• The Korean Journal of Food And Nutrition
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• v.30 no.1
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• pp.96-104
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• 2017

To obtain the immunomodulating polysaccharide from chaga mushroom (Inonotus obliquus sclerotia, IO), crude polysac- charide fractions (IO-M-CP and IO-CP, respectively) prepared from hot-water extract (IO-W) of I. obliquus by EtOH precipitation after MeOH reflux or not. After IO-W was re-dissolved in water followed by EtOH addition in the case without MeOH reflux, EtOH mixture was fractionated into EtOH-soluble (IO-E) and crude polysaccharide (IO-CP). In the meanwhile, MeOH-soluble fraction (IO-M) was separated from IO-W after MeOH reflux. The residue was dissolved in water and was added by EtOH, and then EtOH mixture was also fractionation into EtOH-soluble (IO-M-E) and crude polysaccharide (IO-M-CP). As a result of the macrophage stimulating activity of these fractions, IO-CP and IO-M-CP showed significantly increased cell proliferation and cytokines production than IO-W. Particularly, IO-M-CP promotes the production of IL-12 more than IO-CP. In the splenocytes proliferating activity and intestinal immune system modulating activity through Peyer's patch, both of 2 crude polysaccharide fractions were significantly promoted in cell proliferation and cytokines production than IO-W, and IO-M-CP was more potent than IO-CP in IL-2 production from splenocytes and GM-CSF production ($10{\mu}g/mL$) in Peyer's patch cells. In addition, immunomodulating polysaccharide fractions (IO-M-CP and IO-CP) prepared from IO-W by EtOH precipitation with or without EtOH reflux showed no significant difference in the chemical composition and component sugar. These results suggested that MeOH reflux might exclude low-molecular weight materials from IO-W and consequently increase the immunomodulating activity of IO-M-CP. Therefore, it was confirmed that immunomodulation of polysaccharide prepared from hot-water extract of chaga mushroom was enhanced by fractionation including MeOH reflux and EtOH precipitation.

• Korean Journal of Food Science and Technology
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• v.51 no.2
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• pp.103-108
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• 2019

This study was conducted to select the optimal extraction method of codonopsis lanceolata saponin. To investigate the lancemasides content depending on each extraction method, various extractions were performed: reflux (methanol and butanol), hot water, as well as ultrasonic bath (40 kHz; continuous irradiation/interval irradiation) and ultrasonicator (20 kHz) extractions. From the result, the overall lancemasides content were the highest in ultrasonic bath (MeOH; continuous irradiation) extraction, followed by ultrasonic bath (water; continuous irradiation)>ultrasonic bath (MeOH; interval irradiation)>ultrasonicator (MeOH)>hot water>MeOH reflux>BuOH reflux extractions in that order. Sample drying method prior to ultrasonic bath extraction was more effective shade drying than freeze drying. Effective duration and temperature of extraction was 2 hr at $64^{\circ}C$. And ingredient change diverted from aster saponin Hb to lancemasides was identified by extraction condition such as extraction time and temperature.

• Korean Journal of Medicinal Crop Science
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• v.16 no.6
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• pp.446-454
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• 2008

An advanced extraction method by ultrasonic extraction with applied solid phase extraction (SPE) has been developed for the determination of simultaneous eight major ginsenosides, namely ginsenosides Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, and Rd in the root of Panax ginseng. Four extraction methods including n-BuOH reflux extraction (Method A), 70% EtOH reflux extraction (Method B), 50% MeOH reflux extraction with SPE (Method C), and 50% MeOH ultrasonication with SPE clean-up process (Method D) were investigated for the determination of eight major ginsenosides. Total contents of ginsenosides were highest by extraction of Method C as $2.408{\pm}0.011%$. However, Method D was evaluated as relatively simpler and more efficient method due to short extraction time, small solvent consumption and less expensive, compared to conservative reflux method. Ginsenosides were also satisfactorily separated with good resolution and the accuracy range was between 1.05 and 4.06% as relative standard deviation (RSD) by Method D. SPE condition and HPLC condition were further optimized for determination of eight major ginsenosides by the ultrasonic extraction method. Conclusively, ultrasonic extraction of 2 g sample of ginseng using ultrasonic bath and 1 loading for SPE was evaluated as proper condition for extraction of ginseng.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.3
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• pp.356-362
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• 2010

When pine (Pinus densiflora) needles were fractionated into cold water (PD-CW), hot water (PD-HW) and methanol extract (PD-M), PD-M showed potent simulating activity (1.19-fold of the saline control) for proliferation of bone marrow cells mediated by Peyer's patch cells in vitro. MeOH extracts were prepared by homogenization, stirring or reflux to identify the method of methanol extraction, and MeOH extract by reflux method showed significantly highest intestinal immune system modulating activity (1.30-fold) in vitro. The intestinal immune system modulating effect of orally administered PD-M fractionated from pine needles also were studied in mice. Analyzing intestinal immune system modulating activity mediated Peyer's patch cells from C3H/He mice which had been fed with PD-M at different doses for 7 days, 1.0 g/kg of BW/day indicated that the bone marrow cells had proliferated (3.65-fold of 3% EtOH administered group). In addition, the amounts of IL-6 in the culture supernatant of Peyer's patch cells at 1.0 g/kg of BW/day were increased (1.13-fold) whereas the production of GM-CSF was not dose dependent. These results indicate that oral administration of PD-M enhances the secretion of hematopoietic growth factors such as GM-CSF and IL-6 from Peyer's patch cells, and these cytokines also act on modulator of bone marrow cell proliferation.

• Korean Journal of Pharmacognosy
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• v.36 no.3 s.142
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• pp.177-185
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• 2005

For the purpose of the quality control of Poria cocos, three major compounds were isolated and identified as pachymic acid (1), $3{\beta}-hydroxylanosta-7,9(11),24-trien-21-oic$ acid (2), and dehydroeburicoic acid (3). The optimal extraction conditions for the quantification of pachymic acid its analogues were the 3 hours of reflux with 15g of P. cocos in 100ml 95% ethanol. HPLC conditions were as follows: Column; ZORBAX Eclipse XDB C18 $(4.6{\times}250\;mm,\;Agillent)$, mobile phase; 1% HOAc in 70% $MeOH{\rightarrow}1%$ HOAc in 100% MeOH for 25 min, then 1% HOAc in 100% MeOH for 15 min, detector, ELSD, flow rate; 1ml/min. The mean contents of 1,2, and 3 in Poria cocos cultivated at 18 different site were $0.65{\pm}0.19,\;0.88{\pm}0.72,\;and\;0.84{\pm}0.54\;mg/g$, respectively, and values might be the guide line for the quality control of P. cocos.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.53 no.1
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• pp.110-117
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• 2008

Soybeans (Glycine max) with a black seed coat have been widely utilized as food source and as a medicinal herbs in Korea. The pigmentation in the seed coat of black soybean is due to accumulate anthocyanins in the epidermis palisade layer. The anthocyanin content and composition of the black soybean seed coat are considered as a standard. of quality evaluation of black soybean. The main objective of this study was to investigate the optimal condition for an extraction method of anthocyanins and compare anthocyanin quantity and composition within black soybean varieties and germplasms. In the test of extraction solvent, absorbance at 530 nm and Hunter's a value were increased as increasing the concentration of MeOH, but Hunter's Land b values were the exact opposite of absorbance and Hunter's a values. There was no significant difference for anthocyanin contents from 1% HCl - $H_2O$ to 1% HCl - 80% MeOH. In the aspects of anthocyanin contents and HPLC peak resolution, 1% HCl - 20% MeOH extraction solution was the most suitable solvent. Among the 5 kinds of extraction method using 1% HCl - 20% MeOH solution, the anthocyanin contents of room temperature extraction at 72 h was the highest among the methods. High extraction temperature, sonication and reflux method influenced on the decrease of anthocyanin contents because of breakdown of anthocyanins. There was no significant difference for extraction time between 12 h and 24 h. However, the optimal extraction condition were at room temperature for 12 h. The anthocyanin contents in seed coats of black soybean were determined on the basis of HPLC peak area at 530 nm. Ten black soybean varieties and germplasms were tested with optimal conditions founded in this study. On the basis of antocyanin component, these can be classified into three groups; C3G, C3G + D3G and C3G + D3G + Pt3G. The total anthocyanin content in seed coats ranged from 1.58 to 10.62 mg/g of seed. The total anthocyanin content of the variety "Geomjeongol" was about 7 times higher than that of variety "Heugchong". Information for extraction method and diversity in antocyanin of soybean seed coats can be used for future research for germplasm evaluation and development of high quality black soybean varieties.

• Childhood Kidney Diseases
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• v.2 no.2
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• pp.152-160
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• 1998

Purpose : Vesicoureteral reflux (VUR), which can result in renal failure in children, is expected to resolve spontaneously in mild cases, but severe cases require surgical correction, posing dilemma in deciding the right measures in some cases. Therefore, to delineate the factors affecting spontaneous resolution of VUR and to apply them in assessing the prognosis, we reviewed the patients with VUR who had been treated medically. Patients and Methods : 32 children (49 renal units) with reflux who had been admitted during the 5-year period from Jan '92 to Dec '96 were classified into 4 groups (Resolved, Improved, Unchanged, and Worsened) and analyzed. Results : 1) 25 were boys and 7 girls. The age at diagnosis ranged from 13 days to 9 years (mean $24.6{\pm}11.4$ mo) 2) Among 49 refluxing ureters, 4 were below 1 me of age,21 between 1 me to 1 yr, 16 between 1 yr to 6 yr, 8 beyond 6 yr. Two belonged to Grade I, 16 to Crade II, 17 to Grade III, 12 to Grade IV, and 2 to Grade V. 3) Spontaneous resolution rates of reflux were 100, 81, 47, 8, 0$\%$ for each Grade, respectively. Resolution plus improvement rates in Grade III and IV were 71 and $50\%$, each. 4) Resolution rates in relation to the age at diagnosis were $100\%$ for below 1 mo, $48\%$ between 1 mo to 1 yr, $56\%$ between 1 to 6 yr, and $13\%$ beyond 6 yr. 5) Of 15 unilateral refluxing ureters, 14 ($93\%$) resulted in resolution. Of 34 bilateral refluxing ureters, 11 ($32\%$) resolved spontaneously 6) Resolution occurred within 1 year in 20 units of 24 renal units in regular follow-up. Conclusion: The lower the initial grade and the younger the patient, the sooner the reflux resolved. Resolution was better when VUR was unilateral than bilateral. Long-term regular follow-up is essential because even the cases falling in Grades III and IV have high rates of spontaneous resolution and improvement with medical treatment.

• Korean Journal of Pharmacognosy
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• v.47 no.1
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• pp.12-17
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• 2016

In this study, the purification of the MeOH extract from the stems and leaves of Pourthiaea villosa var. brunnea using column chromatography furnished a main compound, lyoniside. The structure was elucidated on the basis of $^1H$ and $^{13}C$ NMR spectroscopic data. Quantitative analysis of lyoniside was conducted by HPLC method and the highest content of lyoniside was found in 50% MeOH reflux extraction. To investigate the anti-inflammatory effect of the lyoniside, we measured nitric oxide and pro-inflammatory cytokines such as TNF-${\alpha}$, IL-$1{\beta}$, IL-6 levels in lipopolysaccharide-induced murine macrophage cell line RAW 264.7. As a results, lyoniside decreased the level of nitric oxide and IL-6 in concentration dose dependent manner in RAW 264.7 cells.

• Journal of Pharmacopuncture
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• v.15 no.4
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• pp.52-65
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• 2012

Objectives: Toad venom, called Chan-Su, is a traditional Oriental medicine secreted from the auricular and the skin glands of the Bufo bufo gargarizanz Cantor or B. melanosticus Schneider and has been widely used in China, Korea and other parts of Asia for the treatment of pain, heart conditions, and cancer. We examined the concentrations of the main chemical constituents within a commercially available toad venom product and compared the levels for different extraction methods. Methods: Toad venom was extracted using either cold or hot water, ethanol (EtOH), methanol (MeOH), or ethyl acetate (EtOAc), was fractionated using precipitation or reflux, and was then analyzed using thin layer chromatography (TLC), high-performance liquid chromatography (HTLC), and liquid chroma-tography - mass spectrometry (LC-MS). Individual components were identified by comparisons of the retention times, the ultraviolet spectra, and mass spectras and differences in chemical constituents for different solvents and extraction methods are presented. Results: Components with authentic standards, including serotonin and bufodienolides (cinobufagen, bufalin, cinobufalin, and resibufogenin), were detected. The water extract of toad venom contained the greatest amount of serotonin ($75.7{\pm}0.1$ mg/g), but very small amounts of bufodienolides ($3.8{\pm}0.0$ mg/g). In contrast, the use of MeOH or EtOH extraction solutions resulted in 5-26 times higher concentrations of bufodienolides, with only trace amounts of serotonin. The relative and the absolute concentrations of the component also varied based on the extraction method; i.e., EtOH extracts yielded the greatest total amounts of bufodienolides, and EtOAc precipitation had the lowest amounts of bufodienolides. Conclusions: Toad venom consists of serotonin and several bufodienolides, and the choice of solvent to extract chemical the constituents is important as a way to enrich the purported active components for treating different conditions.

• Textile Coloration and Finishing
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• v.14 no.4
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• pp.229-239
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• 2002

Colorants were extracted from the heartwood of sappan lin. with MeOH under reflux, and the concentrate or the powder of dye was prepared by low pressure concentration method using suitable organic solvent. Various components were isolated from sappan wood, and the chemical structure and mechanism of compound having the excellent antibacterial and deodorization properties were analyzed. The results obtained are as follows ; The seventeen components of sappan wood were seperated by HPLC chromatography, and the five components among them were existed more than 6% and the other components were existed lower than 0.6%. The resolving powers of the non-polar solvent and polar solvent systems were evaluated by their ability to resolve the samples. It showed that chloroform-methanol-water(800:150:10) system has the best resolving power. Although the seperation rate is very slow, polyamide C-100 column chromatography gives a clear seperation of sappan wood. On the basis of the spectrometric data such as IR, UV, $GC-Mass,\;^1H-NMR,\;^{13}C-NMR\;and\;^1H-^{ 13}C-NMR$, the chemical structure of compound haying the excellent antibacterial and deodorization properties was established as brazilin containing the functional groups such as two quaternary carbon, one benzyl carbon, methylene contiguous to oxygen and methylene caused by oxygen atom.