• Title/Summary/Keyword: ultra performance liquid chromatography (UPLC)

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Method validation of marker compounds from Angelicae Dahuricae Radix as functional food ingredients (건강기능식품 원료로서 구릿대의 지표성분 분석법 검증)

  • Bo-Ram Choi;Dahye Yoon;Hyeon Seon Na;Geum-Soog Kim;Kyung-Sook Han;Sookyeong Lee;Dae Young Lee
    • Journal of Applied Biological Chemistry
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    • v.65 no.4
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    • pp.343-348
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    • 2022
  • This study was performed to establish an analytical method for the standardization of Angelicae Dahuricae Radix as a functional ingredient. We established six compounds including oxypeucedanin hydrate (1), byakangelcol (2), oxypeucedanin (3), imperatorin (4), phellopterin (5) and isoimperatorin (6) as marker compounds of Angelicae Dahuricae Radix. An analytical method using Ultra Performance Liquid Chromatography (UPLC) was established and validated for marker compounds of Angelicae Dahuricae Radix. The specificity was confirmed by the chromatogram from UPLC and the value of coefficient determination was also higher than 0.999, indicating high linearity. The relative standard deviation (RSD) and recovery of marker compounds were less than 5% and in the range of 90- 110%, respectively, which means that this method has high accuracy and precision. Therefore, this analytical method could be used as basic data for the development of functional ingredients for health functional food of Angelicae Dahuricae Radix.

Quantitative Analysis of (+)-Catechin and Diosgenin from Smilax china L. Rhizome (토복령에 함유되는 (+)-Catechin과 Diosgenin의 함량분석)

  • Yin, Jun;Yoon, Ki Hoon;Hwang, Yoon Jeong;Lee, Ji Yeon;Shin, Hwa Sup;Lee, Min Won
    • Korean Journal of Pharmacognosy
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    • v.46 no.3
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    • pp.189-194
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    • 2015
  • Validation and contents determination of (+)-catechin and diosgenin from Smilax china L. rhizome (SC) were confirmed using Ultra-Performance Liquid Chromatography (UPLC). (+)-Catechin was isolated from 60% prethanol SC extract and diosgenin was isolated by acid hydrolysis of saponin fraction from 60% prethanol extract of SC. Finally we have established the validation of the isolated compounds [(+)-catechin, diosgenin] and contents determinations of (+)-catechin and diosgenin by UPLC on 60% prethanol extract of SC [(+)-catechin: 0.06878%, diosgenin: 0.48169%], and on hot water extract of SC [(+)-catechin: 0.06584%, diosgenin: 0.42178%].

Preservatives in Domestic and Imported Children's Clay Products (국내 시판 어린이 점토제품 중 보존제 함유량 조사)

  • Jung, Sun Hye;Heo, Jin Yeong;Oh, Ji Hee;Park, Na-Youn;Kho, Younglim
    • Journal of Environmental Health Sciences
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    • v.48 no.1
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    • pp.36-43
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    • 2022
  • Background: Preservatives are used to prevent product deterioration in modeling clay. Parabens, a representative preservative, have been found to be endocrine disruptors and cause skin irritation and allergic reactions. Isothiazolinone preservatives can be irritating to the skin, respiratory tract, and eyes. Thorough investigation and regulation of clay are necessary because clay is marketed to children, who are more sensitive to the toxic effect of chemicals. Objectives: In this study, the presence of 16 preservatives was analyzed in modeling clay and the results were compared with current standards. Methods: A total of 200 samples were collected from 28 children's clay products sold in South Korea (13 from Korea and 15 imported from overseas). Twelve preservatives, such as parabens, were analyzed using high-performance liquid chromatography (HPLC). Isothiazolinone preservatives (chloromethylisothiazolinone; CMIT, methylisothiazolinone; MIT, octylisothiazolinone; OIT, and benzisothiazolinone; BIT) were analyzed using ultra performance liquid chromatography-tandem mass spectrometery (UPLC-MS/MS). Results: Dehydroacetic acid (DHA) was detected the most in the clays at 51.50% (103 cases) detection; 38 cases (median 190.42 ㎍/g) in Korean products and 65 cases (median 169.62 ㎍/g) in Chinese products. CMIT, which is prohibited in Korea, was detected in 14 (median 16.28 ㎍/g) Chinese products. OIT, which has a chemical structure similar to CMIT was found in 28 (median 68.38 ㎍/g) samples in Korean products. Conclusions: The use of CMIT and MIT in children's products is prohibited in Korea and the European Union (EU). The detection of CMIT in Chinese clay products suggests that management is necessary for imported products. It is necessary to review the safety and regulatory status for OIT because OIT was used as a substitute for CMIT and MIT in Korean products.

Development of an UPLC-DAD Method for Simultaneous Analysis of Eight Marker Compounds of Bulhwangeumjeonggi-san (UPLC-DAD를 이용한 불환금정기산의 다성분 동시분석법 개발)

  • Lee, Kyung-hee;Lamichhane, Ramakanta;Kumar, Sharma Dipak;Raj, Pandeya Prakash;Kim, Se-Gun;Jung, Hyun-Ju
    • Korean Journal of Pharmacognosy
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    • v.47 no.4
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    • pp.366-373
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    • 2016
  • Bulhwangeumjeonggisan (BHGJGS) is a traditional herbal formulation generally used in the treatment of cold and gastritis. BHGJGS consists of eight herbal plants; Atractylodis Rhizoma, Magnoliae Cortex, Citri Pericarpium, Glycyrrhizae Radix, Agastachis Herba, Pinelliae Rhizoma, Zingiberis Rhizoma and Zizyphi Fructus. Complete standardization of this formulation has not been done yet. So, a simple and accurate method was developed and validated using Ultra Performance Liquid Chromatography (UPLC) with Diode Array Detector (DAD) for the standardization of BHGJGS. UPLC conditions were optimized using a c18 RP-Amide column with mobile phase; 0.1% phosphate buffer and acetonitrile, detection wavelength; 210 and 325 nm. The linearities of calibration curves were acceptable ($R^2$>0.9994), and the limit of detection and quantification were within the ranges of 0.011-0.091 and $0.034-0.277{\mu}g/ml$ respectively. The relative standard deviation (RSD) of intra- and inter-day precisions were under 3.61%. The RSD of repeatability was under 0.68 %. The results of recovery test were 94.4-107.9%, and the RSD were under 4.6%. The developed method was used to find the contents of standard constituents in BHGJGS mix extract powder, and two commercial formulation (A and B). The data show that the developed method was specific, sensitive, accurate, and precise for analysis of BHGJGS components.

Establishment of UPLC method for analysis of liquiritigenin and studies on the processing of licorice for enhancement of liquiritigenin content (감초 지표성분 분석법 확립 및 liquiritigenin의 함량 증대를 위한 감초의 수치에 관한 연구)

  • Lee, Jong-Rok;Jo, Mi-Jeong;Park, Sang-Mi;Kim, Sang-Chan;Park, Sook-Jahr
    • Herbal Formula Science
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    • v.18 no.1
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    • pp.145-154
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    • 2010
  • Objective : Licorice has been used for treating digestive disorder and also recommended as a detoxification agent. Liquiritigenin, a component of licorice, has been reported to have various biological activities. In this study, we aimed to establish the analytical method for liquiritigenin content in licorice and the processing method for the enhancement of liquiritigenin content in licorice. Methods : Processing was accomplished by roasting licorice at $250^{\circ}C$ for indicated time periods (5-20 min). Analysis of liquiritigrnin from roasted licorice was conducted using UPLC(Ultra Performance Liquid Chromatography). Results : We established UPLC method for the analysis of liquiritigenin using water : acetonitrile gradient as mobile phase. Furthermore, we standardized the processing condition of licorice to enhance liquiritigenin content using UPLC method. Processing of licorice was accomplished by roasting at $250^{\circ}C$ for indicated time periods (5-20 min) and by pretreating with 50% of acetic acid or 30% ethanol for 24 h. By roasting licorice, the liquiritigenin contents in the licorice were increased. The best roasting time of licorice was 6 min, while roasting for the time above 8 min resulted in diminishing liquiritigenin contents. Moreover, pretreatment with 50% of acetic acid or 30% ethanol picked up liquiritigenin contents in roasted licorice. Conclusion : The adequate processing condition of licorice for the enhancement of liquiritigenin contents was obtained by pretreating licorice with 50% of acetic acid or 30% ethanol for 24 h and then by roasting at $250^{\circ}C$ for 6 min.

Simultaneous Determination of the Flavonoids and Limonoids in Citrus junos Seed Shells Using a UPLC-DAD-ESI/MS

  • Jo, Ara;Shin, Ji hun;Song, Hwa young;Lee, Ye Eun;Jeong, Da Eun;Oh, Sung Hwa;Mun, Myung Jae;Lee, Mina
    • Natural Product Sciences
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    • v.26 no.1
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    • pp.64-70
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    • 2020
  • Citrus junos seeds (CS) have been traditionally used for the treatment of cancer and neuralgia. They are also used to manufacture edible oil and cosmetic perfume. A large amount of CS shells without oil (CSS) are discarded after the oil in CS is used as foods or herbal remedy. To efficiently utilize CSS as a by-products, it needs to be studied through chemical analysis. Therefore, we developed an ultra-performance liquid chromatography (UPLC)-diode array detection (DAD) method for simultaneous determination and quantitative analysis of five components (two flavonoids and threes limonoids) in CSS. A Waters Acquity UPLC HSS T3 column C18 (2.1 × 100 mm, 1.8 ㎛) was used for this separation. It was maintained at 40 ℃. The mobile phase used for the analysis was distilled water and acetonitrile with gradient elution. To identify the quantity of the five components, a mass spectrometer (MS) with an electrospray ionization (ESI) source was used. The regression equation showed great linearity, with correlation coefficient ≥ 0.9912. Limits of detection (LOD) and limits of quantification (LOQ) of the five compounds were 0.09 - 0.13 and 0.26 - 0.38 ㎍/mL, respectively. Recoveries of extraction ranged from 97.45% to 101.91%. Relative standard deviation (RSD) values of intra- and inter-day precision were 0.06 - 1.15% and 0.19 - 0.25%, respectively. This UPLC-DAD method can be validated to simultaneously analyze quantities of marker flavonoids and limonoids in CSS.

Selecting marker substances of main producing area of Codonopsis lanceolata in Korea using UPLC-QTOF-MS analysis (UPLC-QTOF-MS분석를 이용한 국내산 더덕 주산지의 표지물질 선정)

  • An, Young Min;Jang, Hyun-Jae;Kim, Doo-Young;Baek, Nam-In;Oh, Sei-Ryang;Lee, Dae Young;Ryu, Hyung Won
    • Journal of Applied Biological Chemistry
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    • v.64 no.3
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    • pp.245-251
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    • 2021
  • Codonopsis lanceolata (Deoduk) was grown in East Asia, including Korea, China, Japan, and Russia, and the roots of C. lanceolata have been used as functional foods and traditional medicine to treat symptoms of cough, bronchitis, asthma, tuberculosis, and dyspepsia. The phytochemicals of C. lanceolata have been reported such as phenylpropanoids, polyacetylenes, saponins, and flavonoids that are involved in pharmacological effects such as anti-obesity, anti-inflammation, anti-tumor, anti-oxidant, and anti-microbial activities. Selecting marker substances of the main producing area by MS-based metabolomics analysis is important to ensure the beneficial effect of C. lanceolata without side-effects because differences in cultivated areas of plants were related not only to the safety of medicinal plants but also to changes in chemical composition and biological efficacy. In our present study, ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry combined with multivariate statistical analysis was applied to recognize the main producing area of C. lanceolata in South Korea. As a result of Principal Component Analysis and loading plot analysis of three groups, Inje (Kangwon-do), Hoengseong (Kangwon-do), and Muju (Jeonlabuk-do), several secondary metabolites of C. lanceolata including tangshenoside I, lancemaside A, and lancemaside G, were suggested as potential marker substances to distinguish the place of main producing area of C. lanceolata.

Effects of Physicochemical and Environmental Factors on Spatial and Temporal Variations in Phytoplankton Pigment and its Community Composition in Jinhae Bay (진해만에서 물리화학적 환경요인이 식물플랑크톤 색소 및 군집조성의 시공간적 변화에 미치는 영향)

  • Na, Sujin;Lee, Jiyoung;Kim, Jeong Bae;Koo, Jun-Ho;Lee, Garam;Hwang, Hyunjin
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.27 no.2
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    • pp.340-354
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    • 2021
  • The aim of this study was to investigate the spatial and temporal distribution of phytoplankton biomass and community composition in Jinhae Bay on the southern coast of Korea. Phytoplankton pigment analysis was conducted using ultra performance liquid chromatography (UPLC) were conducted from April to December 2019 at seven stations. Temperature, salinity, and dissolved oxygen (DO) and inorganic nutrients (dissolved nitrogen, dissolved phosphorus, and orthosilicic acid) were measured to investigate the environmental factors associated with the structure of phytoplankton community. Phytoplankton biomass (Chl-a) was the highest in July (mean 15.4±4.3 ㎍/L) and the lowest in December (mean 3.5±0.6 ㎍/L). Fucoxanthin was the most abundant carotenoid and showed a similar variation pattern to Chl-a, peridinin, and Chl-b. Phytoplankton community composition analysis showed that diatoms were a predominant group with an average abundance of 70 % whereas chlorophytes, cryptophytes, and dinoflagellates often appeared with lower averages. Further, the dominance of diatoms was closely correlated with water temperature and N:P ratio, which might be influenced by high temperatures in the summer and nutrient loading from the land. Additionally, freshwater and nutrient input by rainfall was estimated to be the most important environmental factor. Hence, the spatial and temporal variations in the composition of phytoplankton pigments and phytoplankton community were correlated with physicochemical and environmental parameters.

Quality inspections of active components in oxidative hair coloring products by UPLC (UPLC를 이용한 염모제품에 함유된 유효성분에 대한 품질검사)

  • Kim, Dong-Gyu;Kim, Yoo-Kyung;Yun, Eun-Sun;Hong, Mi-Sun;Shin, Ji-Young;Jeong, Yoon-Kyung;Kim, Jung-Hun;Chae, Young-Zoo;Park, Seung-Kook
    • Analytical Science and Technology
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    • v.26 no.1
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    • pp.99-105
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    • 2013
  • To identify oxidative hair dyes in hair-coloring products, the thin-layer chromatography (TLC) screening method was used in accordance with Korean Quasi-drug Codex. However, the TLC method is not reliable when there are very small amount of materials to be tested or when $R_f$ values of several components are similar. In this study, Ultra Performance Liquid Chromatography (UPLC) with a rapid sample preparation method was developed for the reliable and sensitive identification of active components contained in oxidative hair-coloring products. Hexane-distilled water was used for the extraction of active components contained in the products prior to UPLC analysis. The limit of detection of active components was 6.7-77.9 ${\mu}g/L$, and the limit of quantitation was 22.3-259.7 ${\mu}g/L$. Except for ${\alpha}$-naphthol, the range of recovery ratio was 96.2-101.5%. From this study, we demonstrated that oxidative active hair-coloring components can easily be analyzed by rapid extraction method followed by UPLC analysis.

Quantitative Analysis of the Marker Constituents in Yongdamsagan-Tang using Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry (LC-ESI-MS/MS를 이용한 용담사간탕의 주요 성분 분석)

  • Seo, Chang-Seob;Ha, Hyekyung
    • Korean Journal of Pharmacognosy
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    • v.48 no.4
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    • pp.320-328
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    • 2017
  • Yongdamsagan-tang has been used to treat the urinary disorders, acute- and chronic-urethritis, and cystitis in Korea. In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS) method was established for simultaneous analysis of the 20 bioactive marker compounds, geniposidic acid, chlorogenic acid, geniposide, liquiritin apioside, acteoside, calceolarioside B, liquiritin, nodakenin, baicalin, liquiritigenin, wogonoside, baicalein, glycyrrhizin, wogonin, glycyrrhizin, wogonin, saikosaponin A, decursin, decursinol angelate, alisol B, alisol B acetate, and pachymic acid in traditional herbal formula, Yongdamsagan-tang. Chromatographic separations of all marker compounds were conducted using a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) at $45^{\circ}C$ using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient elution. The MS analysis was performed using a Waters ACQUITY TQD LC-MS/MS coupled with an electrospray ionization source in the positive and negative modes. The flow rate was 0.3 mL/min and injection volume was $2.0{\mu}L$. The correlation coefficient of 20 marker compounds in the test ranges was 0.9943-1.0000. The limits of detection and quantification values of the all marker components were 0.11-6.66 and 0.34-19.99 ng/mL, respectively. As a result of the analysis using the optimized LC-ESI-MS/MS method, three compounds, geniposidic acid (from Plantaginis Semen), alisol B (from Alismatis Rhizoma), and pachymic acid (from Poria Sclerotium), were not detected in this sample. While the amounts of the 17 compounds except for the geniposidic acid, alisol B, and pachymic acid were $0.04-548.13{\mu}g/g$ in Yongdamsagan-tang sample. Among these compounds, baicalin, bioactive marker compound of Scutellariae Radix, was detected at the highest amount as a $548.13{\mu}g/g$.