• 약학회지
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• 제58권5호
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• pp.314-321
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• 2014

Cibotii Rhizoma, the dried rhizome of Cibotium barometz J. Smith (C. barometz), has long been used to treat bone or nervous system disorders. In this regard, we isolated three main phenolic compounds, onitin-4-O-${\beta}$-D-glucopyranoside (1), irisdichototins E & F epimeric mixture (2), and protocatechuic acid (3) from C. barometz methanol extract. In addition, we screened their antioxidative activities by DPPH, ABTS radical, and superoxide scavenging assays. Among these three compounds, irisdichototins E & F and protocatechuic acid showed strong antioxidant activities. Also, the antioxidant activities of the C. barometz extracts were proportional to the contents of irisdichototins E & F and protocatechuic acid, thus these two phenolic compounds could be main active compounds of C. barometz. In addition, onitin-4-O-${\beta}$-D-glucopyranoside is considered as a marker compound of C. barometz because this compound is specifically contained in C. barometz which belongs to Pteridophyta order. A rapid analysis method for the simultaneous determination of phenolic compounds was also developed by UPLC (Ultra Performance Liquid Chromatography). Using the developed method, the two active compounds (irisdichototins E & F and protocatechuic acid) and a marker compound (onitin-4-O-${\beta}$-D-glucopyranoside) were successfully quantified in 14 commercial samples that were collected from different regions.

• Natural Product Sciences
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• 제22권2호
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• pp.93-101
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• 2016

For efficient quality control of the Samryeongbaekchul-san decoction, a powerful and accurate an ultra-performance liquid chromatography (UPLC) coupled with electrospray ionization (ESI) tandem mass spectrometry (MS) method was developed for quantitative analysis of the thirteen constituents: allantoin (1), spinosin (2), liquiritin (3), ginsenoside Rg1 (4), liquiritigenin (5), platycodin D2 (6), platycodin D (7), ginsenoside Rb1 (8), glycyrrhizin (9), 6-gingerol (10), atractylenolide III (11), atractylenolide II (12), and atractylenolide I (13). Separation of the compounds 1 - 13 was performed on a UPLC BEH $C_{18}$ column ($2.1{\times}100mm$, $1.7{\mu}m$) at a column temperature of $40^{\circ}C$ with a gradient solvent system of 0.1% (v/v) formic acid aqueous-acetonitrile. The flow rate and injection volume were 0.3 mL/min and $2.0{\mu}L$. Calibration curves of all compounds were showed good linearity with values of the correlation coefficient ${\geq}0.9920$ within the test ranges. The values of limits of detection and quantification for all analytes were 0.04 - 4.53 ng/mL and 0.13 - 13.60 ng/mL. The result of an experiment, compounds 2, 6, 12, and 13 were not detected while compounds 1, 3 - 5, and 7 - 11 were detected with 1,570.42, 5,239.85, 299.35, 318.88, 562.27, 340.87, 12,253.69, 73.80, and $115.01{\mu}g/g$, respectively.

• 생약학회지
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• 제45권4호
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• pp.338-345
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• 2014

HPL-4 is a new herbal formulation developed for the treatment of osteoarthritis. In this study, we took HPL-4 to develop a method for simultaneous determination of nine marker compounds (protocatechuic acid, chlorogenic acid, liriodendrin, nodakenin, ${\beta}$-$\small{D}$-(3-O-sinapoyl)frucofuranosyl-${\alpha}$-$\small{D}$-(6-O-sinapoyl)glucopyranoside, quercitrin, 6-gingerol, decursin and decursinol angelate) present in the formulation. The analytes were separated by UPLC system consisting of diode array detector at 205 nm and RP-amide column, and solvent system of $ACN/0.1%H_3PO_4$. Analytical method was validated to evaluate its linearity, limit of detection (LOD), limit of quantification (LOQ), precision, and accuracy. All standards showed good linearity ($R^2$ >0.99) in the rage of $0.25-400.0{\mu}g/mL$. The LOD and LOQ were within the range of 0.021-0.148 and $0.070-0.448{\mu}g/mL$, respectively. Intra- and inter-day precision was less than RSD 4.0% and the accuracy was range from 92.00-104.81% with RSD<4.2%. The results suggest that the developed UPLC method is precise, accurate and effective, and could be applied for quality control of HPL-4 formulation.

• 분석과학
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• 제33권3호
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• pp.125-133
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• 2020

Methylisothiazolinone (MIT) and chloromethylisothiazolinone (CMIT) cause allergic contact dermatitis and are banned cosmetics ingredients, except in rinse-off products. However, their presence has been detected in cosmetics. We report a UPLC-tandem MS/MS screening method for their simultaneous determination in cosmetics. To facilitate extraction from various matrices, pretreatment methods were developed for each sample type. The method was optimized through a series of assessments, including specificity, LOD, LOQ, linearity, recovery, stability, precision, and accuracy. The LODs and LOQs for MIT ranged from 0.054 and 0.163 ㎍ mL^-1 whereas those for CMIT ranged from 0.040 and 0.119 ㎍ mL^-1. The linear correlation coefficients (r²) were higher than 0.999. Relative standard deviations (RSDs) for both intra- and inter-day measurements ranged from 0.3 ~ 13.6 %. Recoveries at three different concentrations were within 87.9 ~ 118.9 %. The RSD for stability measurements of spiked samples was within 7 %. These results confirm the suitability of the developed method for the simultaneous quantitation of MIT and CMIT in cosmetics. Samples of 320 color cosmetics, including eyeshadows, solid lipsticks, liquid lipsticks, and nail polishes were analyzed using the developed method, and two of them were found to contain both MIT and CMIT and one of them was found to contain only MIT. This data and the method will aid the regulation of ingredients used in cosmetics.

• 대한본초학회지
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• 제26권2호
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• pp.11-15
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• 2011

Objectives : In the present study, the contents of cinnamic acid and cinnamaldehyde in three different parts of Cinnamomi Ramulus (CR) (the whole body, the bark part, and the wood part) was evaluated using UPLC (ultra performance liquid chromatography) in order to investigate a suitable cutting method. Methods : Analysis was performed on SMART LC with UV detector. Reference compounds were separated on Inertsil ODS-4 column ($2.1mm{\times}50mm$, $3{\mu}m$, GL Science, Japan) using isolation elution with water and acetonitrile each containing acetic acid at a flow rate of $500{\mu}L/min$. Additionally, samples of CR were purchased from pharmacy of medicinal herb. Results : The correlation coefficients of the cinnamic acid and cinnamaldehyde levels showed good linearity ($r^2{\geq}0.9999$) over the linear ranges. Furthermore, the bark part exhibited higher concentration levels of reference compounds than the wood part in all samples. In addition the bark exfoliation rates in oblique and perpendicular-long cut samples of CR were lower than the perpendicular-short cut samples. Conclusions : These results suggested that the optimal cutting method would be able to reduce the bark exfoliation. Therefore, the oblique or perpendicular-long cutting method is considered to be a better cutting type than the perpendicular-short cutting method.

• 한국양봉학회지
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• 제32권3호
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• pp.253-259
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• 2017

충치발생의 주요 원인균으로 알려진 S. mutans를 이용하여 건강기능식품으로 사용되는 프로폴리스를 9개 지역으로부터 수집한 후 생리활성 물질 탐색과 주요성분 함량에 따라 활성차이를 조사하였다. S. mutans에 대한 항균활성은 제주도, 경상북도, 경상남도, 충청북도, 충청남도, 전라북도, 전라남도, 경기도, 강원도 프로폴리스 및 pinocembrin에서 모두 관찰되었다. 제주도를 제외한 모든 프로폴리스에서 총 플라보노이드 함량이 10mg/g 이상으로 건강기능식품 기준 및 규격에 적합하였고, 19mg/g 이상일 경우 우수한 항균활성을 유사하게 나타내었다. 국산 프로폴리스의 주요성분으로 알려진 pinocembrin의 UPLC 정량분석에서는 제주도산 프로폴리스를 제외한 모든 시료에서 주성분으로 검출되었고 12mg/g 이상일 경우 항균활성에 큰 영향을 주는 것으로 확인되었다.

• Journal of Ginseng Research
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• 제37권3호
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• pp.341-348
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• 2013

Identification of the origins of Panax ginseng has been issued in Korea scientifically and economically. We describe a metabolomics approach used for discrimination and prediction of ginseng roots from different origins in Korea. The fresh ginseng roots from six ginseng cooperative associations (Gangwon, Gaeseong, Punggi, Chungbuk, Jeonbuk, and Anseong) were analyzed by UPLC-MS-based approach combined with orthogonal projections to latent structure-discriminant analysis multivariate analysis. The ginsengs from Gangwon and Gaeseong were easily differentiated. We further analyzed the metabolomics results in subgroups. Punggi, Chungbuk, Jeonbuk, and Anseong ginseng could be easily differentiated by the first two orthogonal components. As a validation of the discrimination model, we performed blind prediction tests of sample origins using an external test set. Our model predicted their geographical origins as 99.7% probability. The robust discriminatory power and statistical validity of our method suggest its general applicability for determining the origins of P. ginseng samples.

• 생약학회지
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• 제48권1호
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• pp.88-92
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• 2017

In the present study, we carried out quantitative analysis of chrysin and pinocembrin in Korean propolis by ultra performance liquid chromatography (UPLC) equipped with diode array detector. The separation was done using BEH C18 ($2.1{\times}50mm$, $1.7{\mu}m$) column with a mobile phase consisting of MeCN and 0.1% $H_3PO_4$ at 280 nm. The chromatographic method was validated for specificity, limit of detection, limit of quantification, linearity, precision, and accuracy. A quantitative analysis exhibited that the contents of the two compounds in Korean propolis collected from 8 inland areas except Jeju-do ranged from 3.1-46.0 mg/g. These results will be valuable as basic data for standardization of Korean propolis.

• 대한화장품학회지
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• 제33권4호
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• pp.263-267
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• 2007

미생물의 오염을 통해 화장품이 변질되거나 분해되는 것을 방지하여 소비자를 보호하기 위해 화장품에 보존제가 사용된다. 파라벤류는 제형화하기 쉽고, 활성 범위가 넓으며, pH에 화학적으로 안정하면서 저렴하여 거의 모든 종류의 화장품에 널리 사용된다. 페녹시에탄올과 클로페네신 역시 화장품에 일반적으로 사용되는 보존제로 보통 파라벤과 함께 사용된다. 파라벤의 독성은 일반적으로 낮지만, 손상된 피부에는 자극을 유발할 수 있으며, estrogenic 잠재성, 마취 효과 및 생식 독성의 가능성에 대한 논란이 있어 왔다. 따라서 파라벤은 배합한도 원료로 지정 관리되고 있으며, 페녹시에탄올과 클로페네신도 마찬가지로 최대 허용량이 지정되어 있다. 그러므로 제품 중 보존제의 함량을 관리하는 것은 중요하다. 그러나 일반적으로 사용되는 역상 액체크로마토그래프법으로는 이성질체를 포함한 6종의 파라벤과 페녹시에탄올 및 클로페네신을 동시에 분리 분석하기 어려웠다. 용출 시간이 길어져, 피크 모양이 나쁘고 분리능이 좋지 않아 정확한 정량이 불가능하였다. 본 연구에서는 ultra performance liquid $chromatography^{TM}\;(UPLC^{TM}$)를 이용하여 8종의 보존제를 10 min 이내에 동시분석을 시도하였다. 또한, International conference on harmonisation (ICH) 가이드라인의 밸리데이션 방법에 근거하여 본 시험법의 적합성을 검증하고, 로션, 팩트, 파운데이션 및 립글로스 등 다양한 제형에 적용이 가능함을 보였다. 본 시험법은 파라벤류를 포함한 다양한 보존제를 함유한 화장품 중 보존제의 함량을 단시간에 간편하고 정확하게 정량하는데 활용될 수 있을 것이다.360 nm)에서 3개의 피이크로 분리되었다. 분리된 3가지 성분은 luteolin, quercetin 및 kaempferol이었으며, 그들의 성분비는 각각 18.24 %, 58.79 %, 22.97 %로 quercetin의 함량이 가장 큰 것으로 나타났다. 루이보스 추출물의 ethylacetate 분획의 TLC 크로마토그램은 7개의 띠로 분리되었고, HPLE 크로마토그램은 9개의 피이크를 보여주었다. TLC와 HPLC의 띠와 피이크를 확인한 결과, HPLC의 9개의 피이크는 용리순서로 peak 1 (조성비 14.71 %)은 isoorientin, peak 2 (28.84 %)는 orientin peak 3 (5.63 %)은 vitexin, peak 4 (12.73 %)는 rutin과 isovitexin, peak 5 (9.24 %)는 hyperoside, peak 6 (5.40%)은 isoquercitrin, peak 7 (1.48 %)은 luteolin, peak 8 (17.61 %)은 quercetin 및 peak 9 (4.59 %)는 kaempferol로 확인되었다. Aglycone 분획은 elastase 저해활성($IC_{50}$)이 $9.08\;{\mu}g/mL$로 매우 큰 활성을 나타내었다. 이상의 결과들은 루이보스 추출물이 $^1O_2$ 혹은 다른 ROS를 소광시키거나 소거함으로써 그리고 ROS에 대항하여 세포막을 보호함으로써 생체계, 특히 태양 자외선에 노출된 피부에서 항산화제로서 작용할 수 있음을 가리키며, 루이보스 성분에 대한 분석과 ethylacetate 분획의 당 제거 실험 후 얻어진 aglycone 분획의 큰 elastase 저해활성으로부터

• 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.