• Journal of Ginseng Research
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• 제42권2호
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• pp.149-157
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• 2018

Background: Panax notoginseng leaves (PNL) exhibit extensive activities, but few analytical methods have been established to exclusively determine the dammarane triterpene saponins in PNL. Methods: Ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC/Q-TOF MS) and HPLC-UV methods were developed for the qualitative and quantitative analysis of ginsenosides in PNL, respectively. Results: Extraction conditions, including solvents and extraction methods, were optimized, which showed that ginsenosides Rc and Rb3, the main components of PNL, are transformed to notoginsenosides Fe and Fd, respectively, in the presence of water, by removing a glucose residue from position C-3 via possible enzymatic hydrolysis. A total of 57 saponins were identified in the methanolic extract of PNL by UPLC/Q-TOF MS. Among them, 19 components were unambiguously characterized by their reference substances. Additionally, seven saponins of PNL-ginsenosides Rb1, Rc, Rb2, and Rb3, and notoginsenosides Fc, Fe, and Fd-were quantified using the HPLC-UV method after extraction with methanol. The separation of analytes, particularly the separation of notoginsenoside Fc and ginsenoside Rc, was achieved on a Zorbax ODS C8 column at a temperature of $35^{\circ}C$. This developed HPLC-UV method provides an adequate linearity ($r^2$ > 0.999), repeatability (relative standard deviation, RSD < 2.98%), and inter- and intraday variations (RSD < 4.40%) with recovery (98.7-106.1%) of seven saponins concerned. This validated method was also conducted to determine seven components in 10 batches of PNL. Conclusion: These findings are beneficial to the quality control of PNL and its relevant products.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2006년도 추계학술대회
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• pp.65-74
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• 2006

Modem drug discovery requires rapid pharmacokinetic evaluation of chemically diverse compounds for early candidate selection. This demands the development of analytical methods that offer high-throughput of samples. Naturally, liquid chromatography / tandem mass spectrometry (LC-MS/MS) is choice of the analytical method because of its superior sensitivity and selectivity. As a result of the short analysis time(typically 3-5min) by LC-MS/MS, sample preparation has become the rate- determining step in the whole analytical cycle. Consequently tremendous efforts are being made to speed up and automate this step. In a typical automated 96-well SPE(solid-phase extraction) procedure, plasma samples are transferred to the 96-well SPE plate, internal standard and aqueous buffer solutions are added and then vacuum is applied using the robotic liquid handling system. It takes only 20-90 min to process 96 samples by automated SPE and the analyst is physically occupied for only approximately 10 min. Recently, the ultra-high flow rate liquid chromatography (turbulent-flow chromatography)has sparked a huge interest for rapid and direct quantitation of drugs in plasma. There is no sample preparation except for sample aliquotting, internal standard addition and centrifugation. This type of analysis is achieved by using a small diameter column with a large particle size(30-5O ${\mu}$m) and a high flow rate, typically between 3-5 ml/min. Silica-based monolithic HPLC columns contain a novel chromatographic support in which the traditional particulate packing has been replaced with a single, continuous network (monolith) of pcrous silica. The main advantage of such a network is decreased backpressure due to macropores (2 ${\mu}$m) throughout the network. This allows high flow rates, and hence fast analyses that are unattainable with traditional particulate columns. The reduction of particle diameter in HPLC results in increased column efficiency. use of small particles (<2 urn), however, requires p.essu.es beyond the traditional 6,000 psi of conventional pumping devices. Instrumental development in recent years has resulted in pumping devices capable of handling the requirements of columns packed with small particles. The staggered parallel HPLC system consists of four fully independent binary HPLC pumps, a modified auto sampler, and a series of switching and selector valves all controlled by a single computer program. The system improves sample throughput without sacrificing chromatographic separation or data quality. Sample throughput can be increased nearly four-fold without requiring significant changes in current analytical procedures. The process of Bioanalytical Method Validation is required by the FDA to assess and verify the performance of a chronlatographic method prior to its application in sample analysis. The validation should address the selectivity, linearity, accuracy, precision and stability of the method. This presentation will provide all overview of the work required to accomplish a full validation and show how a chromatographic method is suitable for toxirokinetic sample analysis. A liquid chromatography/tandem mass spectrometry (LC-MS/MS) method developed to quantitate drug levels in dog plasma will be used as an example of tile process.

• 한국환경보건학회지
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• 제41권5호
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• pp.289-298
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• 2015

Objectives: The method of analyzing urinary arsenic by flow injection hydride generation atomic absorption spectrometry (FI-HG-AAS) is generally used because it shows relatively greater sensitivity, low detection limits, low blocking action, and is simple to operate. In this study, the results of analysis according to three pre-reductants commonly used in the FI-HG-AAS method were compared with each other. Methods: To analyze urinary arsenic, nineteen urine samples were collected from adults aged 43-79 years old without occupational arsenic exposure. Analysis equipment was FI-HG-AAS (AAnalyst 800/FIAS 400, Perkin- Elmer Inc., USA). The three pre-reductants were potassium iodide (KI/AA), C3H7NO2S (L-cysteine), and a mixture of KI/AA and L-cysteine (KI/AA&L-cysteine). Results: In the results of the analysis, the recovery rate of the method using KI/AA was 82.3%, 95.7% for Lcysteine, and 123.5% for KI/AA and L-cysteine combined. When compared with the results by use of high performance liquid chromatography inductively-coupled plasma mass spectrometry (HPLC-ICP-MS), the method using L-cysteine was the closest to those using HPLC-ICP-MS ($98.57{\mu}g/L$ for HPLC-ICP-MS; $74.96{\mu}g/L$ for L-cysteine; $69.23{\mu}g/L$ for KI/AA and L-cysteine; $13.06{\mu}g/L$ for KI/AA) and were significantly correlated (R2=0.882). In addition, they showed the lowest coefficient of variation in the results between two laboratories that applied the same method. Conclusion: The efficiency of hydride generation is considered highly important to the analysis of urinary arsenic via FI-HG-AAS. This study suggests that using L-cysteine as a pre-reductant may be suitable and the most rational among the FI-Hg-AAS methods using pre-reductants.

• 대한화학회지
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• 제46권6호
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• pp.535-540
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• 2002

남조류 독소인 마이크로시틴은 호수에서 매우 미량 존재하기 때문에 이를 정확히 정량 분석하기가 쉽지 않다. 본 연구에서는 소양호에서 채취한 물시료 중에 포함된 남조류 독소, 마이크로시틴을 정량 분석하기 위하여 두 가지 서로 다른 정량 분석법을 시도하였으며 이 둘의 정량 결과를 비교${\cdot}$분석 하였다. 첫째는 고체상 추출과 HPLC(High Performance Liquid Chromatography)를 이용한 분석법을 이용하였으며 둘째는 마이크로시틴의 단일클론항체를 이용한 효소면역흡착분석법(Enzyme-Linked Immunosorbent Assay,ELISA)을 이용하였다.

• 분석과학
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• 제25권6호
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• pp.388-394
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• 2012

남조류 독소인 마이크로시스틴은 여름철 우리나라 여러 호수들에 존재하여 물고기와 가축 그리고 인간에게 강한 독성을 나타내는 독소이다. 지금까지는 남조류독소인 마이크로시스틴을 분석하기 위하여 HPLC를 사용하거나 ELISA 분석법을 이용하였으나 본 연구에서는 대량배양을 통해서 얻어진 남조류들 속에 미량존재하는 마이크로시스틴을 분석하기 위해 스트립을 이용하는 새로운 분석방법을 시도하였다. 이 분석법은 항원-항체기술을 이용하여 높은 특이도(specifisity)와 테스트 스트립(test strip)의 신속성, 그리고, 형광리더(reader)의 고감도(sensitivity) 분석능력 등의 장점을 이용하였다. 따라서 새로 개발된 분석법을 이용하면 다양한 물시료속에 존재하는 마이크로시스틴을 독소 표준품 없이 단시간에 고감도로 분석할 수 있다.

• 대한한방내과학회지
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• 제27권1호
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• pp.55-71
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• 2006

Objectives : To find out more pharmacologically efficient way of extraction of herbal dicoction, Gamichungsangbohatang (GMCSBHT). Methods : Several main components of GMCSBHT was compared and analysed between hot water extract of GMCSBHT and Alcohol(70% ethanol) extract of GMCSBHT via HPLC method. Results : Hot water extract of GMCSBHT showed relatively more component content than ethanol extract of GMCSBHT. Also weighted mean of main components of hot water extraction of GMCSBHT was higher than that of alcohol extract of GMCSBHT. But from chromatographic pattern analysis of matching ratio and similarity ratio showed that these two forms of extracts might have different chemical composition, and 3D PDA plot of alcohol extract of GMCSBHT showed high peaks near UV $190{\sim}220nm$ which was invisible in hot water extract of GMCSBHT. Conclusion : Alcohol extract of GMCSBHT may have some special components which do not exist in hot water extract of GMCSBHT.

• 한국치위생학회지
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• 제17권3호
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• pp.381-394
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• 2017

Objectives: This study aims to evaluate carbonated drinks induced dental caries with qualitative analysis and to compare with oral organic acids including lactate, acetate, propionate, formate, butyrate, pyruvate and valerate which cause caries when taking either 10% sucrose drinks or carbonated drinks. Methods: Saliva was collected from six study subjects before and after (start, 5, 10, 30 minutes) taking water intake upon (A) 10% sucrose intake, (B) 10% sucrose intake, and (C) carbonated drink intake, then they were centrifuged at 1,200 rpm followed by removing bacteria and enzymes with syringe filtering, performing a qualitative analysis with HPLC conductivity detection (GP50 gradient pump, ED 50 detector) after saliva pre-treatment under isocratic 100 mM NaOH mobile phase. Results: Higher risk of dental caries was evaluated in order of C>B>A, with the results of total oral organic acids' concentration, lactates of organic acids and organic acids produced after 5 minutes from the 3 types of drinks intake. Conclusions: Carbonated beverages were estimated to develop higher dental caries induction than beverages containing 10% sucrose because of the high organic acid concentration in the mouth after its intake.

• 분석과학
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• 제5권2호
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• pp.153-158
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• 1992

본 연구에서는 이동상 성질이 GC와 HPLC와는 전혀 다른 초임계 유체 크로마토그래피(SFC)를 사용함으로써 종래의 크로마토그래피의 방법으로는 분석하기 힘든 물질을 분석하는 방법을 개발하였다. GC로는 분리하기 힘든 긴 고리를 가진 Hydrocarbons이나 Mink Oils, Soybean oils 등이 SFC로 잘 분리될 수 있었다. 그 이유는 SFC에서는 시료가 갖고 있는 낮은 기화성이나 열에 불안정한 점 등이 이동상을 초임계 상태의 이산화탄소를 사용함으로써 극복되어질 수 있었다. 이 연구에서는 한걸음 더 나아가 극성이 강한 지방산 및 농약류를 분석하는 방법을 개발함으로써 SFC의 가장 큰 약점인 극성이 있는 시료의 처리 문제를 극복하고자 시도하였다.

• Journal of Ginseng Research
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• 제46권3호
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• pp.489-495
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• 2022

Background: Although ginsenosides and saponins in Korea red ginseng (KRG) shows various pharmacological roles, their roles in the inflammatory response are little known. This study investigated the anti-inflammatory role of ginsenosides identified from KRG saponin fraction (RGSF) and the potential mechanism in macrophages. Methods: The ginsenoside composition of RGSF was identified by high-performance liquid chromatography (HPLC) analysis. An anti-inflammatory effect of RGSF and its mechanisms were studied using nitric oxide (NO) and prostaglandin E₂ (PGE₂) production assays, mRNA expression analyses of inflammatory genes and cytokines, luciferase reporter gene assays of transcription factors, and Western blot analyses of inflammatory signaling pathways using the lipopolysaccharide (LPS)-treated RAW264.7 cells. Results: HPLC analysis identified the types and amounts of various panaxadiol ginsenosides in RGSF. RGSF reduced the generation of inflammatory molecules and mRNA levels of inflammatory enzymes and cytokines in LPS-treated RAW264.7 cells. Additionally, RGSF inhibited the signaling pathways of NF-κB and AP-1 by suppressing both transcriptional factors and signaling molecules in LPS-treated RAW264.7 cells. Conclusion: RGSF contains ginsenosides that have anti-inflammatory action via restraining the NF-κB and AP-1 signaling pathways in macrophages during inflammatory responses.

• 한국작물학회지
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• 제37권4호
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• pp.377-382
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• 1992

본 연구는 고항산화 양질 육종의 체계확립을 위해서 참깨에 있는 특수성분인 sesamin, sesamolin의 표준물질을 동정하고 그 분석기술을 개발하였는바 그 결과는 다음과 같다. 1. 한국산 참깨로부터 sesamin, sesamolin 성분을 분리하여 화학적 구조를 분광학적 방법(UV, $^1$H와 $^{13}$C-NMR, MS)으로 확인하여 sesamin, sesamolin의 표준물질을 동정하였다. 2. 참깨 lignan성분의 추출 및 분리에는 Unsaponifiable fration 보다 MeOH 냉온처리가 더 효과적이었다. 3. 검량선의 회귀직선의 방정식은 sesamin에서 y=53924.43＋1277.93x(r=0.9993)이었고 sesamolin의 경우는 y=11034.95＋1188.38x(r=0.9994)로 추정되었다. 4. 한국산 참깨의 lignan 성분함량은 단백깨의 경우 sesamin 0.42% sesamolin 0.3%로 나타났다.