Development and Validation of an RP- UFLC Method for Simultaneous Estimation of Quercetin and Rutin
Hygeia.J.D.Med. 2013; 5(1):113-120
Shanmugam. R1,Gowthamarajan. K 1*, Priyanka D.L1, Madhuri.K2,Hemnath.E3, Dhanabal. SP 4
1.Department of Pharmaceutics, JSSCollege of Pharmacy (JSS University, Mysore, Karnataka), Rocklands, Ooty- 643001, India.
2.Department ofPharmacology, JSS College of Pharmacy (JSS University, Mysore, Karnataka), Rocklands, Ooty 643001, India.
3. Department of Pharmaceuticalchemistry, JSS College of Pharmacy (JSS University, Mysore, Karnataka), Rocklands,Ooty 643001, India.
4.Department of Phytopharmacy & Phytomedicine, JSSCollege of Pharmacy (JSS University, Mysore, Karnataka), Rocklands, Ooty ,India.
Article history: Received: 2 December 2012, revised: 12 January 2013, accepted: 5 February 2013, Available online: 3 April 2013
Plan: Quercetin and rutin are the flavonoides possess good therapeutic properties.The main objective of our work wasto develop a simple, rapid and sensitive UFLC method for simultaneous estimation of quercetin and rutin.
Methodology: A simple chromatographic separation was done by using a reverse phase C8 column (250 x 4.6mm) by using isocratic flow with the constituted a mixture of 25 mM potassium dihydrogen ortho phosphate (pH4) and acetonitrile mobile phase at the ratio of70:30 with the flow rate of l ml/min. Detection was carried out at 230 nm.Different validation parameters such as specificity,sensitivity, linearity, accuracy, precision, ruggedness and robustness were performed.
Outcome: Optimized Chromatographic conditions were achieved and results showed good peakresolution between quercetin and rutin. The retention time was found at 7.4 min and 2.8 min. The LOD for quercetin and rutin was found to be 10 ng and 15 ng& LOQ was found to be 90 ng and 100 ng. The calibration curve was fixed at 0-100000 ng/ml. With change in analytical method parameters, analyst,laboratory conditions did not affect the results. % CV was calculated for all the parameters and found to be within specification limits. The developed method can be used for the quantitative and qualitative estimation of quercetin and rutin for the herbal medicinal products.
Keywords: Herbal medicinal products,ICH, Quercetin, Rutin, UFLC
Shanmugam. R,Gowthamarajan. K , Priyanka D.L, Madhuri.K, , Hemnath.E, Dhanabal.SP. Development and Validation of a RP UFLC Method forSimultaneous Estimation of Quercetin and Rutin .Hygeia.J.D.Med. 2013; 5(1):113-120.Available at http://www.hygeiajournal.com / Article ID- Hygeia.J.D.Med/95/13.
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Researcher ID: J-7381-2012
Typical HPLC chromatogram of Standard Quercetin and Standard Rutin