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한국생약학회> 생약학회지> 곰보배추에 함유된 6-Hydroxyluteolin 7-O-Glucoside 분석을 위한 HPLC/DAD 분석법 밸리데이션

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곰보배추에 함유된 6-Hydroxyluteolin 7-O-Glucoside 분석을 위한 HPLC/DAD 분석법 밸리데이션

HPLC/DAD Method Validation of 6-Hydroxyluteolin 7-O-Glucoside Analysis from Salvia plebeia

이학동 ( Hak-dong Lee ) , 바헤 ( Leo Adrianne Paje ) , 최중원 ( Jungwon Choi ) , 김주리 ( Juree Kim ) , 유아람 ( A Ram Yu ) , 배민정 ( Min-jung Bae ) , 이상현 ( Sanghyun Lee )
  • : 한국생약학회
  • : 생약학회지 52권3호
  • : 연속간행물
  • : 2021년 09월
  • : 186-191(6pages)
생약학회지

DOI


목차

Materials and Methods
Results and Discussion
Acknowledgments
References

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We developed a method to identify and quantify 6-hydroxyluteolin 7-O-glucoside in the powder of Salvia plebeia (PS) using high-performance liquid chromatography coupled with diode array detector (HPLC/DAD) and equipped with reverse-phase INNO C18 column. The analytical method was optimized and validated using novel parameters. The obtained values for the limits of detection and quantification were 3.60 and 10.90 μg/mL, respectively. Calibration curve showed good linearity in the concentration range tested (0.00625-0.1 mg/mL, r 2 = 1.0000), high accuracy (96.2-101.4%), and precision values (RSD ≤ 0.27%). Our analysis support the use of our method for accurately identifying and quantifying 6-hydroxyluteolin 7-Oglucoside from PS in routine analyses and large-scale extraction processes for content determination.

UCI(KEPA)

간행물정보

  • : 의약학분야  > 약화학
  • : KCI등재
  • : SCOPUS
  • : 계간
  • : 0253-3073
  • : 2288-9299
  • : 학술지
  • : 연속간행물
  • : 1970-2021
  • : 3353


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발행기관 : 한국생약학회 간행물 : 생약학회지 52권 4호 발행 연도 : 2021 페이지 : pp. 203-207 (5 pages)

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Four compounds were isolated from 90% ethanol extract of the stem of Clematis heracleifolia. On the basis of spectral data, the structure of isolated compounds were identified as sitosterone (1), β-sitosterol (2), scoparone (3) and octadecanoyl caffeate (4), respectively. Sitosterone (1) and octadecanoyl caffeate (4) are isolated from this plant for the first time.

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3팔손이나무(Fatsia japonica) 줄기의 성분연구

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발행기관 : 한국생약학회 간행물 : 생약학회지 52권 4호 발행 연도 : 2021 페이지 : pp. 212-218 (7 pages)

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Fatsia japonica is grown wild to Eastern Asia, including Korea, Japan, and Taiwan and it is known as ornamental plant, and it is also known that pharmacological action. In this study, we have selected the stem of F. japonica with consideration about biological activities and amount of yield. In addition, four compounds (1-4) were isolated from the stem of F. japonica. Extensive spectroscopic and chemical studies established the structures of these compounds as maltose (1), begoniifolide A (2), leiyemudanoside B (3), leonticin F (4). All of the compounds were investigated for their anti-inflammatory, antineuroinflammatory, and neuro-protective effects on RAW264.7, BV2, and HT22 cells. However, among four compounds, there were no effects by maltose (1), begoniifolide A (2), leiyemudanoside B (3), leonticin F (4) on the anti-inflammatory, anti-neuroinflammatory, and neuro-protective action. This is the first report on the isolation of maltose (1), begoniifolide A (2), leiyemudanoside B (3), leonticin F (4) from the stem of F. japonica. Begoniifolide A (2), leiyemudanoside B (3), leonticin F (4) were isolated for the first time from this plant. It might be necessary to continue the further studies to find the biological active compounds isolated from the stem of F. japonica.

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4벤조피렌 기준 미설정 한약재의 오염도 조사 및 위해도 결정

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By analysing the benzo(a)pyrene concentration using HPLC-FLD and LC-MS/MS, pollution levels of herbal medicines without permitted benzo(a)pyrene specification were investigated. Average benzo(a)pyrene concentrations were 38.30, 37.46 and 21.22 μg/kg for Amomi Tsao-ko Fructus, Mume Fructus and Coptidis Rhizoma, respectively, which are 4 to 7 times higher than maximum permitted benzo(a)pyrene concentration of Rehmanniae Radix Preparata and Rehmanniae Radix, i.e. 5.0 μg/kg. Proportion of detected samples exceeding 5.0 μg/kg benzo(a)pyrene concentration was 22% for Cimicifugae Rhizoma and Scrophulariae Radix, 44% for Forsythiae Fructus, 67% for Mume Fructus, 100% for Amomi Tsao-ko Fructus and Coptidis Rhizoma, and collectively 29% (36 out of 125 samples) in average. In terms of risk characterization results, human exposure of benzo(a)pyrene were 7.96, 3.49 and 1.61 ng/kg b.w./day and the margin of exposure(MOE) were 1.25 × 104, 2.86 × 104 and 6.20 × 104 for Mume Fructus, Amomi Tsao-ko Fructus and Coptidis Rhizoma, respectively. MOE banding of those herbal medicines was categorized to 'low concern'. However, considering that human exposure of benzo(a)pyrene for food is legislated to 1.4~2.5 ng/kg b.w./day, it was urgent to set up the guideline of benzo(a)pyrene in herbal medicines.

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5자외선 조사에 의해 노화된 인간각질형성세포에서 구멍쇠미역 추출물의 항노화 효능

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This research was carried out to investigate the moisturizing effects of Agarum cribrosum extract on normal human epidermal keratinocytes (NHEKs). Moisturizing effects of A. cribrosum extract on NHEKs were measured by quantitative realtime RT-PCR to verify the gene expressions related to skin hydration, hyaluronic acid (HA)-ELISA to detect HA production, and cell viability assays. A. cribrosum extract increased the mRNA levels of the AQP3 and HAS2 genes and HA production in NHEKs. On the other hand, A. cribrosum extract decreased the mRNA level of the KRT1 and KRT10 genes known as differentiated keratinocyte marker in NHEKs. This research showed the moisturizing effects of A. cribrosum extract. The results indicate that A. cribrosum extract can be a potent functional ingredient for skin hydration and anti-aging products. Further study is warranted regarding the use of A. cribrosum extract to develop not only cosmetics but also food and medicine.

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Dermal papilla cells (DPCs) are present throughout the hair cycle and play an essential role in hair cycle and hair growth. In this study, we investigated the effect of Carex dispalata on the activation of anagen pathway in DPCs. C. dispalata extract increased the proliferation of DPCs and induced changes in the levels of cell cycle-related proteins. To elucidate the mechanism by which C. dispalata extract stimulates the anagen pathway related to the proliferation of DPCs, we evaluated the effect of C. dispalata extract on the activation of Akt signaling. The increase in the level of phospho-Akt by C. dispalata extract was inhibited by PI3K inhibitor (wortmannin). Wortmannin reduced the effects of C. dispalata extract on the levels of cell cycle-related proteins and proliferation of DPCs. C. dispalata extract increased the levels of Wnt/β-catenin proteins. Wnt/β-catenin inhibitor ( XAV939) inhibited changes in cell cycle, cell cycle-related proteins, Wnt/β-catenin proteins, and proliferation induced by C. dispalata extract. C. dispalata extract increased the level of autophagy protein (LC3I/II), and this change was inhibited by XAV939. These results suggest that C. dispalata extract can activate PI3K/Akt, Wnt/β-catenin, and autophagy pathways in DPCs to induce cell proliferation, and thereby promote hair growth phase.

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Artemisia frigida Willd (AW, Fringed sagewort), which is widespread in Mongolia, is a well-known medicinal plant as a member of the Compositae family. This study aims to explore the gastroprotective effect of water extract of AW on 150 mM HCl/60% ethanol-induced acute gastritis in 5 week old male ICR mice. Total polyphenols, total flavonoid contents, and anti-oxidant activity in vitro in AW were evaluated. First, the gross area of gastric mucosal damage was measured. Then western blot analysis was conducted to determine the possible mechanisms of action underlying the effects of AW. AW administration decreased gastric mucosal damage. Moreover, the group with AW treatment effectively inhibited nicotinamide adenine dinucleotide phosphate (NADPH) oxidase expression associated with oxidative stress. AW treatment enhanced an anti-oxidant effect through the increase of anti-oxidant proteins. Besides, the increased expressions of inflammatory cytokines induced by nuclear factor-kappa B (NF-κB) activation are alleviated through AW treatment. Taken together, AW exerted a gastroprotective effect against gastric mucosal damage. These results indicate that AW could have the potential used as a natural therapeutic drug for the treatment of acute gastritis.

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Oat, the seeds of Avena sativa L. (Gramineae), is an important dietary staple for people in many countries. Previous studies reported that A. sativa had various pharmacological effects such as anti-inflammatory, antitumor, neurotonic, and antispasmodic activities. In this study, Caenorhabditis elegans model system was used to investigate the antioxidant activity of methanol extract of oat. The ethyl acetate soluble fraction of the oat methanol extract showed the best DPPH radical scavenging activity. The ethyl acetate fraction was measured for the activity of superoxide dismutase (SOD), catalase, and oxidative stress tolerance by using C. elegans along with reactive oxygen species (ROS) level. In addition, to confirm that the regulation of the stress response gene is responsible for the increased stress tolerance of C. elegans treated by the ethyl acetate fraction, SOD- 3 expression was measured using GFP-expressing transgenic worm. As a result, the ethyl acetate fraction increased SOD and catalase activities, and decreased ROS accumulation in a dose-dependent manner. In addition, the ethyl acetate fraction-treated CF1553 worm showed higher SOD-3::GFP intensity compared to the control.

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This study evaluated several in vitro activities including the preliminary assessment of the anti-cancer, anti-inflammatory, and anti-diabetic effects of tree sprout extracts. Chlorogenic, caffeic, and p-coumaric acid contents in tree sprouts were analyzed using high-performance liquid chromatography and an ultraviolet detector. Among the studied tree sprout extracts, the ethanol (EtOH) extract of Rhus verniciflua exhibited the most potent anti-cancer effect by suppressing the cell viability of a human gastric adenocarcinoma cell line, with an IC50 of 7.06 μg/mL. The EtOH extract of Morus alba (MAB) inhibited the secretion of nitric oxide (NO) at a concentration of 100 μg/mL, with an IC50 of 83.44 μg/mL. Moreover, the EtOH extract of Securinega suffruticosa inhibited NO secretion with the lowest IC50 of 54.42 μg/mL. The EtOH extract of Fraxinus mandschurica was the only extract with effective α-glucosidase inhibitory activity. The total content of chlorogenic, caffeic, and pcoumaric acids was the highest in MAB (14.63 mg/g ext.). In conclusion, the beneficial activities of the tree sprout extracts with high phenolic acid content were generally high. Our results provide a theoretical basis for the development of health-promoting supplements and functional foods.

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The heat-mediated reflux apparatus extraction of Paeonia lactiflora Pall. has been widely used as a traditional extraction method. In this paper, the microwave apparatus extraction method of Paeonia Radix was performed and the active ingredients and functional constituents were compared with the reference extraction method. The most effective extraction condition of albiflorin using the microwave was 120℃ in 50% methanol, and paeoniflorin was maximally extracted at 60℃. The reduced level of paeoniflorin molecule at high-pressure and high-temperature extraction condition was caused by the molecular instability. Additionally, the microwave extraction of 50% methanol extracts at 150℃ showed the highest functional constituents determined by in vitro DPPH radical scavenging activity, polyphenol concentration, and tyrosinase inhibition assay. The microwave apparatus was adapted as a rapid, low-cost, and environmentally friendly method to extract active ingredients and the practical extraction conditions of Paeonia Radix can be used in industrial applications.

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6구증구포 맥문동 Ethyl acetate 분획물의 예쁜 꼬마선충 내의 항산화 효과

저자 : 권강무 ( Kang Mu Kwon ) , 김준형 ( Jun Hyeong Kim ) , 양재헌 ( Jae Heon Yang ) , 기별희 ( Byeolhui Ki ) , 황인현 ( In Hyun Hwang ) , 김대근 ( Dae Keun Kim )

발행기관 : 한국생약학회 간행물 : 생약학회지 52권 3호 발행 연도 : 2021 페이지 : pp. 163-169 (7 pages)

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Using the Caenorhabditis elegans model system, the antioxidant activity of methanol extract of the guzeunggupoprocessed Liriope platyphylla F. T. Wang (Liliaceae) tuber was calculated. Between the methanol extracts of guzeunggupo-processed and non-processed L. platyphylla tuber, the processed L. platyphylla tuber showed higher DPPH radical scavenging effect than the non-processed one. The ethyl acetate soluble fraction of the methanol extract of the guzeunggupo-processed L. platyphylla tuber showed the best DPPH radical scavenging activity. The ethyl acetate fraction of the processed sample was measured for the activities of superoxide dismutase (SOD), catalase, and oxidative stress tolerance by using C. elegans along with reactive oxygen species level. In addition, to verify the regulation of the stress response gene is responsible for the increased stress tolerance of C. elegans treated by the ethyl acetate fraction of the processed sample, SOD-3 expression was measured using a transgenic strain (CF1553). Consequently, the ethyl acetate fraction of the processed sample, increased SOD and catalase activities, and decreased ROS accumulation in a dose-dependent manner. Furthermore, the ethyl acetate fraction of the processed sample-treated CF1553 worm showed higher SOD-3::GFP intensity than the control worm.

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7Cyclophosphamide에 의해 유도된 면역 저하 마우스에서 흑미 (Oryza sativa L.) 호분층의 면역 활성 효과

저자 : 이은별 ( Eun Byeol Lee ) , 최지혜 ( Ji-hye Choi ) , 김시현 ( Si Hyun Kim ) , 장환희 ( Hwan Hee Jang ) , 이성현 ( Sung Hyen Lee )

발행기관 : 한국생약학회 간행물 : 생약학회지 52권 3호 발행 연도 : 2021 페이지 : pp. 170-176 (7 pages)

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Anthocyanins are considered as one of the major functional components including cyanidin-3-glucoside and peonidin- 3-glucoside anthocyanins of black rice (Oryza sativa L., BR) aleurone layer (OAL) extract. The immunomodulatory biological effects of OAL has rarely been evaluated. This study has extensively studied the immunomodulatory effects of BR in immunosuppressed mice by CPA. The immunostimulatory effects of BR extract in in vivo at 150 (OAL-1) and 300 mg/kg body weight (OAL-2) for OAL groups were measured and compared to the normal group (CON) ingested with distilled water only or other groups. After the 14 days of oral administration, immune related organ weights, serum immunoglobulin (Ig) A and IgG, and cytokines (IL-12, TNF-α) levels, splenocytes proliferation rate, and NK cell activity were evaluated as immune related biomarkers. The serum IgA and IgG values in the OAL-treated groups increased to the level of the CON. OAL significantly and effectively controlled the IL-12 and TNF-α levels and NK activity compared to the negative control (NC) group. The results suggest that BR aleurone layer with immunomodulatory activities has a great potential as a functional food by itself or as a supplement.

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8만성 역류성 식도염에서 황련과 오수유 혼합물이 식도 점막에 미치는 영향

저자 : 이진아 ( Jin A Lee ) , 오민혁 ( Min Hyuck Oh ) , 신미래 ( Mi-rae Shin ) , 노성수 ( Seong-soo Roh ) , 박해진 ( Hae-jin Park )

발행기관 : 한국생약학회 간행물 : 생약학회지 52권 3호 발행 연도 : 2021 페이지 : pp. 177-185 (9 pages)

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Gastroesophageal reflux disease (GERD) is a gastrointestinal disorder in which stomach contents reflux into the esophagus, causing complications such as mucosal damage. The purpose of this study was to determine the effect of Rhei Rhizoma and Scutellariae Radix mixture (RS) in chronic acid reflux esophagitis (CARE), one of the GERD. After inducing reflux esophagitis through surgery, the group was separated and the drug was administered for 2 weeks; Normal rats (Normal, n=8), chronic acid reflux esophagitis rats (Control, n=8), tocopherol 30 mg/kg-treated chronic acid reflux esophagitis rats (Toco, n=8), Rhei Rhizoma and Scutellariae Radix mixture 100 mg/kg-treated chronic acid reflux esophagitis rats (RSL, n=8), Rhei Rhizoma and Scutellariae Radix mixture 200 mg/kg-treated chronic acid reflux esophagitis rats (RSH, n=8). Gross lesion of esophageal mucosa after RS treatment showed a superior enhancement compared with that of Control group. Additionally, RS significantly decreased the levels of MPO and MDA, effectively inhibited NADPH oxidase, and regulated the expression of the AMPK/ LKB1/NF-κB pathway. Moreover, it significantly increased the expression of tight junction proteins. Taken together, RS not only alleviates inflammation of the esophageal mucosa via the AMPK/LKB1/NF-κB pathway by reducing oxidative stress, but also improves esophageal function by modulating tight junction proteins.

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9곰보배추에 함유된 6-Hydroxyluteolin 7-O-Glucoside 분석을 위한 HPLC/DAD 분석법 밸리데이션

저자 : 이학동 ( Hak-dong Lee ) , 바헤 ( Leo Adrianne Paje ) , 최중원 ( Jungwon Choi ) , 김주리 ( Juree Kim ) , 유아람 ( A Ram Yu ) , 배민정 ( Min-jung Bae ) , 이상현 ( Sanghyun Lee )

발행기관 : 한국생약학회 간행물 : 생약학회지 52권 3호 발행 연도 : 2021 페이지 : pp. 186-191 (6 pages)

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We developed a method to identify and quantify 6-hydroxyluteolin 7-O-glucoside in the powder of Salvia plebeia (PS) using high-performance liquid chromatography coupled with diode array detector (HPLC/DAD) and equipped with reverse-phase INNO C18 column. The analytical method was optimized and validated using novel parameters. The obtained values for the limits of detection and quantification were 3.60 and 10.90 μg/mL, respectively. Calibration curve showed good linearity in the concentration range tested (0.00625-0.1 mg/mL, r 2 = 1.0000), high accuracy (96.2-101.4%), and precision values (RSD ≤ 0.27%). Our analysis support the use of our method for accurately identifying and quantifying 6-hydroxyluteolin 7-Oglucoside from PS in routine analyses and large-scale extraction processes for content determination.

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10제조방법에 따른 당귀수산(當歸鬚散)의 성분분석 및 항염증 효과

저자 : 전영희 ( Young-hee Jeon ) , 남원희 ( Won-hee Nam ) , 임현희 ( Hyun-hee Leem ) , 김세진 ( Se-jin Kim ) , 유병우 ( Byung-woo Yu ) , 손수미 ( Su-mi Son ) , 김명진 ( Myoung-jin Kim ) , 최혜민 ( Hye-min Choi ) , 권현숙 ( Hyun-sook Kwon ) , 김정옥 ( Jung-ok Kim )

발행기관 : 한국생약학회 간행물 : 생약학회지 52권 3호 발행 연도 : 2021 페이지 : pp. 192-201 (10 pages)

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Dangguisu-san (DGSS) is widely known traditional herbal medicinal formula in Korea for treatment of traumatic injury by traffic accident, ecchymosis, abdominal distension and anti-thrombosis of blood. This study was conducted to develop the simultaneous analyze method using high performance liquid chromatography (HPLC) and examine the effect of anti-inflammatory activity of DGSS-dry extract (DGSS-DE) and DGSS-mix extract powder (DGSS-MEP). Physicochemical characteristics of DGSS-DE and DGSS-MEP showed that there is no significant difference in pH, titratable acidity, total soluble solid content and browning degree except for color value (L, a, b). 15 functional constituents of DGSS were identified and the correlation coefficient values of DGSS-DE and DGSS-MEP were conformed 0.950. Also, DGSS-DE and DGSS-MEP significantly decreased the secretion of nitric oxide (NO), prostaglandin E2 (PGE2), interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) through inhibited expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), IL-1β, IL-6, and TNF-α. From these result, DGSS-MEP showed similar chemical composition and anti-inflammatory effect to DGSS-DE. Therefore, DGSS-DE and DGSS-MEP may be useful as potential source of drug to prevent inflammation.

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