Quantification of the Bioactive Components of the Rhizomes of Curcuma wenyujin and Assessment of Its Anti-inflammatory Effect in Benign Prostatic Hyperplasia-1 Cells

서창섭,박은숙,이미영,신현규 한국생약학회 2019 Natural Product Sciences Vol.25 No.3

In this study, the marker compounds of Curcumae Rhizoma (CR) were simultaneously quantified by high-performance liquid chromatography equipped with a photodiode array detector and the anti-inflammatory effects of CR extract and marker compounds in human benign prostatic hyperplasia epithelial-1 (BPH-1) cell lines were investigated. The marker components (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, were separated on Gemini C18 columns (250 mm × 4.6 mm, 5 mm) at 40 oC by using a gradient of two mobile phases eluting at 1.0 mL/min. Prostaglandin E2 (PGE2) levels in Human BPH-1 cells were determined with an ELISA kit. The coefficients of determination in a calibration curve of each analyte were all 0.9997. The limits of detection and quantification of the three compounds were 0.10 – 0.32 mg/mL and 0.30 –0.98 mg/mL, respectively. The content of three compounds, (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, in the CR sample were found to be 5.79 – 5.92 mg/g, 4.72 – 4.86 mg/g, and 1.06 – 1.09 mg/g, respectively. Regarding pharmacological activity against benign prostatic hyperplasia, CR and its components significantly suppressed PGE2 levels of BPH-1 cells. The established analysis method will help to improve quality assessment of CR samples and related products. In addition, CR and its components exhibit anti-inflammatory activity in BPH-1 cells, suggesting the inhibitory efficacy of these compounds against the pathogenesis of BPH.

오매 (烏梅)의 다성분 동시분석 및 항알러지 효과

서창섭,하혜경,이준경,정다영,이진아,신현규 한국생약학회 2012 생약학회지 Vol.43 No.4

The Mume Fructus (MF) has been used for relieves cough, arrests arrest chronic diarrhea, treat fluid depletion, and treat ascariasis in Korea. In this study, a high-performance liquid chromatography (HPLC) method was established for simultaneous determination of six main components of MF. Additionally, we were investigated the anti-inflammatory and anti-allergic effects of MF extract on lipopolysaccharide (LPS)-treated RAW264.7 cells and tumor necrosis factor (TNF)-α/interferon (IFN)-γ-treated HaCaT cells. The analytical column for separation was used a Gemini C18 column maintained at 40o C. The mobile phase consisted of 1.0% (v/v) acetic acid in water (A) and 1.0% (v/v) acetic acid in acetonitrile (B). The flow rate was 1.0 mL/min and the detector was a photodiode array (PDA) set at 280 nm and 320 nm. We evaluated the inhibitory effect of MF extract on the production of inflammatory markers, nitric oxide (NO) and prostaglandin E2 (PGE2) in LPS-stimulated RAW264.7 cells and thymus- and activation-regulated chemokine (TARC/CCL17) in TNF-α/IFN-γ-treated HaCaT cells,respectively. We confirmed the genes expression related with TARC, macrophage-derived chemokine (MDC/ CCL22) and regulated on activation, normal T cell expressed and secreted (RANTES/CCL5) in HaCaT keratinocyte cells by MF extract. The contents of the five compounds in MF were 0.22-1.01 mg/g. Also, the MF extract show inhibition of about 78% and 75% on NO and PGE2 production at the concentration 1000 mg/mL in RAW264.7 cells. MF extract suppressed the hTARC level and genes expression such as TARC, MDC, and RANTES on TNF-α/IFN-γ-treated HaCaT cells.

LC-MS/MS를 이용한 천왕보심단(天王補心丹)의 함량분석 및 항산화 효능 연구

서창섭,김온순,신현규 한국생약학회 2014 생약학회지 Vol.45 No.4

An ultra-performance liquid chromatography-electrospray ionization-mass spectrometer (UPLC-ESI-MS) method was established for the simultaneous quantification of eighteen marker compounds in traditional Korean formula, Cheonwangbosimdan (CWBSD). In addition, we evaluated the antioxidant effects of CWBSD. Eighteen marker components were separated on a UPLC BEH C18 analytical column (2.1?100 mm, 1.7 mm) and kept at 45oC by gradient elution with 0.1% (v/v) formic acid in water and acetonitrile as mobile phase. The flow rate was 0.3 mL/min and the injection volume was 2.0 mL. The antioxidant activities of CWBSD were assessed by measuring free radical scavenging activities on 2,2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The calibration curves of all analytes showed good linearity (correlation coefficient ?0.9937) within the test ranges. The limits of detection and quantification for the 18 marker compounds were 0.01-4.71 ng/mL and 0.03-14.13 ng/mL, respectively. The contents of the 18 compounds in CWBSD extract ranged from none to 1701.00 mg/g. The CWBSD showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction (RC50) against ABTS and DPPH radicals were 149.42 mg/mL and 339.24 mg/mL.

장기보존시험에 따른 쌍화탕의 유통기한 설정

서창섭,김정훈,임순희,신현규 한국생약학회 2012 생약학회지 Vol.43 No.3

The purpose of this study was to estimate the shelf-life of Ssanghwa-tang by long-term storage test. Experiments were conducted to evaluate the stability such as the selected physicochemical, pH, identification, heavy metal, microbiological experiment under a long-term storage test of Ssanghwa-tang. The significant change was not showed in pH, heavy metal,microbiological, identification test and quantitative analysis based on long-term storage test. The contents of albiflorin, paeoniflorin,cinnamic acid, liquiritin, and glycyrrhizin in long-term storage test were 66.8-93.1 µg/mL, 429.0-495.0 µg/mL, 3.8-4.4 µg/mL, 32.0-38.1 µg/mL, and 66.8-71.7 µg/mL, respectively. Shelf-lifes by 5 compounds about 3 lots at room temperature were predicted 21-37, 14-21, and 16-72 months, respectively. The suggested shelf-life would be helpful on the storage and distribution of herbal medicine.

소음인(少陰人) 곽향정기산(藿香正氣散)의 위해물질에 대한 안전성 연구

서창섭,김정훈,황대선,신현규,Seo, Chang-Seob,Kim, Jung-Hoon,Huang, Dae-Sun,Shin, Hyeun-Kyoo 사상체질의학회 2010 사상체질의학회지 Vol.22 No.3

1. Objective: To compare the contents and transfer rate of hazardous substances in crude, washing solution, crude after washing, decoction and remnant after boiling. 2. Methods: The heavy metal contents of each step were measured by inductively coupled plasma mass spectrometer (ICP-MS) and mercury analyzer (MA-2). In order to analyze pesticides in each sample we used simultaneous multi-residue analysis of pesticides by GC/ECD, which was followed by GC/MSD analysis to confirm the identity of the detected pesticide in each sample. In addition, the contents of sulfur dioxide (SO2) were performed by Monier-Williams distillation method. 3. Results: 1) Contents (mg/kg) of heavy metals in decoction of all herbal medicine prescriptions were not detected. 2) Transfer rates (%) of heavy metals from crude to remnant were As (83.3%), Cd (100.0%), Pb (182.6%) and Hg (100.0%). 3) Contents (mg/kg) of residual pesticides were not detected. 4) Transfer rate (%) of sulfur dioxide (SO2) from crude to remnant was 44.2%. 4. Conclusion: Our results showed that boiled herbal medicine prescriptions which we take is safe from the hazardous substances.

장기보존시험에 따른 보중익기탕 전탕팩의 유통기한 평가

서창섭,김정훈,김성실,임순희,신현규 한국생약학회 2013 생약학회지 Vol.44 No.2

The aim of this study was to evaluate the shelf-life of Bojungikgi-tang (Buzhongyiqi-tang in Chinese) by long-term storage test. Experiments were performed to evaluate the stability such as the selected physicochemical, pH, identification,heavy metal, microbiological experiment, and amount of marker compounds under a long-term storage test of Bojungikgi-tang decoction. The significant change was not showed in pH, heavy metal, microbiological, and identification test based on longterm storage test. Furthermore, the HPLC analysis was performed for the determinations of liquiritin, glycyrrhizin, nodakenin,and hesperidin in Bojungikgi-tang by long-term storage test. We were calculated shelf-life of Bojungikgi-tang decoction based on the amount change of four constituents. Consequently, Shelf-life by four compounds at room temperature was predicted 23month. The suggested shelf-life would be helpful on the storage and distribution of herbal medicine.

장기 보존 한약 파우더의 미생물 모니터링

서창섭,신현규,신광수,Seo, Chang-Seob,Shin, Hyeun-Kyoo,Shin, Kwang-Soo 대한한의학방제학회 2011 大韓韓醫學方劑學會誌 Vol.19 No.2

Objectives : This study was carried out to moniter microbial flora on freeze-dried herbal powder and identify isolated bacteria. Methods : We measured the total number of bacteria and fungi in 29 herbal powder which had made according to the guideline of KFDA. For the identification, we observed microscopic properties and carried out polymerase chain reaction(PCR). The purified DNA was analyzed by DNA sequencer. Results : Among the 29 herbal powders, the fungi were detected only one sample as unacceptable range of total aerobic bacteria. Isolated bacteria were identified as Bacillus cereus, B. subtilis, B. megaterium, B. licheniformis, Erwinia tasmaniensis, E. amylovora, and Pantoea agglomerans by 16S rDNA analysis. E. tasmaniensis was observed 20 herbal samples. Conclusions : According to above results, further studies for the effective sterilization of low herbal materials should be needed.

포제에 따른 두충의 지표성분 함량분

서창섭,김정훈,신현규,김병수 한국생약학회 2015 생약학회지 Vol.46 No.2

In this study, we carried out quantification analysis of the six marker components, geniposidic acid, chlorogenic acid, geniposide, pinoresinol diglucoside, liriodendrin, and genipin in the 70% ethanol extracts of non-processed Eucommiae Cortex and processed Eucommiae Cortex using a high-performance liquid chromatography coupled with photodiode array detector. The six components were separated on Gemini C18 column (5 mm, 4.6?250 mm) by the gradient elution with 1.0% (v/v) acetic acid in water and 1.0% (v/v) acetic acid in acetonitrile as mobile phase. The flow rate was 1.0 mL/min and the injection volume was 10 mL. The amount of geniposidic acid, chlorogenic acid, geniposide, pinoresinol diglucoside, liriodendrin, and genipin in non-processed Eucommiae Cortex were 1.31, 0.31, 0.66, 0.46, 0.46, and 0.03%, respectively, while the amount of the six compounds in non-processed Eucommiae Cortex were 0.04-0.78, 0.01-0.14%, 0.05-0.63%, 0.01-0.37%, 0.15-0.42%, and not detected, respectively. After processing treatment, the contents of three iridoids, two lingnan, and one phenylpropanoid decreased in Eucommiae Cortex.

Simultaneous Determination of Seven Compounds in Samsoeum by HPLC-PDA

서창섭,김정훈,황대선,신현규 대한한의학방제학회 2010 大韓韓醫學方劑學會誌 Vol.18 No.1

A HPLC method was developed for simultaneous determination of puerarin, daidzin, liquiritin, naringin, hesperidin, neohesperidin, and glycyrrhizin, to evaluate the quality of SSE. In the present work, simultaneous determination of the seven marker compounds in SSE was validated with respect to linearity, precision, and accuracy. The method will be helpful to improve quality control of SSE.

치자의 포장방법 및 보관기간에 따른 geniposide 함량 분석

서창섭,김정훈,이설림,김성실,김선민,신현규 한약정보연구회 2020 한약정보연구회지 Vol.8 No.2

Gardeniae Fructus (GF) is a traditional herbal medicine listed in “The Korean Pharmacopoeia” and has been used for jaundice, liver disease, and anti-inflammatory. The propose of this study is to establish basic data for setting the shelf life of GF. The content of geniposide, a major compound of GF, was investigated by the packaging method (general, general + silica gel, vacuum, vacuum + silica gel, nitrogen, and nitrogen + silica gel) and storage period (60 months) using high-performance liquid chromatography. As a result, the content of geniposide was initially detected as 4.40%, and then 3.85-5.86% in all packaging methods for 60 months. This result can be used as basic data for quality control and shelf life of GF.