination of TLC and extraction with hexane. TLC purification and identification of diphenhydramine were carried out under optimal conditions: organic solvents systemschloroform-methanol (90:10); methanol; methanol-25 % solution of ammonium hydroxide (100:1.5) and chromatographic plates-Sorbfil PTLC-AF-A, Sorbfil PTLC-P-B-UV. For the detection of diphenhydramine, the most sensitive location reagents were used-UV light (λ=254 nm) and Dragendorff reagent in the modification of Mounier. HPLC analysis was carried out on a microcolumn liquid chromatograph "Milichrome A-02" in conditions: reversed-phase variant, column with non-polar sorbent Prontosil 120-5 C 18 AQ, 5 μm; mobile phase in the mode of linear gradient-from eluent А (5 % acetonitrile and 95 % buffer solution) to eluent B (100 % acetonitrile) as during 40 min. The flow rate of the mobile phase has been formed 100 μl/min, injection volume-4 μl. Multichannel detection of the substance was carried out using a UV spectrophotometer at 210, 220, 230, 240, 250, 260, 280 and 300 nm; the optimal value of column temperature-37-40 С and pressure of pump-2.8-3.2 MPa. Results and its discussion. Extraction, purification, identification and quantitative determination of diphenhydramine were carried out according to the developed methods. It is established that when isolating diphenhydramine from blood according to the developed methods it is possible to allocate 34.2-38.4 % of substance   5.69 %, 2.04 % RSDx     and from urine-55.8-60.5 % of substance   3.91 %, 1.40 %. RSDx     Conclusions. An algorithm has been developed for directed analysis of diphenhydramine in biological extracts from urine and blood using a unified HPLC method. Statistical processing of the experimental results indicates the reliability and reproducibility of the technique

Green micellar liquid chromatographic method for simultaneous determination of antihistaminic drugs tripelennamine hydrochloride and diphenhydramine in pharmaceutical gel

“…DPH is official in BP 2022 [13] and USP 40 [2]. The literature review of DPH determination depends on spectrophotometry [14,15], chromatography [16][17][18][19][20][21][22][23][24][25][26], and electrophoresis [27][28][29]. There is no reported method for the simultaneous determination of TRP and DPH.…”

“…DPH is official in BP 2022 [13] and USP 40 [2]. The literature review of DPH determination depends on spectrophotometry [14, 15], chromatography [16–26], and electrophoresis [27–29].…”

Design and validation of analytical methods for quantitative determination of active ingredients in extemporal combined medicine in spray form

“…This method is widely used to determine each of the components of the dosage form. Thus, the HPLC method is proposed for the determination of diphenhydramine hydrochloride in monocomponent ready-made medicinal products [3,4] and biological fluids [5], phenylephrine hydrochloride [6], nitrofural and its metabolites [7], in multicomponent medicinal products -for the determination of phenylephrine hydrochloride in a mixture with anti-inflammatory [8] and anti-inflammatory and antihistamine drugs [9,10], in a combination of nitrofurazone with lidocaine hydrochloride [11] and others.…”