전탕법이 양격산화탕(凉膈散火湯)의 위해물질에 미치는 연구

서창섭,김정훈,하혜경,이미영,이준경,이남헌,이호영,이진아,이설림,신현규,Seo, Chang-Seob,Kim, Jung-Hoon,Ha, Hye-Kyoung,Lee, Mee-Young,Lee, Jun-Kyoung,Lee, Nam-Hun,Lee, Ho-Young,Lee, Jin-Ah,Lee, Sul-Lim,Shin, Hyeun-Kyoo 사상체질의학회 2010 사상체질의학회지 Vol.22 No.2

1. Objectives: To compare the contents of hazardous substances such as crude, washing solution, crude after washing, decoction and remnant. 2. Methods: The heavy metal contents of each step were measured by inductively coupled plasma (ICP) spectrometer and mercury analyzer (MA-2). In order to analyze pesticides in Yanggyeoksanhwa-tang 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) The mean values of heavy metal contents (mg/kg) for each steps in Yanggeoksanhwa-tang were as follows: crude (Pb; 1.87, As; 1.29, Cd; 0.28 and Hg; N.D.), washing solution (Pb; 1.98, As; 1.13, Cd; 0.10 and Hg; N.D.), crude after washing (Pb; 1.90, As; 1.40, Cd; 0.22 and Hg; N.D.), decoction (Pb; 1.90, As; 1.14, Cd; 0.11 and Hg; N.D.) and remnant (Pb; 2.39, As; 1.29, Cd; 0.25 and Hg; 0.01). 2) Contents (mg/kg) of residual pesticides in crude and decoction were not detected. 3) Contents (mg/kg) of sulfur dioxide (SO2) in crude, crude after washing and remnant exhibited 3.00, 2.00 and 2.00 mg/kg, respectively. However, contents of sulfur dioxide in washing solution and decoction were not detected. 4. Conclusions: These Results: will be used to establish a criterion of heavy metals, residual pesticides and sulfur dioxide of each step in Yanggeoksanhwa-tang.

중금속, 잔류농약 및 잔류이산화황의 전탕 전, 후의 농도 변화 - 다빈도 태음인 사상처방을 중심으로 -

서창섭,황대선,이준경,하혜경,천진미,엄영란,장설,김종열,이시우,신현규,Seo, Chang-Seob,Huang, Dae-Sun,Lee, Jun-Kyoung,Ha, Hye-Kyoung,Chun, Jin-Mi,Um, Young-Ran,Jang, Seol,Kim, Jong-Yeol,Lee, Si-Woo,Shin, Hyun-Kyoo 사상체질의학회 2009 사상체질의학회지 Vol.21 No.1

1. Objectives To compare the contents of heavy metals, residual pesticides and sulfur dioxide before/after decoction. 2. Methods The heavy metal contents before/after decoction were measured by Inductively Coupled Plasma Atomic Emission Spectrometry (ICP-AES) and mercury analyzer. In order to analyze pesticides in 3 samples we used simultaneous multi-residue analysis of pesticides by GC/ECD, 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) The mean values of heavy metal contents (mg/kg) for the samples were as follows: Yuldahanso-tang (before decoction - Pb; 1.85, Cd; 0.148, As; 0.042 and Hg; 0.003, after decoction - Pb; 0.096, Cd; 0.006, As; 0.006 and Hg; 0.002), Chongsimyonja-tang (before decoction - Pb; 1.193, Cd; 0.094, As; 0.084 and Hg; 0.008, after decoction - Pb; 0.053, Cd; 0.007, As; 0.011 and Hg; not detected) and Taeyeumjowee-tang (before decoction - Pb; 0.878, Cd; 0.078, As; 0.302 and Hg; 0.004, after decoction - Pb; 0.079, Cd; 0.005, As; 0.006 and Hg; not dectcted). 2) Contents (mg/kg) of residual pesticides before/after decoction in all samples were not detected. 3) Contents (mg/kg) of sulfur dioxide (SO2) before decoction in Yuldahanso-tang, Chongsimyonja-tang and Taeyeumjowee-tang exhibited 6.1, 37.8, 31.5 and 19.7, respectively. However, contents of sulfur dioxide after decoction in all samples were not detected. 4. Conclusion These results will be used to establish a criterion of heavy metals, residual pesticides and sulfur dioxide.

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.

HPLC-PDA를 이용한 오약순기산 중 6종 성분의 동시분석

서창섭,김정훈,신현규,Seo, Chang-Seob,Kim, Jung-Hoon,Shin, Hyeun-Kyoo 대한한의학방제학회 2012 大韓韓醫學方劑學會誌 Vol.20 No.2

Objectives : Oyaksungi-san(Wuyaoshunqisan) has been used for treatment of stroke and rheumatoid arthritis in Korea. In this study, a simple and accurate high-performance liquid chromatography(HPLC) method was established for simultaneous determination of six main components, liquiritin, ferulic acid, naringin, hesperidin, neohesperidin, and glycyrrhizin in Oyaksungi-san, a traditional Korean herbal prescription. Methods : The analytical column for separation of six constituents was used a Gemini $C_{18}$ column maintained at $40^{\circ}C$. The mobile phase consisted of two solvent systems, 1.0% (v/v) acetic acid in $H_2O$ (A) and 1.0% (v/v) acetic acid in acetonitrile (B) by gradient flow. The flow rate was 1.0 mL/min and the detector was a photodiode array (PDA) set at 254 nm for glycyrrhizin, 280 nm for liquiritin, naringin, hesperidin, and neohesperidin, and 320 nm for ferulic acid. Results : Calibration curves were acquired with $r^2$ values ${\geq}0.9998$. The results of recovery test were 91.58%-105.90% with a relative standard deviations (RSDs, %) value less than 2.0%. The values of RSD for intra- and inter-day precision were 0.03%-1.72% and 0.03%-1.63%, respectively. The contents of the six compounds in Oyaksungi-san were 0.33-9.30 mg/g. Conclusions : The newly established HPLC method will be helpful to improve quality control of Oyaksungi-san.

평위산 전탕팩의 장기보존 시험에 따른 유통기한 설정

서창섭,김정훈,임순희,신현규,Seo, Chang-Seob,Kim, Jung-Hoon,Lim, Soon-Hee,Shin, Hyeun-Kyoo 대한한의학방제학회 2011 大韓韓醫學方劑學會誌 Vol.19 No.1

Objectives : To estimate the shelf-life by long-term storage test of Pyungwi-san. Methods : Experiments were conducted to evaluate the stability such as the selected physicochemical, heavy metal, microbilogical experiment under an acceleration test and long-term storage test of Pyungwi-san in different storage under room temperature, refrigeration and freezing. Futhermore, HPLC analysis was performed for the determinations of glycyrrhizin in the Pyungwi-san on an Inertsil ODS-3 column(250 mm ${\times}$ 4.6 mm, 5 um) using solvent 35% acetonitrile include 0.05% phosphoric acid at 254 nm. The flow rate was 1.0 mL/min. Results : The significant change was not showed in pH, heavy metal, microbiological, identification test and quantitative analysis based on acceleration test and long-term storage test. Retention time of glycyrrhizin in HPLC chromatogram was about 16.065 min and calibration curve showed good linearity($R^2$ = 0.9999). The contents of glycyrrhizin in acceleration test and long-term storage test were 0.068~0.076 mg/mL and 0.066~0.077 mg/mL, respectively. Shelf-lifes of room temperature, refrigeration and freezing by long-term storage test were predicted 41, 24 and 34 months, respectively. Conclusions : The suggested shelf-life would be helpful on the storage and distribution of herbal medicine.

한방처방의 전탕 전과 후의 위해물질 농도변화 -보양.보음 처방을 중심으로 -

서창섭,황대선,이준경,하혜경,천진미,엄영란,장설,신현규,Seo, Chang-Seob,Huang, Dae-Sun,Lee, Jun-Kyoung,Ha, Hye-Kyoung,Chun, Jin-Mi,Um, Young-Ran,Jang, Seol,Shin, Hyeun-Kyoo 대한한의학방제학회 2009 大韓韓醫學方劑學會誌 Vol.17 No.2

Objective : To compare the contents of heavy metals, residual pesticides and sulfur dioxide before/after a decoction. Methods : The heavy metal contents before/after a decoction were measured by Inductively Coupled Plasma Atomic Emission Spectrometer (ICP-AES) and mercury analyzer. In order to analyze pesticides in 4 samples 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 ($SO_2$) were performed by Monier-Williams distillation method. Results: 1. The mean values of heavy metal contents (mg/kg) for the samples were as follows: Jaeumganghwa-tang (before decoction - Pb; 1.190, Cd; 0.184, As; 0.099 and Hg; 0.028, after decoction - Pb; .033, Cd; 0.003, As; 0.005 and Hg; 0.001), Yukmijiwhang-tang (before decoction - Pb; 0.484, Cd; 0.133, As; 0.053 and Hg; 0.009, after decoction - Pb; 0.065, Cd; 0.008, As; 0.007 and Hg; not detected), Bojungikgi-tang (before decoction - Pb; 0.863, Cd; 0.197, As; below 0.016 and Hg; 0.011, after decoction - Pb; 0.071, Cd; 0.009, As; 0.004 and Hg; 0.001) and Ssangwha-tang (before decoction - Pb; 1.511, Cd; 0.212, As; 0.094 and Hg; 0.016, after decoction - Pb; 0.029, Cd; 0.006, As; 0.005 and Hg; 0.0004). 2. Contents (mg/kg) of sulfur dioxide ($SO_2$) before a decoction in Jaeumganghwa-tang, Yukmijiwhang-tang and Ssangwha-tang exhibited 22.7, 107.3 and 5.5, respectively. However, contents of sulfur dioxide after a decoction in all samples were not detected. 3. Contents (mg/kg) of residual pesticides before/after a decoction in all samples were not detected. Conclusion : These results will be used to establish a criterion of heavy metals, residual pesticides and sulfur dioxide.

방풍통성산의 동시분석 및 항산화 효능 연구

서창섭,김온순,신현규,Seo, Chang-Seob,Kim, Ohn Soon,Shin, Hyeun-Kyoo 대한한의학방제학회 2013 大韓韓醫學方劑學會誌 Vol.21 No.2

Objectives : We carry out the simultaneous quantification for quality control of four components in Bangpungtongseong-san (BPTSS) sample. In addition, we assessed the antioxidant effects of BPTSS sample. Methods : The used column for separation and analysis of four compounds was Luna C18 column and column oven temperature was maintained at $40^{\circ}C$. The mobile phase for simultaneous determination consisted of two solvent systems, 1.0% acetic acid in water and 1.0% acetic acid in acetonitrile. High performance liquid chromatography-photodiode array (HPLC-PDA) method for analysis was performed at a flow rate of 1.0 mL/min with PDA detection at 254 and 280 nm. The injection volume was 10 ${\mu}L$. The antioxidant activities of BPTSS were evaluated 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 inhibitory effects on low-density lipoprotein (LDL) oxidation were evaluated by the formation of thiobarbituric acid relative substances (TBARS) and relative electrophoretic mobility (REM). Results : Calibration curves were acquired with $r^2{\geq}0.9999$. The values of limit of detection (LOD) and quantification (LOQ) were 0.06-0.29 ${\mu}g/mL$ and 0.20-0.98 ${\mu}g/mL$, respectively. The amounts of geniposide, liquiritin, baicalin, and glycyrrhizin in BPTSS were 5.06, 7.33, 27.56, and 7.81 mg/g, respectively. The BPTSS showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction (RC50) against ABTS and DPPH radicals were 72.51 ${\mu}g/mL$ and 128.49 ${\mu}g/mL$. Furthermore, GMGHT reduced the oxidation properties of LDL induced by CuSO4. Conclusions : The established HPLC-PDA method will be helpful to improve quality control of BPTSS. In addition, BPTSS has potentials as therapeutic agent on anti-atherosclerosis.

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.

LC-MS/MS를 이용한 이중탕(理中湯)의 정량분석 및 항산화 활성

서창섭,김온순,김예지,신현규,Seo, Chang-Seob,Kim, Ohn Soon,Kim, Yeji,Shin, Hyeun-Kyoo 대한한의학방제학회 2013 大韓韓醫學方劑學會誌 Vol.21 No.1

Objectives : Leejung-tang (Lizhong-tang) has been used for treatment of gastrointestinal disorders in Korea. In this study, we performed quantification analysis of five marker components, liquiritin, ginsenoside Rb1, ginsenoside Rg1, glycyrrhizin, and 6-gingerol in Leejung-tang using a ultra performance liquid chromatography- electrospray ionization-mass spectrometer (UPLC-ESI-MS). In addition, we evaluated antioxidant activity of Leejung- tang. Methods : The column for separation of five constituents used a UPLC BEH C18 ($100{\times}2.1mm$, $1.7{\mu}m$) maintained at $45^{\circ}C$. The mobile phase consisted of two solvent systems, 0.1% (v/v) formic acid in H2O (A) and CH3CN (B) by gradient flow. The flow rate was 0.3 mL/min with detection at mass spectrometer. The antioxidative activities conduct an experiment on 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities of Leejung-tang. Results : Calibration curves of five marker compounds were acquired with r2 values > 0.99. The amount of the five compounds in Leejung-tang were 0.07 - 0.84 mg/g. The concentration required for 50% reduction (RC50) against ABTS radical was 119.02 ug/mL. In addition, the scavenging against DPPH radical of Leejung-tang was 11.4%, 14.5%, 19.8%, 29.6%, and 49.2% at 25 ug/mL, $50{\mu}g/mL$, $100{\mu}g/mL$, $200{\mu}g/mL$, and $400{\mu}g/mL$, respectively. Conclusions : The established LC-MS/MS method will be helpful to improve quality control of Leejung-tang. In addition, Leejung-tang is a potential antioxidant therapeutic agent.

태음인(太陰人) 청심연자탕(淸心蓮子湯)의 동시분석 및 항비만 효과

서창섭,정수진,김정훈,유새롬,신현규,Seo, Chang-Seob,Jeong, Soo-Jin,Kim, Jung-Hoon,Yoo, Sae-Rom,Shin, Hyeun-Kyoo 사상체질의학회 2013 사상체질의학회지 Vol.25 No.1

Objectives We performed the simultaneous analysis for three compounds in Cheongsimyeonja-tang (CSYJT) and evaluated its anti-obesity effect. Methods The column for separation of three compounds was used Gemini $C_{18}$ column and maintained at $40^{\circ}C$. The mobile phase for gradient elution consisted of two solvent systems. The analysis was carried out at a flow rate of 1.0 mL/min with PDA detection at 275 nm. The injection volume was $10{\mu}L$. 3T3-L1 preadipocytes were differentiated into adipocytes by adding insulin, dexamethasone and 3-isobutyl-1-methylxanthine (IBMX) for 8 days in the absence or presence of CSYJT. Anti-obesity effects of CSYJT were evaluated by Oil Red O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity, triglyceride contents, and leptin production. Results Calibration curves were acquired with $r^2$ >0.9999. The contents of baicalin, wogonoside and baicalein in CSYJT were 14.54-14.65 mg/g, 5.24-5.27 mg/g and 0.01-0.02 mg/g, respectively. CSYJT showed inhibitory effect on lipid accumulation and GPDH activity in the differentiated 3T3-L1 cells. Furthermore, CSYJT significantly decreased contents of triglyceride and leptin production in 3T3-L1 adipocytes. Conclusions These results will be helpful to improve quality control and anti-obesity effect of Taeeumin CSYJT.