Rapid HPLC Method for Determination of Vitamin C, Phenolic Acids, Hydroxycinnamic Acid, and Flavonoids in Seasonal Samples of Emblica officinalis Juice

Abstract

The work proposes a simple method for simultaneous detection and quantification of vitamin C (ascorbic acid), phenolic acids (gallic acid and ellagic acid), hydroxycinnamic acid (chlorogenic acid), and flavonoids (myricetin, quercetin, and kaempferol) in seasonal samples of emblica juice. The compounds were separated by an intangible curved gradient of 0.1% orthophosphoric acid in water (v/v) and acetonitrile as mobile phase A and B using Zorbax SB RP C-18 column at a wavelength of 254 nm in 18 min. The assay was optimized by varying the mobile phase, gradient type, and detection wavelength. The method was validated in terms of linearity, precision, detection limits, and quantification limits. Good linear response was observed over the range specified for all the analytes, as confirmed by the correlation coefficient which ranged from 0.991 and 0.995. The limit of detection and limit of quantification were found to be in the range of 0.129–0.685 µg mL⁻¹ and 0.43–2.883 µg mL⁻¹, respectively. Complete resolution of the phenolic compounds was achieved without the need of pre-analysis processes. The overall precision values obtained for standards and samples were within the range of 0.01–0.2 and 0.15–0.23, respectively.

Keywords:

• curved gradient
• Emblica officinalis juice
• HPLC
• method development
• total polyphenol content
• vitamin C

Acknowledgments

Authors are thankful to the director, Central Scientific Instruments Organisation (CSIO), for infrastructure facilities.

Notes

*S1 mean value (2011) stock ± SD, where n = 9.

**S2 mean value (2012) stock ± SD, where n = 9.

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