• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.99-99
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• 2018

The major aim of this work is the research of secondary metabolites isolated from the aerial parts of Petasites japonicus. The plant material is extracted with a polar solvent, which is 95% by volume methanol at room temperature. The concentrated extract was partitioned as EtOAc, n-BuOH, and $H_2O$ fractions. From the EtOAC and n-BuOH fraction, two bakkenolides and two caffoylquinic acid were isolated using the Diaion HP-20, silica gel, ODS-A, and Sephadex LH-20 column chromatographies. According to the results of the results of physico-chemical and spectroscopic data including NMR, MS and UV. The chemical structures of the compounds were respectively determined as bakkenolide B (1), bakkenolide D (2), 1,5-dicaffeoylquinic acid (3), and 5-O-caffeoylquinic acid (4). These results suggest that the compounds isolated from the aerial parts of this plant were almost identical with known components of Petasites japonicus. However, it is necessary to investigate more about the difference of amounts of constituents according to harvest area and time.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.11
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• pp.1610-1616
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• 2016

This study aimed to establish an optimal extraction process and high performance liquid chromatography-ultraviolet (HPLC-UV) analytical method for determination of 3,5-dicaffeoylquinic acid (3,5-DCQA) as a part of materials standardization for the development of a xanthine oxidase inhibitor as a health functional food. The quantitative determination method of 3,5-DCQA as a marker compound was optimized by HPLC analysis using a Luna RP-18 column, and the correlation coefficient for the calibration curve showed good linearity of more than 0.9999 using a gradient eluent of water (1% acetic acid) and methanol as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 320 nm. The HPLC-UV method was applied successfully to quantification of the marker compound (3,5-DCQA) in Aster glehni extracts after validation of the method with linearity, accuracy, and precision. Ethanolic extracts of A. glehni (AGEs) were evaluated by reflux extraction at 70 and $80^{\circ}C$ with 30, 50, 70, and 80% ethanol for 3, 4, 5, and 6 h, respectively. Among AGEs, 70% AGE at $70^{\circ}C$ showed the highest content of 3,5-DCQA of $52.59{\pm}3.45mg/100g$ A. glehni. Furthermore, AGEs were analyzed for their inhibitory activities on uric acid production by the xanthine/xanthine oxidase system. The 70% AGE at $70^{\circ}C$ showed the most potent inhibitory activity with $IC_{50}$ values of $77.01{\pm}3.13{\sim}89.96{\pm}3.08{\mu}g/mL$. The results suggest that standardization of 3,5-DCQA in AGEs using HPLC-UV analysis would be an acceptable method for the development of health functional foods.

• Korean Journal of Food Science and Technology
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• v.51 no.1
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• pp.81-89
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• 2019

This study was performed to confirm the influence of chlorogenic acid (CGA) and 3,5-dicaffeyolquinic acid (3,5-diCQA) intake on problems caused by high-fat diet. CGA was more effective in suppressing weight gain than 3,5-diCQA. In contrast, 3,5-diCQA was more effective in improving glucose tolerance than CGA. In the biopsy, it was confirmed that CGA inhibited visceral fat and liver fat accumulation. 3,5-diCQA also inhibited visceral fat accumulation, but 3,5-diCQA increased liver fat accumulation. The liver fat accumulation induced oxidative stress, but 3,5-diCQA reduced oxidative damage through its antioxidant activity. The increased liver fat accumulation was because a 3,5-diCQA greatly increased Akt phosphorylation and decreased AMPK phosphorylation in the liver. Consequently, CGA was effective in alleviating the problems caused by high-fat diets, while maintaining normal balance. 3,5-diCQA also showed a positive effect on problems caused by high-fat diets, but it increased liver fat accumulation and thereby had negative consequences.

• Journal of the Korean Magnetic Resonance Society
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• v.22 no.2
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• pp.26-33
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• 2018

Xanthium sibiricum is used as a traditional folk medicine for the treatment of cancer, fever, headache, nasal sinusitis, and skin pruritus. This study aimed to identify components from Xanthium sibiricum extracts using an SPE-800MHz NMR-MS hyphenated system. The simultaneous acquisition of MS and NMR spectra from the same chromatographic peaks significantly increases the depth of information acquired for the compound and allows the elucidation of structures that would not be possible using MS or NMR data alone. LC -NMR analysis was conducted using a HPLC separation system coupled to 800 MHz spectrometer equipped with a cryoprobe, and a SPE unit was used to automatically trap chromatographic peaks using a HPLC pump. LC-MS analysis was conducted with a Q-TOF MS instrument using ESI ionization in the negative ion mode. Using the hyphenated analysis, several secondary metabolites were identified, such as 3',5'-O-dicaffeoylquinic acid, 1',5'-O-dicaffeoyl- quinic acid, and ethyl caffeate. These results demonstrate that the SPE-800MHz NMR-MS hyphenated system can be used to identify metabolites within natural products that have complex mixtures.

• Korean Journal of Pharmacognosy
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• v.48 no.3
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• pp.243-247
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• 2017

To find the anti-Alzheimer's activity of Cacalia hastata var. orientalis (Compositae), caffeoylquinic acids were analyzed in this plant by HPLC and the inhibitory activities of the extract on cholinesterases including acetylcholinesterase (AChE) and butyrylcholinesterse (BChE) and peroxynitrite ($ONOO^-$) were assayed. Sum of caffeoylquinic acids in the 80% MeOH extract was quantitatively higher in leaves (35.8% of the extract) than in stems (3.7%). The compound, 3,5-dicaffeoylquinic acid, was contained in the highest amount in the leaf (44.32 mg/g dry weight). The $IC_{50}s$ of the 80% MeOH extract were shown to be $67.45{\mu}g/ml$ for AChE, and $8.59{\mu}g/ml$ (for $ONOO^-$), respectively, suggesting that the leaves would have the anti-Alzheimer's activity due to the high content of caffeoylquinic acids.

• The Korea Journal of Herbology
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• v.30 no.3
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• pp.19-24
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• 2015

Objectives : HPL-1, a novel herbal medicine which is composed of five herbs such as Kalopanacis Cortex, Chaenomelis Fructus, Raphani Semen, Atractylodis Rhizoma and Pulvis Aconiti Tuberis Purificatum, was developed for treatment of osteoarthritis. This study is aimed to develop analytical method for consistent quality control of HPL-1 and validate chromatographic method. Methods : Chromatographic analysis was performed using ultra-high performance liquid chromatography - diode array detector (UHPLC-DAD) equipped with RP-amide column, column oven, and auto sampler. Marker compounds [protocatechuic acid, chlorogenic acid, liriodendrin, 3,5-dicaffeoylquinic acid, ${\beta}$-D-(3-O-sinapoyl)-fructofuranosyl-$\alpha$-D-(6-O-sinapoyl)glucopyranoside and benzoylmesaconine] were separated by step gradient elution of acetonitrile and 0.1% phosphoric acid/water. The method validation was evaluated by quantitative validation parameters of linearity, accuracy, precision, limit of detection (LOD) and limit of quantification (LOQ) according to KFDA guideline.Results : An optimized method for six marker compounds in HPL-1 was established by UHPLC-DAD. The correlation coefficient (R²) with each calibration curve was greater than 0.99. The LOD and LOQ were within the range of 0.008-0.090 and $0.023-0.274{\mu}g/mL$, respectively. The relative standard deviation (RSD) of intra- and inter-day variability were less than 4.0%. The result of recovery test was range from 93.3-106.3% with RSD < 4.0%.Conclusions : These results suggest that the quantitative UHPLC method is precise, accurate, effective for quality evaluation of HPL-1. The method may also contribute to improve quality of crude drug preparations used for treatment of various diseases.

• YAKHAK HOEJI
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• v.52 no.6
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• pp.446-451
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• 2008

A high-performance liquid chromatography (HPLC) with diode array detector (DAD) and electrospray ionization mass spectrometry (ESI-MS) was established for the simultaneous determination of chlorogenic acid (1), sweroside (2), luteolin-7-O-glucoside (3), (E)-aldosecologanin (4) and 3,5-dicaffeoylquinic acid (5) from Lonicera joponica flower buds. The optimal chromatographic conditions were obtained on an ODS column (5 ${\mu}m$, 4.6${\times}$150 mm) with the column temperature $25^{\circ}C$. The mobile phase was composed of (A) water with 0.1% formic acid and (B) acetonitrile with 0.1% formic acid using a gradient elution, the flow rate was 0.3 ml/min. Detection wavelength was set at 250 nm. All calibration curves showed good linear regression ($r^2$>0.994) within test ranges. The developed method provided satisfactory precision and accuracy with overall intra-day and inter-day variations of 0.05${\sim}$1.95% and 0.15${\sim}$2.26%, respectively, and the overall recoveries of 97.71${\sim}$103.65% for the five compounds analyzed. The verified method was successfully applied to quantitative determination of the three types (phenolic compounds, iridoids and flavonoids) of bioactive compounds in 21 commercial L. japonica flower buds samples from different markets in Korea and China. The analytical results demonstrated that the contents of the five analytes vary significantly with sources.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.11
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• pp.1701-1708
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• 2014

Phenolic compounds, physiological properties, and digestive enzyme inhibitory effect of 70% ethanol extracts from Aster scaber with different extraction methods (stirrer extraction, SE; reflux extraction, RE; autoclave extraction, AE; low temperature high pressure extraction, LTPE; ultrasonification extraction, USE) were investigated. Total polyphenols and flavonoids contents in LTPE were significantly higher than those of other extracts. The amount of substances related to cynarin (1,3-O-dicaffeoylquinic acid) was highest in USE (34.34 mg/g), followed by LTPE (33.83 mg/g), RE (32.27 mg/g), AE (25.40 mg/g), and SE (18.17 mg/g). Chlorogenic acid (5-O-caffeoylquinic acid) and astragalin (kaempferol-3-O-glucopyranoside) were highest in AE and LTPE, respectively. Xanthine oxidase, angiotensin- I converting enzyme, 3-hydroxy-3-methylglutaryl coenzyme A reductase and acetylcholin esterase inhibitory activities of LTPE and USE at a concentration of 50 mg% (w/v) were somewhat higher than those of other extracts. The ${\alpha}$-amylase, ${\alpha}$-glucosidase, trypsin and lipase activities showed the same tendency as physiological properties (concentration of 500 mg%, w/v). Additionally, there was significantly higher or slightly lower inhibitory activity compared to the control group. These results suggest that extracts from Aster scaber have potential to act as functional materials, and LTPE and USE are superior for the enhancement of biological activity.

• Journal of Naturopathy
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• v.12 no.1
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• pp.7-15
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• 2023

Background: As a result of analyzing the components of wild Conyza canadensis, it contains physiologically active ingredients, so it is necessary to identify the compound. Purposes: It was to study the compound's molecular structure; a previous study showed that C. canadensis contains antioxidant substances. Methods: The ultrasonic pulverized lysate of C. canadensis stem and leaves was first extracted with 90% methanol and then five organic solvents. Next, the extracts was fractionated by HPLC, LC/MS chromatography, and NMR analyzers identified the molecular structure. Results: 100 g of dry C. canadensis was sonicated in 90% methanol and concentrated under reduced pressure to 11.96 g of a crude extract. Then, this crude was extracted with five types of solvents to obtain 123.8 mg of n-hexane, 448.2 mg of dichloromethane, 1047.7 mg of ethyl acetate (EA), 2563.8 mg of butanol, and 7.04 g of water. The EA extracts were fractionated by LC-MS and then re-fractionated to obtain F1 to F20. Next, the F15 was further fractionated to obtain nine fine fractions. Finally, the F17 fraction was re-fractionated to obtain ten fine fractions. As a result of LC-MS and NMR spectrometer analysis of the F15-7, the structure of this compound was confirmed as 3,5-dicaffeoylquinic acid. As a result of examining the structures of the F17-4 and F17-5 fractions, Quercetin-3-o-β-galactose was identified. In addition, the form of the F17-10 was confirmed to be 1,3,4-tri-caffeoylquinic acid. Conclusions: This study demonstrated that C. canadensis contained phenolic antioxidants, and its utilization may be expected.

• Korean Journal of Environmental Agriculture
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• v.37 no.4
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• pp.260-267
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• 2018

BACKGROUND: In this study, shoots of Aralia elata and Kalopanax pictus which belong to the Araliaceae family were analyzed using UPLC-DAD-ESI(+)-QToF/MS to characterize of individual phenolic acids. METHODS AND RESULTS: Total thirteen phenolic acids were identified, and nine hydroxycinnamic acid derivatives have been identified for the first time in shoots of Aralia elata and Kalopanax pictus. For total phenolic acid content (mg/100g dry weight), shoots of Aralia elata and Kalopanax pictus showed 754.8 and 845.3 mg/100g, respectively. 5-O-Caffeoylquinic acid (49%) and 3,5-di-O-caffeoylquinic acid (44%) were found as major phenolic acids in Aralia elata, while 5-O-caffeoylquinic acid (91%) was a major component in Kalopanax pictus. CONCLUSION: On comparing the two plants, it was considered that the biosynthesis of 3,5-di-O-caffeoylquinic acid can be affected by 5-O-caffeoylquinic acid in Aralia elata.