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Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (9): 1022-1030.doi: 10.3736/jcim20110914

• Original Experimental Research • Previous Articles     Next Articles

Simultaneous determination of quercetin, rutin and kaempferol in the leaf extracts of Moringa oleifera Lam. and Raphinus sativus Linn. by liquid chromatography-tandem mass spectrometry

Venkatapura C. Devaraj1(), Burdipad G. Krishna2, Gollapalle L. Viswanatha3#br#   

  1. 1. Department of Pharmacology, Bioneeds Laboratory Animals and Preclinical Services, Bangalore 562111, India
    2. Department of Pharmacognosy, R. R College of Pharmacy, Bangalore 560090, India
    3. Department of Pharmacology, Peopled Education Society College of Pharmacy, Bangalore 560050, India
  • Received:2011-04-23 Accepted:2011-06-26 Online:2011-09-20 Published:2011-09-15
  • Contact: C. Devaraj Venkatapura

Objective: To develop a rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to analyze quercetin (QU), rutin (RU) and kaempferol (KA) simultaneously in the leaf extracts of Moringa oleifera Lam. and Raphinus sativus Linn. 
Methods: Samples were prepared by extracting the leaves of the M. oleifera and R. sativus by cold-maceration technique using 90% ethanol. Chromatographic separation was operated with a mixture of 0.2% formic acid in water and acetonitrile at a flow rate of 0.4 mL/min on a Phenomenex Gemini C18 column with a total run time of 5.01 min. 
Results: The MS/MS ion transitions monitored were 303.03 to 153.1 for QU, 611.1 to 303.1 for RU, 287.1 to 153.2 for KA and 180.1 to 110.1 for internal standard. The lower limit of quantitation achieved for QU, RU and KA was 5 ng/mL and the linearity was observed from 5 to 2 000 ng/mL. The correlation coefficients of linear regression analysis were 0.9946, 0.9951 and 0.9969 for QU, RU and KA, respectively. 
Conclusion: The results indicate that the LC-MS/MS method is fast and sensitive and may provide excellent specificity for simultaneous determination of QU, RU and KA in leaf extracts of M. oleifera and R. sativus.

Key words: Moringa oleifera, Raphinus sativus, plant extracts, quercetin, rutin, kaempferol, chromatography, liquid, tandem mass spectrometry

Figure 1

Structures of quercetin, rutin and kaempferol"

Figure 2

Typical multiple reaction monitoring chromatograms of quercetin (left panel) and internal standard (right panel) in a) mobile phase, b) reference standard, c) Raphinus sativus leaf extract, and d) Moringa oleifera leaf extract"

Figure 3

Typical multiple reaction monitoring chromatograms of rutin (left panel) and internal standard (right panel) in a) mobile phase, b) reference standard, c) Raphinus sativus leaf extract, and d) Moringa oleifera leaf extract"

Figure 4

Typical multiple reaction monitoring chromatograms of kaempferol (left panel) and internal standard (right panel) in a) mobile phase, b) reference standard, c) Raphinus sativus leaf extract, and d) Moringa oleifera leaf extract"

Figure 5

Representative spectra of product ion of quercetin"

Figure 6

Representative spectra of product ion of rutin"

Figure 7

Representative spectra of product ion of kaempferol"

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