• Title/Summary/Keyword: UPLC-UV

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Quantitative Analys is of Flavonoids in Hovenia dulcis by Region Using UPLC (UPLC를 이용한 지역별 헛개나무(Hovenia dulcis) 플라보노이드의 정량분석)

  • Dong Hwan Lee;Hyun-Jun Kim
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.100-100
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    • 2022
  • Hovenia dulcis is a herbal plant, which belongs to the Rhamnaceae family and is a native of Japan, China and Korea. Its fruit stalk is called 'Jiguja' in Korea. It has been traditionally used as a medicinal plant in East Asia. It was reported to have detoxification effects on alcohol poisoning, and antioxidant, antidiabetic etc. Sample of 5 g was extracted with 50 mL of 70% EtOH. The supernatant was filtered by 0.45 ㎛ membrane filter before analysis. The UPLC system was performed on Waters alliance UPLC HSS T3 column (2.1 × 100 mm, 1.7 ㎛) with a UV detector. The gradient system was a binary eluent of 0.1% formic acid in water(A) and 0.1% formic acid in acetonitrile(B) with gradient conditions as follows: Initial, 10% B; 1 min, 10% B; 4 min, 20% B; 10 min, 25% B; 12 min, 30% B; 14 min, 90% B; 17 min, 90% B; flow rate of 0.2 mL/min. The samples were injected by 2 µL and were detected at UV 355 nm. As a result of analysis, chromatographic patterns appeared in two cases: samples analyzed for ampelopsin and myricetin, and samples analyzed for taxifolin and quercetin. Among the four compounds, the largest regional difference was found to be taxifolin.

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Detection of phenobarbital adulteration in dietary supplements: simultaneous analysis of 16 sedative-hypnotics and sleep-inducers by ultra-high-performance liquid chromatography with UV detection (UPLC-UV) and quadruple Orbitrap mass spectrometry (Q-Orbitrap-MS)

  • Lee, Ji Hyun;Choi, Ji Yeon;Park, Hanna;Min, Ah Young;Kim, Nam Sook;Park, Seong Soo;Park, Sung-Kwan;Kang, Ho-il
    • Analytical Science and Technology
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    • v.32 no.1
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    • pp.24-34
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    • 2019
  • The safety of food is occasionally questionable, as there have been some reports of products contaminated with illegal adulterants. In this study, the presence of 16 sedative-hypnotics and sleep inducers in dietary supplements was determined by ultra-high-performance liquid chromatography with UV detection (UPLC-UV) and quadruple Orbitrap mass spectrometry (Q-Orbitrap-MS). The UPLC method was validated, providing a linearity (R2) of more than 0.999, and LODs and LOQs that ranged from 0.2 to 0.5 and 0.6 to $1.5{\mu}g\;mL^{-1}$, respectively. The repeatabilities were 0.2-8.4 % (intra-day) and 0.3-4.5 % (inter-day), and the accuracies were 89.0-117.0 % (intra-day) and 87.8-111.9 % (inter-day). The mean recoveries of the spiked samples ranged from 98.7 to 107.3 %. The relative standard deviation (%RSD) of the stability was less than 2.4 %. Using the developed method, one sedative-hypnotic compound, phenobarbital, was detected in one of the nineteen samples tested. In addition, the major characteristic fragment ions of each target compound were confirmed using Q-Orbitrap-MS for higher accuracy. Monitoring the presence of these 16 sedative-hypnotics and sleep inducers in dietary supplements should be pursued in the interest of human health, and the results of this study confirmed that the developed method has value for this application.

Qualitative and quantitative analysis of the saponins in Panax notoginseng leaves using ultra-performance liquid chromatography coupled with time-of-flight tandem mass spectrometry and high performance liquid chromatography coupled with UV detector

  • Liu, Fang;Ma, Ni;He, Chengwei;Hu, Yuanjia;Li, Peng;Chen, Meiwan;Su, Huanxing;Wan, Jian-Bo
    • Journal of Ginseng Research
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    • v.42 no.2
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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.

Development of Quantitative Analytical Method for Isoflavonoid Compounds from Fruits of Cudrania Tricuspidata (꾸지뽕나무 열매 isoflavonoid 화합물에 대한 정량분석법 개발)

  • Yoon, Sun Young;Kim, Seok Ju;Sim, Su Jin;Lee, Hak-Ju
    • Journal of the Korean Wood Science and Technology
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    • v.44 no.3
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    • pp.337-349
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    • 2016
  • In this study, an analytical method to evaluate the quality of isoflavonoid compounds purified and isolated from the fruits of Cudrania tricuspidata was developed and validated using Ultra Performance Liquid Chromatography (UPLC). The fruits of C. tricuspidata were extracted with methanol and further fractioned with n-hexane, ethyl acetate and water. The resulting ethyl acetate extract separated into four isoflavonoid compounds by a combination of silica gel and sephadex LH-20 column chromatography. The structures of the compounds were elucidated as alpinumisoflavone, 6,8-diprenyl genistein, 6,8-diprenyl orobol, 4'-O-methylalpinumisoflavone by various techniques such as UV-Vis, ESI-MS, $^1H\;NMR$ and $^{13}C\;NMR$ spectroscopy. Finally, a method to characterize the compounds was developed by using the UPLC equipped with a $C_{18}$ column and a gradient mobile phase system consisting of 2% acetic acid in water (solvent A) and 2% acetic acid in methanol (solvent B). The developed method was validated with the parameters such as selectivity, linearity, limit of detection, limit of quantitation, accuracy, and precision, which are defined by the ICH (International Conference on Harmonization). Using the validated method, the compounds in the fruits harvested in different months were also quantitatively analyzed. We propose this approach this approach can readily be utilized as an efficient evaluation method to quantify the extracts of C. tricuspidata.

Topical or oral treatment of peach flower extract attenuates UV-induced epidermal thickening, matrix metalloproteinase-13 expression and pro-inflammatory cytokine production in hairless mice skin

  • Kwak, Chung Shil;Yang, Jiwon;Shin, Chang-Yup;Chung, Jin Ho
    • Nutrition Research and Practice
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    • v.12 no.1
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    • pp.29-40
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    • 2018
  • BACKGROUND/OBJECTIVES: Ultraviolet radiation (UV) is a major cause of skin photoaging. Previous studies reported that ethanol extract (PET) of Prunus persica (L.) Batsch flowers (PPF, peach flowers) and its subfractions, particularly the ethylacetate (PEA) and n-butanol extracts (PBT), have potent antioxidant activity and attenuate the UV-induced matrix metalloproteinase (MMP) expression in human skin cells. In this study, we investigated the protective activity of PPF extract against UV-induced photoaging in a mouse model. MATERIALS/METHODS: Hairless mice were treated with PET or a mixture of PEA and PBT either topically or orally along with UV irradiation. Histological changes and biochemical alterations of mouse skin were examined. Major phenolic compounds in PPF extract were analyzed using an ACQUITY UPLC system. RESULTS: The overall effects of topical and oral treatments with PPF extract on the UV-induced skin responses exhibited similar patterns. In both experiments, the mixture of PEA and PBT significantly inhibited the UV-induced skin and epidermal thickening, while PET inhibited only the UV-induced epidermal thickening. Treatment of PET or the mixture of PEA and PBT significantly inhibited the UV-induced MMP-13 expression, but not type I collagen expression. Topical treatment of the mixture of PEA and PBT with UV irradiation significantly elevated catalase, superoxide dismutase (SOD) and glutathione-peroxidase (GPx) activities in the skin compared to those in the UV irradiated control group, while oral treatment of the mixture of PEA and PBT or PET elevated only catalase and SOD activities, but not GPx. Thirteen phytochemical compounds including 4-O-caffeoylquinic acid, cimicifugic acid E and B, quercetin-3-O-rhamnoside and kaempferol glycoside derivatives were identified in the PPF extract. CONCLUSIONS: These results demonstrate that treatment with PET or the mixture of PEA and PBT, both topically or orally, attenuates UV-induced photoaging via the cooperative interactions of phenolic components having anti-oxidative and collagen-protective activities.

Rational and efficient approach to the preparation of the active fractions of Scutellaria baicalensis (황금(Scutellaria baicalensis) 유효분획물 제조의 합리적이고 효율적인 접근방법)

  • Kim, Doo-Young;Kim, Won Jun;Kim, Jung-Hee;Oh, Sei-Ryang;Ryu, Hyung Won
    • Journal of Applied Biological Chemistry
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    • v.62 no.1
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    • pp.31-38
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    • 2019
  • Scutellaria baicalensis Georgi (Scutellariae Radix) has been widely used as a dietary ingredient and traditional herbal medicine such as diuretic, hyperlipidemia, antibacterial, anti-allergy, anti-inflammatory and anticancer properties. In this study, the isolation of biomarkers or bioactive compounds from complex S. baicalensis extracts represents an essential step for de novo identification and bioactivity assessment. The bioactive fraction consisted of eight compounds which was chromatographed on an analytical high performance liquid chromatography column using two different gradient runs. A simulative replacement of the analytical column with a medium pressure liquid chromatography and open column allowed the determination of gradient profile to allow sufficient separation in the preparative scale. From the optimized method, eight standard compounds have been identified in the fractions. In addition, MS, UV, HRMS detection was provided by ultraperformance liquid chromatographyequadrupole time-of-flight mass spectrometry (UPLC-QTof-MS) of all fractions. Therefore, this scale up procedure was successfully applied to a S. baicalensis extract.

Simultaneous Analysis of Multi-Residual Pesticides Using UPLC (UPLC를 이용한 잔류농약 다성분 동시분석 연구)

  • Kim, Ki-Yu;Eom, Mi-Na;Kang, Jung-Bok;Do, Young-Suk;Moon, Su-Gyeong;Jung, Il-Hyeong;Son, Jong-Sung;Pi, Il-Seon;Park, Eun-Mi;Kim, Young-Su;Lee, Seong-Bong;Cho, Sang-Hun;Moon, Sun-Ae
    • Bulletin of Food Technology
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    • v.21 no.2
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    • pp.49-54
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    • 2008
  • 농산물 중 잔류농약을 보다 쉽고 효율적으로 분석 할 수 있는 다성분 동시분석 방법에 따라서 UPLC-PDA을 이용하여 기기조건에 따른 검출한계 및 정량한계를 식품별 최저 MRL과 비교하여 유효성을 알아보고 회수율을 통하여 방법의 재현성을 조사한 결과 다음과 같다. 41종의 농약에 대한 검량선은 직선성이 우수하였으며 검출한계 $0.005{\sim}0.109mg/kg$으로 나타났으며 정량한계 는 $0.018{\sim}0.363ppm$으로 나타났다. Cycloprothrin 등의 경우처럼 검출기의 감응도가 최저 MRL 기준을 초과하는 농약에 대한 보완으로는 검출감도가보다 우수한 UV 검출기 등의 병행 활용이 필요하다. 회수율에 따른 재현성은 쌀의 경우 첨가 농도 0.4 mg/kg 일때 $62.09{\sim}99.58%$로 편차범위는 $1.03{\sim}13.92%$ 얻을 수 있었다.

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Identification of process generating formaldehyde in a furniture manufacturer (특정 가구 제조 공장의 포름알데히드 발생 공정 노출 평가)

  • Yoo, Kye-Mook;Lee, Mi-Young
    • Analytical Science and Technology
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    • v.27 no.5
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    • pp.243-247
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    • 2014
  • Formaldehyde is defined as carcinogen causing leukaemia, lymphoma or nasopharyngeal carcinoma at high level of exposure. Furniture-manufacturing workers can be exposed to formaldehyde, which implies serious impact on health of the workers. The authors carried out ambient monitoring of formaldehyde in the field, and identified the source of formaldehyde generated during the working process by testing the condition in the laboratory settings. After sampling formaldehyde in the air with 2,4-DNPH (2,4-dinitrophenylhydrazine) coated silica gel, we extracted formaldehyde derivative with acetonitrile and analyzed the extract using HPLC with UV detector at 360 nm. Formaldehyde was separated by ACQUITY UPLC BEH $C_{18}$ column at a flow rate of 0.5 mL/min using 45% acetonitrile as mobile phase. The workers were exposed to higher level of formaldehyde than normal air. Formaldehyde up to 0.31 ppm was detected in the process of veneer attachment, which exceeded 0.3 ppm, the ceiling value of ACGIH standard. The laboratory test of measuring formaldehyde generated from the glue and veneer used in the attachment process resulted in more formaldehyde generation as the temperature increased, and more from the veneer. Heating the veneer to $100-150^{\circ}C$ following the real condition of the manufacturing site generated 1.14-2.70 ppm of formaldehyde from the sample, which was 2-5 times higher level than Korean limit of exposure (0.5 ppm). As the workers handling and processing the veneer which was produced by wet process had high possibility to be exposed to formaldehyde, urgent improvement and management of working environment of furniture manufacturer is demanded.

Analytical Characterization of Aza-Indole Alkaloids in the Biosynthesis of Catharanthus Roseus

  • Lee, Hyang-Yeol
    • Journal of the Korean Applied Science and Technology
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    • v.29 no.2
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    • pp.248-256
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    • 2012
  • Aza-indoles are important pharmacophores that have similar size and biological properties of indole. Here we have synthesized 4- and 7-azaindole tryptamines and showed that they are successfully incorporated in the biosynthesis of monoterepene indole alkaloids (MIAs) to form novel azaindole alkaloids by enzymatic reactions of strictosidine synthase(STR) and strictosidine glucosidase(SDG) monitored by UPLC/MS. By using HPLC equipped with a HPLC photo diode array(PDA) detector, each of the UV spectra of azaindole alkaloids was obtained and characterized. When hydrophilicity of azaindole alkaloids was compared, 4-azaindole alkaloids were more hydrophilic than 7-azaindole alkaloids.

A Study of Cutting Methods by Comparing the Contents of Cinnamic acid and Cinnamaldehyde in Different Parts of Cinnamomi Ramulus (Cinnamic acid, Cinnamaldehyde의 부위별 정량을 통한 계지(桂枝)의 음편(飮片) 규격 연구)

  • Kim, Young-Sik;Lee, Guem-San;Kim, Jung-Hoon;Choi, Go-Ya;Jeong, Seung-Il;Cho, Su-In;Ju, Young-Sung;Kim, Hong-Jun
    • The Korea Journal of Herbology
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    • v.26 no.2
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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.