Simultaneous estimation of (α + β) boswellic acids, β-asarone, isoeugenol, 6-gingerol, and piperine in Majun Nisyan by high-performance thin-layer chromatography

Abstract

Majun Nisyan (MJN) is a polyherbal formulation used in Unani medicine for its therapeutic efficacy against Nisyan (forgetfulness/dementia). In the current work, an effort was made to develop and validate a new high-performance thin-layer chromatography (HPTLC) method for the quantitative estimation of the marker compounds (α + β) boswellic acids, β-asarone, isoeugenol, 6-gingerol and piperine from the formulation of MJN. The method employed HPTLC plates precoated with silica gel 60 F₂₅₄ as the stationary phase. The solvent system comprised of toluene‒ethyl acetate‒chloroform‒acetic acid (8:2:5:0.1, V/V). This method yields compact spots for (α + β) boswellic acids (R_F 0.102 ± 0.01), β-asarone (R_F 0.982 ± 0.04), isoeugenol (R_F 0.850 ± 0.03), 6-gingerol (R_F 0.698 ± 0.03) and piperine (R_F 0.355 ± 0.01). A CAMAG TLC Scanner 4 was used to carry out the densitometric scanning at 254 nm. The International Council for Harmonisation’s Q2(R1) guideline was used to validate the HPTLC method. Each gram of the MJN formulation was found to contain 0.0248 ± 0.0053 mg (α + β) boswellic acids, 0.0520 ± 0.0069 mg β-asarone, 0.0487 ± 0.0041 mg isoeugenol, 0.0468 ± 0.0023 mg 6-gingerol and 0.0609 ± 0.0084 mg piperine. The developed and validated HPTLC method was found to be simple, inexpensive, sensitive, precise and accurate. This method can be easily applied to routine control analysis of all commercially available Ayurvedic and herbal formulations containing (α + β) boswellic acids, β-asarone, isoeugenol, 6-gingerol and piperine.

Graphical abstract