Volume No. :   8

Issue No. :  3

Year :  2018

ISSN Print :  2231-5667

ISSN Online :  2231-5675


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Development of a New Isolation Technique and Validated RP-HPLC method for Quercetin and Kaempferol from Azadirachta indica leaves



Address:   Om Prakash1*, Debarshi Kar Mahapatra2, Ruchi Singh1, Namrata Singh1, Neelam Verma1 Akash Ved1
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
DOI No: 10.5958/2231-5675.2018.00030.3

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 ?max of 259 nm and 364 nm and the isolated quercetin displayed ?max of 256 nm and 365 nm, respectively. The standard kaempferol exhibited ?max of 267 nm and 368 nm whereas the isolated kaempferol presented ?max 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.
KEYWORDS:
Quercetin, Kaempferol, Azadirachta indica, RP-HPLC, Flavonoid, Phenolic.
Cite:
Om Prakash, Debarshi Kar Mahapatra, Ruchi Singh, Namrata Singh, Neelam Verma Akash Ved. Development of a New Isolation Technique and Validated RP-HPLC method for Quercetin and Kaempferol from Azadirachta indica leaves. Asian J. Pharm. Ana. 2018; 8(3): 164-168.
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