• Title/Summary/Keyword: Relative quantification

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Relative Quantification of Glycans by Metabolic Isotope Labeling with Isotope Glucose in Aspergillus niger

  • Choi, Soo-Hyun;Cho, Ye-Eun;Kim, Do-Hyun;Kim, Jin-il;Yun, Jihee;Jo, Jae-Yoon;Lim, Jae-Min
    • Mass Spectrometry Letters
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    • v.13 no.4
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    • pp.139-145
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    • 2022
  • Protein glycosylation is a common post-translational modification by non-template-based biosynthesis. In fungal biotechnology, which has great applications in pharmaceuticals and industries, the importance of research on fungal glycoproteins and glycans is accelerating. In particular, the importance of quantitative analysis of fungal glycans is emerging in research on the production of filamentous fungal proteins by genetic modification. Reliable mass spectrometry-based techniques for quantitative glycomics have evolved into chemical, enzymatic, and metabolic stable isotope labeling methods. In this study, we intend to expand quantitative glycomics by metabolic isotope labeling of glycans in Aspergillus niger, a filamentous fungus model, by the MILPIG method. We demonstrate that incubation of filamentous fungi in a culture medium with carbon-13 labeled glucose (1-13C1) efficiently incorporates carbon-13 into N-linked glycans. In addition, for quantitative validation of this method, light and heavy glycans are mixed 1:1 to show the performance of quantitative analysis of various N-linked glycans simultaneously. We have successfully quantified fungal glycans by MILPIG and expect it to be widely applicable to glycan expression levels under various biological conditions in fungi.

Establishment of analytical methods for allergenic compounds in mouthwashes and sanitary napkins by ultra-high-performance liquid chromatography with tandem mass spectrometry

  • Hee-Jung Sim;Hee-Jin Jeong;Yeong-In Lee;Yu-Jin Cho;Seung-Hoon Baek;Jong-Hwan Kim
    • Analytical Science and Technology
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    • v.36 no.2
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    • pp.89-98
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    • 2023
  • Analytical methods for detecting atranol, chloroatranol, evernic acid, (+)-usnic acid, and atranorin in sanitary napkins and mouthwashes were developed using ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). UHPLC-MS/MS conditions were optimized for rapid, sensitive, and simultaneous analysis of the five allergenic compounds. The methods were validated by assessing their specificity, matrix effects, limit of detection (LOD), limit of quantification (LOQ), linearity, accuracy, and precision. Good linearity was achieved with a determination coefficient of ≥0.99. The LOD and LOQ were 2.1-9.8 and 6.4-29.6 ng/g for sanitary napkins and 0.29-0.48 and 0.87-1.45 ng/mL for mouthwashes, respectively. The accuracy and precision were within an acceptable range according to the criteria reported in the European SANTE/11813/2017 guidelines (70-120 % recovery, <20 % relative standard deviation). Therefore, these methods can be used to analyze atranol, chloroatranol, evernic acid, (+)-usnic acid, and atranorin in sanitary napkins and mouthwashes.

Optimization and validation of HPLC/DAD method for the determination of adenosine and cordycepin in cordyceps products

  • Sasikarn Panpraneecharoen;Tisorn Chatrakoon;Sompong Sansenya;Saowapa Chumanee
    • Analytical Science and Technology
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    • v.36 no.4
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    • pp.152-160
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    • 2023
  • Adenosine and cordycepin are bioactive compounds with health benefits. Therefore, both substances are often used to assess the quality of Cordyceps products. Optimization and validation of the HPLC/DAD method for determining two nucleosides were studied. The samples were prepared using an ultrasound-assisted extraction (ultrasonic bath). The result was optimal conditions for aqueous extraction, an extraction time of 35 min, and an extraction temperature of 40 ℃. The Chromatographic separation was achieved using a reverse phase column (InfinityLab Poroshell 120 EC-C18, 4.6 × 250 mm, 2.7 ㎛) at 30 ℃ with a mobile phase gradient elution of water and methanol at a flow rate of 0.7 mL/min. The eluents were monitored via a diode array detector at 260 nm. Two nucleosides were separated by less than 12 min after injection. The developed method was found to be excellent linear (r2 > 0.9999), accurate (% recovery 95.34-98.51), and precise (% relative standard deviation < 2.0). The limit of detection (LOD) and quantification (LOQ) were 0.45 and 1.38 mg/mL for adenosine and 0.47 and 1.43 mg/mL for cordycepin, respectively. This method was satisfactory for simultaneously quantitating two nucleoside contents, which were used to evaluate Cordyceps products.

Development and Validation of a Unique HPLC-ELSD Method for Analysis of 1-Deoxynojirimycin Derived from Silkworms (누에에 함유된 1-Deoxynojirimycin의 분석을 위한 HPLC-ELSD 분석법 밸리데이션)

  • Hyejin Cho;Sullim Lee;Myoung-Sook Shin;Joohwan Lee;Sanghyun Lee
    • Korean Journal of Pharmacognosy
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    • v.54 no.1
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    • pp.38-43
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    • 2023
  • A simple and accurate assay was developed for the quantitative analysis of 1-deoxynojirimycin (1-DNJ) derived from the silkworm (Bombyx mori). Normal-phase high-performance liquid chromatography coupled with an evaporative light scattering detector (HPLC-ELSD) and a hydrophilic interaction liquid chromatography column was used. Various parameters were applied to optimize the analysis method. The limits of detection and quantification of 1-DNJ were 2.97 × 10-3 and 9.00 × 10-3 mg/mL, respectively. The calibration curve showed good linearity results. The concentration range and the r2 value were 0.0625-1.0 mg/mL and 0.9997, respectively. The accuracy test demonstrated a significantly high recovery rate (89.95-103.22%). The relative standard deviation was ≤ 1.00%. Thus, a method for the accurate identification and quantitative analysis of 1-DNJ in silkworms was developed. Moreover, in this procedure, the process of derivatization of 1-DNJ, which was required in previous experiments, could be eliminated. This technique may be actively utilized for the development of pharmaceuticals and health functional foods using 1-DNJ.

Quantitative analysis of hyperoside and isoquercitrin in methanolic extract of Stewartia koreana leaves using HPLC-DAD

  • Ju-Yeong Kang;Yu Hwa Kim;Youngdae Yoon;Bong-Gyu Kim
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.436-446
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    • 2023
  • Since Stewartia koreana leaves are registered with the Food and Drug Administration as edible herbal materials, they are used in the development of functional foods, cosmetics, and medicines. In this study, we established an analysis method that can simultaneously analyze two indicators, hyperoside (quercetin 3-O-galactoside) and isoquercitrin (quercetin 3-O-glucoside) contained in the leaves of S. koreana using HPLC-DAD. In accordance with the Ministry of Food and Drug Safety's health functional food guidelines, the analysis method was verified for specificity, accuracy, precision, limit of quantification, and linearity. The analysis method established in this study showed more than 0.9989 of the correlation coefficient values (r2) for the calibration. The total recovery rates of isoquercitrin and hyperoside were 100.55 and 98.87% with 0.14-0.78 and 0.47-0.67% of the relative standard deviation, respectively. Therefore, it was suggested that the new analytical method would be applied to standardize raw materials and high value-added products originated from the leaves of the S. koreana in the future.

1H Solid-state NMR Methodology Study for the Quantification of Water Content of Amorphous Silica Nanoparticles Depending on Relative Humidity (상대습도에 따른 비정질 규산염 나노입자의 함수량 정량 분석을 위한 1H 고상 핵자기 공명 분광분석 방법론 연구)

  • Oh, Sol Bi;Kim, Hyun Na
    • Korean Journal of Mineralogy and Petrology
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    • v.34 no.1
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    • pp.31-40
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    • 2021
  • The hydrogen in nominally anhydrous mineral is known to be associated with lattice defects, but it also can exist in the form of water and hydroxyl groups on the large surface of the nanoscale particles. In this study, we investigate the effectiveness of 1H solid-state nuclear magnetic resonance (NMR) spectroscopy as a robust experimental method to quantify the hydrogen atomic environments of amorphous silica nanoparticles with varying relative humidity. Amorphous silica nanoparticles were packed into NMR rotors in a temperature-humidity controlled glove box, then stored in different atmospheric conditions with 25% and 70% relative humidity for 2~10 days until 1H NMR experiments, and a slight difference was observed in 1H NMR spectra. These results indicate that amount of hydrous species in the sample packed in the NMR rotor is rarely changed by the external atmosphere. The amount of hydrogen atom, especially the amount of physisorbed water may vary in the range of ~10% due to the temporal and spatial inhomogeneity of relative humidity in the glove box. The quantitative analysis of 1H NMR spectra shows that the amount of hydrogen atom in amorphous silica nanoparticles linearly increases as the relative humidity increases. These results imply that the sample sealing capability of the NMR rotor is sufficient to preserve the hydrous environments of samples, and is suitable for the quantitative measurement of water content of ultrafine nominally anhydrous minerals depending on the atmospheric relative humidity. We expect that 1H solid-state NMR method is suitable to investigate systematically the effect of surface area and crystallinity on the water content of diverse nano-sized nominally anhydrous minerals with varying relative humidity.

Quantification of a Global Construction Core Competencies for Korean Construction/Engineering Firms (국내 건설업체의 해외 진출역량 계량화 연구)

  • Kim, Sang-Bum;Kim, Yong-Bi
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.33 no.6
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    • pp.2541-2549
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    • 2013
  • The Construction industry has been dealing with much trouble due to global economic recession and domestic political trends emphasizing on welfare than development. Consequently, domestic construction market has been dramatically shrunk during the last a few years, and international market has become the only potential solution for the industry. However, there has been lack of efforts in developing a quantified measure of global competencies for Korean engineering and construction organizations. This study attempted to develop quantified indices for Korean engineering and construction contractors with which the level of global construction competencies can be objectively monitored. In doing so, a survey questionnaire was developed to identify relative importances of core competency elements which were derived from extensive literature reviews and experts interviews. AHP (Analytic Hierarchy Process) was employed as a main analysis method in developing quantification measures. The analysis results reveal little differences in competency requirements between engineering and construction firms and it implies that the global market becomes more integrated and requires a total solution for a construction project. The developed core competency measures can be used to quantify the level of preparedness of Korean engineering and construction firms at the time of evaluation and also be used as a basis for performance benchmarking indicators if they are compared with business showings.

Analysis of Pencycuron in Agricultural Products by Simultaneous Determination (동시 다성분 분석법에 의한 농산물 중 펜시쿠론 분석)

  • Park, Young-Hye;Han, Chang-Ho;Kim, Ae-Kyung;Shin, Jae-Min;Lee, Jae-Kyoo;Park, Sun-Hee;Yun, Eun-Sun;Kim, Mu-Sang;Chae, Young-Zoo
    • The Korean Journal of Pesticide Science
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    • v.17 no.3
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    • pp.162-167
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    • 2013
  • An analytical method for the simultaneous determination of pencycuron in agricultural products was established by using GC-NPD. The method was validated through the guidelines of linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy and precision. The calibration curve of pencycuron was linear over the concentration range of 0.02-2 mg/kg with correlation coefficient of above 0.9999. The limits of detection and quantification were 0.005 and 0.02 mg/kg. Mean recoveries of pencycuron for each sample were 79.6-107.5% at the level of 0.02 mg/kg and 86.4-112.1% at the level of 0.2 mg/kg and 84.0- 104.9% at the level of 1 mg/kg. Relative standard deviation (RSD) in recoveries were all less than 5%. The intra- and inter-day precision (RSD) were 1.6-3.0% and 5.3-11.5%, respectively. The result of validation indicated that this method was accurate and sensitive assay.

HPLC Method for Simultaneous Quantitative Detection of Quercetin and Curcuminoids in Traditional Chinese Medicines

  • Ang, Lee Fung;Yam, Mun Fei;Fung, Yvonne Tan Tze;Kiang, Peh Kok;Darwin, Yusrida
    • Journal of Pharmacopuncture
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    • v.17 no.4
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    • pp.36-49
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    • 2014
  • Objectives: Quercetin and curcuminoids are important bioactive compounds found in many herbs. Previously reported high performance liquid chromatography ultraviolet (HPLC-UV) methods for the detection of quercetin and curcuminoids have several disadvantages, including unsatisfactory separation times and lack of validation according the standard guidelines of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. Methods: A rapid, specific, reversed phase, HPLC-UV method with an isocratic elution of acetonitrile and 2% v/v acetic acid (40% : 60% v/v) (pH 2.6) at a flow rate of 1.3 mL/minutes, a column temperature of $35^{\circ}C$, and ultraviolet (UV) detection at 370 nm was developed. The method was validated and applied to the quantification of different types of market available Chinese medicine extracts, pills and tablets. Results: The method allowed simultaneous determination of quercetin, bisdemethoxycurcumin, demethoxycurcumin and curcumin in the concentration ranges of $0.00488-200{\mu}g/mL$, $0.625-320{\mu}g/mL$, $0.07813-320{\mu}g/mL$ and $0.03906-320{\mu}g/mL$, respectively. The limits of detection and quantification, respectively, were 0.00488 and $0.03906{\mu}g/mL$ for quercetin, 0.62500 and $2.50000{\mu}g/mL$ for bisdemethoxycurcumin, 0.07813 and $0.31250{\mu}g/mL$ for demethoxycurcumin, and 0.03906 and $0.07813{\mu}g/mL$ for curcumin. The percent relative intra day standard deviation (% RSD) values were $0.432-0.806{\mu}g/mL$, $0.576-0.723{\mu}g/mL$, $0.635-0.752{\mu}g/mL$ and $0.655-0.732{\mu}g/mL$ for quercetin, bisdemethoxycurcumin, demethoxycurcumin and curcumin, respectively, and those for intra day precision were $0.323-0.968{\mu}g/mL$, $0.805-0.854{\mu}g/mL$, $0.078-0.844{\mu}g/mL$ and $0.275-0.829{\mu}g/mL$, respectively. The intra day accuracies were 99.589%-100.821%, 98.588%-101.084%, 9.289%-100.88%, and 98.292%-101.022% for quercetin, bisdemethoxycurcumin, demethoxycurcumin and curcumin, respectively, and the inter day accuracy were 99.665%-103.06%, 97.669%-103.513%, 99.569%-103.617%, and 97.929%-103.606%, respectively. Conclusion: The method was found to be simple, accurate and precise and is recommended for routine quality control analysis of commercial Chinese medicine products containing the flour flavonoids as their principle components in the extracts.

Development of Real-time and Simultaneous Quantification of Volatile Organic Compounds in Ambient with SIFT-MS (Selected Ion Flow Tube-Mass Spectrometry) (선택적다중이온질량분석기를 이용한 대기 중 휘발성유기화합물 실시간 동시분석법 개발 및 적용)

  • Son, Hyun Dong;An, Joon Geon;Ha, Sung Yong;Kim, Gi Beum;Yim, Un Hyuk
    • Journal of Korean Society for Atmospheric Environment
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    • v.34 no.3
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    • pp.393-405
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    • 2018
  • Volatile organic compounds (VOCs) are representative air pollutants due to their detrimental effects on human health and their role in formation of secondary organic aerosols. Assessments and monitoring programs of VOCs using periodic grab sampling like Tedlar bags, canisters, and sorbent traps provide limited information, often with delay times of days or weeks. Selected ion flow tube mass spectrometry (SIFT-MS) is an emerging analytical technique for the real-time quantification of VOCs in air. It relies on chemical ionization of the VOCs molecules in air introduced into helium carrier gas using $H_3O^+$, $NO^+$, and $O_2{^+}$ precursor ions. Real-time monitoring method of 60 VOCs in the ambient air was developed using TO-15 standard gas mixture. Calibration curves, method detection limit, and quantitation reproducibility of the target compounds were tested. Dynamic dilution system was used to dilute standard gas from 0.174 ppbv to 100 ppbv, where calibration curves showed good linearity with $r^2$> 0.95 in all target analytes. Limit of detection (LOD) all compounds were sub ppbv, and some halogenated compounds showed pptv levels. Seven consecutive analyses of target compounds showed good repeatability with relative standard deviation of less than 10%. One day monitoring of VOCs in ambient air was conducted in Geoje. Average concentration of target VOCs in Geoje were relatively lower than other regions, among which formaldehyde showed the highest concentration ($15.4{\pm}5.78ppbv$). SIFT-MS provided good temporal resolution data (1 data per 3.2 minute), which can be used for identifying ephemeral short-term event. It is expected that SIFT-MS will be a versatile monitoring platform for VOCs in ambient air.