Page 37 - _NIPER-G Annual Report 2019-201

Research Area-3

Analytical Method Development and Validation of Reverse-Phase High-Performance
Liquid Chromatography (RP-HPLC) Method for Simultaneous Quantifications of
Quercetin and Piperine in Dual-Drug Loaded Nanostructured Lipid Carriers
Quercetin and piperine are often used as an add-on therapy for various diseases, however
both drug exhibits poor aqueous solubility and photosensitivity issue. Therefore, the aim of the
present study is to improve the pharmaceutical challenges by incorporating both the drugs in
nanostructured lipid carriers (NLCs) and to develop a sensitive, selective, accurate and
precise reverse-phase high performance liquid chromatography (RP-HPLC) method for the
simultaneous analysis of both drugs in NLCs. Effective chromatographic separation of
quercetin and piperine was achieved on Hypersil gold C-18 column and mobile phase
consisting of a mixture of acetonitrile and HPLC grade water (pH 2.6, adjusted with 2%v/v
glacial acetic acid) in an isocratic elution mode. The flow rate of the mobile phase was 1
°
mL/min, column temperature at 35±0.2 C and the injection volume was 20 µL. The retention
time for quercetin and piperine were found to be at 2.80 min and 10.36 min, respectively and
detected at an isobestic wavelength of 346 nm using a photodiode array (PDA) detector. The
method was found to be specific for the simultaneous analysis of quercetin and piperine in
presence of NLCs matrix, accurate (>90%) and precise (%RSD <2%). The validated RP-HPLC
method effectively utilised to determine the percentage drug entrapment efficiency cum
percentage drug loading of quercetin and piperine in NLCs enriched formulations along with
the secondary estimation of in vitro cumulative percentage drug release study. The results
were found to be reliable, hence the validated RP-HPLC method could be further used for the
simultaneous detection and quantification of both these drugs in other lipid-based nano-
formulations.

Project Funding Acknowledgement:
National Mission on Himalayan Studies [File No: GBPI/NMHS-2017-18/HSF-02], Ministry of
Environment, Forest and Climate Change, Govt. of India.

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