Simultaneous determination of six active components in traditional herbal medicine 'Oyaksungisan' by HPLC-DAD
ABSTRACT In this study, an effective high performance liquid chromatography-diode array detector (HPLC-DAD) method was established for simultaneous determination of six marker compounds, ephedrine hydrochloride, 6-gingerol, glycyrrhizin, hesperidin, imperatorin and ferulic acid, in a Korean traditional prescription, Oyaksungisan, which is used for hemiplegia, arthralgia and paralysis. The six marker compounds of Oyaksungisan were separated on a LUNA C18 column (S-5 μm, 4.6 mm I.D. × 250 mm) at a column temperature of 35°C. The gradient elution was composed of water with 0.1% trifluoroacetic acid and methanol. The detection UV wavelengths were set at 207, 250, 280 and 320 nm. Calibration curves for the six compounds showed good linear regressions (R (2) > 0.9999). The limits of detection and limits of quantification were within the ranges 0.003-0.01 and 0.01-0.04 μg/ml, respectively. The relative standard deviation (RSD) values of intra- and inter-day testing were within the ranges 0.10-1.82 and 0.04-1.59%, respectively. The results of the recovery test were 95.05-104.27% with a RSD value of 0.11-1.85%. In conclusion, the simultaneous determination method developed was useful in the quality evaluation of Oyaksungisan.
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ABSTRACT: Background: Jaeumganghwa-tang (JEGH) is a traditional Korean herbal medicine for the treatment of chronic bronchitis, nephritis and diabetes mellitus. Objective: A high performance liquid chromatography-diode array detector (HPLC-DAD) method was developed for simultaneous determination of 11 major compounds such as 5- hydroxymethylfurfural, mangiferin, paeoniflorin, nodakenin, naringin, hesperidin, decursinol, berberine, glycyrrhizin, atractylenolide III and decursin, in JEGH. Materials and Methods: The separation was conducted on Shishedo C 18 column with gradient elution of 0.1% trifluoroacetic acid-acetonitrile. Detection of wavelength was set at 205, 250, 280 and 330 nm. Results: The developed analysis showed a good linearity (R-2 > 0.9997). The range of limit of detection and limit of quantification were observed from 0.04 to 0.43 and from 0.11 to 1.30, respectively. The intra- and inter-day test relative standard deviations (RSD) were less than 3% and the accuracy was 95.98-108.44%. The recoveries were between 92.75% and 109.19% and RSD range of recoveries was measured from 0.52% to 2.78%. Conclusion: This HPLC-DAD method can be successfully applied for simultaneous determination of 11 major compounds in JEGH samples.Pharmacognosy Magazine 04/2014; 10(Suppl 2):S256-63. DOI:10.4103/0973-1296.133267 · 1.11 Impact Factor
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ABSTRACT: The aim of this study was to determine quantitatively the 12 marker compounds in Palmijihwang-hwan (PJH) using a high performance liquid chromatography-photodiode array detector. The separation of marker compounds was performed on a reversed-phase C18 column for gallic acid, 5- hydroxymethylfurfural (5-HMF), morroniside, loganin, paeoniflorin, mesaconitine, benzoic acid, coumarin, cinnamic acid, cinnamaldehyde, and paeonol, and an amino (NH2) column for allantoin. The correlation coefficient of marker compounds was $0.9993, which means good linearity. The limit of detection and limit of quantification values were in the ranges from 0.01–0.25 mg mL�1 and 0.02–0.78 mg mL�1, respectively. The within-day precision was 0.06–2.95% and the between-day precision was 0.10–4.44% over five consecutive days. The recovery of marker compounds ranged from 97.20 to 106.36%, with RSD values <3.5%. The repeatability was <2.1% of the RSD value. The quantification results indicated that the quantities of the 12 marker compounds differed between a water extract and commercial granules of PJH. Cinnamaldehyde and paeonol were particularly difficult to determine in commercial granules because of its low LOQ. The evaluation of the Pearson coefficient and principal component analysis showed a clear discrimination between a PJH water extract and commercial granules of PJH. The analytical method established was precise, accurate, and reproducible for evaluating the quality of PJH.Analytical methods 03/2014; 6(11):3763–3771. DOI:10.1039/c4ay00249k · 1.94 Impact Factor
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ABSTRACT: A simple and rapid ultra-high performance liquid chromatographic (UHPLC) method coupled with an ultraviolet detector (UV) has been developed and validated for the separation and determination of 14 major flavonoids ((±)-catechin, (-)-epicatechin, glycitin, (-)-epicatechin gallate, rutin, quercitrin, hesperidine, neohesperidine, daidzein, glycitein, quercetin, genistein, hesperetin, and biochanin A) in herbal dietary supplements. The flavonoids have been separated on a Chromolith Fast Gradient Monolithic RP-18e column utilizing a mobile phase composed of 0.05% trifluoroacetic acid in water and acetonitrile in gradient elution mode. Under these conditions, flavonoids were separated in a 5min run. The selectivity of the developed UHPLC-UV method was confirmed by comparison with ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis. The validation parameters such as linearity, sensitivity, precision, and accuracy were found to be highly satisfactory. The optimized method was applied to determination of flavonoids in different dietary supplements. Additionally, the developed HPLC-UV method combined with 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) assay was used in the evaluation of antioxidant activity of the selected flavonoids. Copyright © 2014 Elsevier B.V. All rights reserved.Journal of Pharmaceutical and Biomedical Analysis 10/2014; 102C:468-475. DOI:10.1016/j.jpba.2014.10.004 · 2.83 Impact Factor