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.

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.

산조인의 수치에 따른 Spinosin 함량 비교

서창섭,김병수,김정훈,신현규 한국생약학회 2016 생약학회지 Vol.47 No.4

The aim of this study was to compare the amount of spinosin in the 70% ethanol extracts of non-processed Zizyphi Semem (ZS) and processed ZS by roasting using a high-performance liquid chromatography equipped with photodiode array detector. Separation of the spinosin was used SunFireTM C18 analytical column (5 mm, 4.6×150 mm) using two mobile phase consisting of distilled water and acetonitrile, both with 1.0% (v/v) acetic acid. The flow rate was 1.0 mL/min and injection volume was 10 mL. Calibration curve of the spinosin was y = 22339.45x+483.99 in tested concentration range (1.28-20.00 mg/mL) and correlation coefficient was 1.0000. In non-processed ZS sample, the amount of the spinosin was 0.94 m/g, while, the amount of the marker compound in processed ZS samples were 0.66-1.10 mg/g.

LC-MS/MS를 이용한 향소산 중 15종 성분의 정량분석

서창섭,신현규 한국생약학회 2016 생약학회지 Vol.47 No.4

Hyangso-san is a traditional herbal medicine that consists of the seven herbal medicines, Cyperi Rhizoma, Perillae Folium, Atractylodis Rhizoma, Citri Unshius Pericarpium, Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma Crudus, and Allii Fistulosi Bulbus. Hyangso-san has long been clinically used to treat the influenza, including headache, ferver, chills, and pantalgia. In this study, we were performed the simultaneous analysis of the 15 marker compounds (liquiritin apioside, liquiritin, ferulic acid, naringin, hesperidin, rosmarinic acid, liquiritigenin, kaempferol, glycyrrhizin, nobiletin, 6-gingerol, elemicin, atractylenolide III, nootkatone, and atractylenolide I) in Hyangso-san using liquid chromatography–electrospray ionization-mass spectrometry (LC-ESI-MS/MS). Column for the separation of the 15 ingredients was used a Waters Acquity UPLC BEH C18 analytical column (2.1×100 mm, 1.7 mm) at 45oC by using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient condition. Identifications of all analytes were performed using a Waters ACQUITY TQD LC-MS/MS system. The flow rate and injection volume were 0.3 mL/min and 2.0 mL, respectively. Correlation coefficient of the calibration curve was ³0.9958. The values of limits of detection and quantification of the 15 components were 0.002-4.29 and 0.01-12.88 ng/mL, respectively. The result of an analysis using the established LC-MS/MS method, kaempferol and atractylenolide I were not detected, while other 13 compounds were 0.08-56.87 mg/g in lyophilized Hyangso-san sample.

HPLC-PDA를 이용한 보중익기탕 중 Liquiritin, Nodakenin, Hesperidin 및 Glycyrrhizin의 동시분석

서창섭,김정훈,신현규 대한약학회 2013 약학회지 Vol.57 No.3

Bojungikgi-tang has been widely used for enhancement of physical fitness in Korea. The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of four marker compounds, liquiritin, nodakenin, hesperidin, and glycyrrhizin in Bojungikgi-tang (Buzhongyiqi-tang in Chinese, Hochuekkito in Japanese), a traditional Korean herbal prescription. The column for optimizing HPLC separation was used a Gemini C18 column at column oven temperature of 40 o C with 1.0% (v/v) aqueous acetic acid (A) and 1.0% (v/v) acetic acid in acetonitirle (B) by gradient flow. The flow rate was 1.0 ml/min and the detector was a photodiode array (PDA) set at 254 nm, 280 nm, and 335 nm. Calibration curves of four components were acquired with r 2 values ≥0.9999. The recoveries were found to range 92.11~105.68% with relative standard deviations (RSDs, %) value less than 2.50%. The RSD values of intra- and inter-day precision were 0.07~2.50% and 0.16~1.99%, respectively. The contents of liquiritin, nodakenin, hesperidin and glycyrrhizin in Bojungikgi-tang were 3.85~3.92 mg/g, 2.27~2.32 mg/g, 4.14~4.19 mg/g, and 3.39~3.45 mg/g, respec- tively. The established simultaneous analysis method will be effective for quality control of Bojungikgi-tang.

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 반하사심탕 물 추출물 중 13종 성분의 함량분석

서창섭,신현규 한국생약학회 2016 생약학회지 Vol.47 No.1

Banhasasim-tang is a well-known traditional Korean herbal formula and has been used clinically for the treatment of gastric disease, including acute and chronic gastritis, diarrhea and gastric ulcers in Korea. In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometer method was developed for the quantitative determination of the 13 marker constituents, homogentisic acid (1), 3,4-dihydroxybenzaldehyde (2), spinosin (3), liquiritin (4), baicalin (5), ginsenoside Rg1 (6), liquiritigenin (7), wogonoside (8), ginsenoside Rb1 (9), baicalein (10), glycyrrhizin (11), wogonin (12), and 6-gingerol (13) in Banhasasim-tang decoction. Separation of the compounds 1-13 was using an UPLC BEH C18 (100×2.1 mm, 1.7 mm) column and column oven temperature was maintained at 45oC. The mobile phase consisted of 0.1% (v/v) formic acid in water (A) and acetonitrile (B) by gradient elution. The injection volume and flow rate were 2.0 mL and 0.3 mL/min, respectively. Calibration curves of the compounds 1-13 were showed with r2 values ³ 0.9908. The limit of detection and limit of quantification values of the compounds 1-13 were 0.04-1.11 ng/mL and 0.13-3.33 ng/mL, respectively. Among the these compounds, the compounds 1-3 were not detected, while the compounds 4-13 were detected in the ranges of 3.20-107,062.98 mg/g in Banhasasim-tang sample.

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.

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.

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.