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›› 2012, Vol. 7 ›› Issue (5): 220-225.

• Regular Articles • 上一篇    下一篇

Determination of total flavonoids content and quantification of rutin in Momordica tuberosa (Roxb) Cogn. fruits by RP-HPLC

Maheshkumar S. Kale*, Kirti S. Laddha   

  1. Medicinal Natural Products Research Laboratory, Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology (ICT), Nathalal Parekh Marg, Matunga (E), Mumbai 400 019, India
  • 收稿日期:2012-03-16 修回日期:2012-05-08 出版日期:2012-10-20 发布日期:2012-10-20

Determination of total flavonoids content and quantification of rutin in Momordica tuberosa (Roxb) Cogn. fruits by RP-HPLC

Maheshkumar S. Kale*, Kirti S. Laddha   

  1. Medicinal Natural Products Research Laboratory, Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology (ICT), Nathalal Parekh Marg, Matunga (E), Mumbai 400 019, India
  • Received:2012-03-16 Revised:2012-05-08 Online:2012-10-20 Published:2012-10-20
  • Contact: Maheshkumar S. Kale

摘要： In present study, determination of the total flavonoid content and quantitative estimation of rutin in methanolic extract of
Momordica tuberosa (Roxb) Cogn. fruits were performed. The spectrophotometric method based on the formation of complexes
with aluminium chloride was applied to determine the total flavonoids content. Presence of rutin was identified using chemical
test and thin layer chromatography (TLC). Quantification of rutin was done by RP-HPLC method using C-18 column (250
mm × 4.6 mm, 5μm) with an isocratic mobile phase, methanol: water: acetic acid (45:54:1 v/v/v) at a flow rate of 1.0 ml/min.
The wavelength of maximum absorption (λmax) selected for quantification of rutin was 257 nm. The method was validated for
accuracy, precision and specificity. Validation revealed that the method is specific, accurate, precise, reliable and reproducible.
The regression data for the calibration plot showed a good linear relationship with r = 0.9994 for rutin. Limit of detection and
limit of quantification were 2.8 and 9.0 μg respectively. Intra and inter-day RSD of retention times and peak areas was less
than 1.9%. A recovery study was carried out to check the accuracy of the method at three different concentrations and average
recovery was between 95.9 and 103.4% for rutin. Total flavonoid content and the amount of rutin present in M. tuberosa fruits
were found to be 0.47% (w/w) and 0.27% (w/w), respectively.

Abstract: In present study, determination of the total flavonoid content and quantitative estimation of rutin in methanolic extract of
Momordica tuberosa (Roxb) Cogn. fruits were performed. The spectrophotometric method based on the formation of complexes
with aluminium chloride was applied to determine the total flavonoids content. Presence of rutin was identified using chemical
test and thin layer chromatography (TLC). Quantification of rutin was done by RP-HPLC method using C-18 column (250
mm × 4.6 mm, 5μm) with an isocratic mobile phase, methanol: water: acetic acid (45:54:1 v/v/v) at a flow rate of 1.0 ml/min.
The wavelength of maximum absorption (λmax) selected for quantification of rutin was 257 nm. The method was validated for
accuracy, precision and specificity. Validation revealed that the method is specific, accurate, precise, reliable and reproducible.
The regression data for the calibration plot showed a good linear relationship with r = 0.9994 for rutin. Limit of detection and
limit of quantification were 2.8 and 9.0 μg respectively. Intra and inter-day RSD of retention times and peak areas was less
than 1.9%. A recovery study was carried out to check the accuracy of the method at three different concentrations and average
recovery was between 95.9 and 103.4% for rutin. Total flavonoid content and the amount of rutin present in M. tuberosa fruits
were found to be 0.47% (w/w) and 0.27% (w/w), respectively.

Key words: M.tuberosa, HPLC, rutin, quantitative estimation, total flavonoid content

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