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  • Determination of saponins and flavonoids in ivy leaf extracts using HPLC-DAD.

Determination of saponins and flavonoids in ivy leaf extracts using HPLC-DAD.

Journal of chromatographic science (2014-07-02)
Miao Yu, Young June Shin, Nanyoung Kim, Guijae Yoo, SeonJu Park, Seung Hyun Kim
ABSTRACT

A new method for the determination of six compounds, chlorogenic acid, rutin, nicotiflorin, hederacoside C, hederasaponin B and α-hederin, in ivy leaf extracts using high-performance liquid chromatography with diode array detector was developed. The chromatographic separation was performed on a YMC Hydrosphere C18 analytical column using a gradient elution of 0.1% phosphoric acid and acetonitrile. The method was validated in terms of specificity, linearity (r(2) > 0.9999), precision [relative standard deviation (RSD) < 0.36%] and accuracy (97.4-103.8%). The limit of detection and limit of quantification were <20.32 and 61.56 ng for all analytes, respectively. The tested compounds were found to be stable in the ivy leaf extract from 0 to 48 h, and the RSD value for each compound was <0.90%. The validated method was successfully applied to quantify all six compounds in a 30% ethanol ivy leaf extract and 13 ivy leaf extract products. The results showed that all the tested products satisfied the minimum requirement for the content of hederacoside C. However, there were some differences between the contents of other constituents.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Phosphoric acid solution, 85 wt. % in H2O, FCC, FG
Sigma-Aldrich
Phosphoric acid solution, suitable for NMR (reference standard), 85% in D2O (99.9 atom % D), NMR tube size 4.2 mm × 8 in. , WGS-5BL Coaxial NMR tube
Sigma-Aldrich
Methanol, anhydrous, 99.8%
Sigma-Aldrich
Chlorogenic acid, ≥95% (titration)
Sigma-Aldrich
Methanol, purification grade, 99.8%
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Phosphoric acid solution, suitable for NMR (reference standard), 85% in D2O (99.9 atom % D), NMR tube size 3 mm × 8 in.
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Phosphoric acid solution, suitable for NMR (reference standard), 85% in D2O (99.9 atom % D), NMR tube size 5 mm × 8 in.
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Phosphoric acid, ≥85 wt. % in H2O, ≥99.999% trace metals basis
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Acetonitrile, anhydrous, 99.8%
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Phosphoric acid, crystalline, ≥99.999% trace metals basis
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Phosphoric acid-16O4 solution, 70 wt. % in D2O, 99.9 atom % 16O
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