Development of a New Isolation Technique and Validated RP-HPLC method for Quercetin and Kaempferol from Azadirachta indica leaves Prakash Om1,*, Mahapatra Debarshi Kar2, Singh Ruchi1, Singh Namrata1, Verma Neelam1, Ved Akash1 1Goel Institute of Pharmacy and Sciences, Faizabad Road, Lucknow, Uttar Pradesh, India 2Department of Pharmaceutical Chemistry, Dadasaheb Balpande College of Pharmacy, Nagpur, 440037, Maharashtra, India *Corresponding Author E-mail: opverma2007@gmail.com
Online published on 24 October, 2018. Abstract The aim of this study was to develop a new method for isolating the two major flavonoids, quercetin and kaempferol by using the RP-HPLC technique from Azadirachta indica leaves. The flavonoids were initially separated and identified by analytical column while isolated by the preparative column. The quercetin and kaempferol were identified by an external standard analytical RP-HPLC procedure. The fraction collection was set up through software and the peaks collected in the chromatogram were 7.42 min for quercetin and 11.39 min for kaempferol. The purity of the collected compounds showed UV area % purity of 86.67 and 89.23 for quercetin and kaempferol, respectively. The standard quercetin demonstrated λmax of 259 nm and 364 nm and the isolated quercetin displayed λmax of 256 nm and 365 nm, respectively. The standard kaempferol exhibited λmax of 267 nm and 368 nm whereas the isolated kaempferol presented λmax of 259 nm and 361 nm, respectively. Both the FT-IR spectra (standard and isolated) of quercetin and kaempferol superimpose with each other. The prominent spectral peaks were located at >3300 cm−1 (indicating-OH groups) and 1600–1700 cm−1 (representing the carbonyl function). The method has been found to be precise, accurate, linear, robust, economic, sensitive, and can rapidly isolate both the key phytoconstituents from the extracted natural materials. Thus, the research opens avenues for the application of this method in industry-based functions as well as quality control measures at varied operability scale. Top Keywords Quercetin, Kaempferol, Azadirachta indica, RP-HPLC, Flavonoid, Phenolic. Top |