• 제목/요약/키워드: limit of quantification

검색결과 455건 처리시간 0.027초

Validation of a Real-Time RT-PCR Method to Quantify Newcastle Disease Virus (NDV) Titer and Comparison with Other Quantifiable Methods

  • Jang, Juno;Hong, Sung-Hwan;Kim, Ik-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제21권1호
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    • pp.100-108
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    • 2011
  • A method for the rapid detection and quantification of Newcastle disease virus (NDV) produced in an animal cell culture-based production system was developed to enhance the speed of the NDV vaccine manufacturing process. A SYBR Green I-based real-time RT-PCR was designed with a conventional, inexpensive RT-PCR kit targeting the F gene of the NDV LaSota strain. The method developed in this study was validated for specificity, accuracy, precision, linearity, limit of detection (LOD), limit of quantification (LOQ), and robustness. The validation results satisfied the predetermined acceptance criteria. The validated method was used to quantify virus samples produced in an animal cell culture-based production system. The method was able to quantify the NDV samples from mid- or late-production phases, but not effective on samples from the early-production phase. For comparison with other quantifiable methods, immunoblotting, plaque assay, and tissue culture infectious dose 50 ($TCID_{50}$) assay were also performed with the NDV samples. The results demonstrated that the real-time RT-PCR method is suitable for the rapid quantification of virus particles produced in an animal cell-culture-based production system irrespective of viral infectivity.

환경시료 측정에서 분석자의 숙련도가 분석결과에 미치는 영향 (The Effect of Proficiency in Environmental Sample Measurement on Analysis Results)

  • 윤석표;강성민;손연미;전강원
    • 유기물자원화
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    • 제29권2호
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    • pp.25-30
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    • 2021
  • 본 연구는 인산염 인 농도 측정과 관련하여 숙련도에 의해서 정도관리의 지표가 어떻게 개선되는지를 조사하였다. 아울러 분석기기의 종류에 따른 측정값의 차이가 있는지 함께 비교하였다. 분석자의 숙련도가 분석결과에 미치는 영향을 알아보기 위해 3인의 실험자가 2종의 분석기기를 사용하여 수질오염공정시험기준에 따라 인산염 인을 7회에 걸쳐서 방법검출한계와 정량한계를 측정하였으며, 실험결과 5회 이상 반복실험하면 정도관리지표를 만족하였다. 인산염 인의 정량한계는 0.02 mg/L로 계산되었으며, 분석기기가 다른 경우 정량한계 부근의 시료에 대한 흡광도와 농도가 통계적으로 유의미하게 차이가 있음을 확인하였다.

Sampling Methods for Quantification of Solid-state Phases in Powder Samples with Solid-state NMR Spectroscopy

  • Han, Oc-Hee;Kim, Sun-Ha;Ko, Tae-Jung
    • Bulletin of the Korean Chemical Society
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    • 제30권5호
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    • pp.1077-1079
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    • 2009
  • To minimize the variance in the quantification of solid-state phases in powder samples, gently placing polycrystalline samples one next to another directly in a sample holder is better than trying to mix them homogeneously prior to transferring to a sample holder. However, the solid-state cross polarization-magic angle spinning (CP-MAS) nuclear magnetic resonance (NMR) spectroscopy results demonstrated that it is essential in this sampling method to place all the samples in the location of consistent signal sensitivity. The same sampling method may be employed in other spectroscopic quantification techniques of solid-state phases if the method to limit the sample to the location with uniform signal sensitivity in the sample holder is adapted to each technique.

Validation of an HPLC/UV-based method for Salicornia herbacea-derived isorhamnetin-3-O-glucoside and quercetin-3-O-glucoside quantification

  • Park, Jun Yeon;Paje, Leo Adrianne;Kang, Ki Sung;Lee, Sanghyun
    • Journal of Applied Biological Chemistry
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    • 제64권3호
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    • pp.285-290
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    • 2021
  • Salicornia herbacea is a type of salt marsh plant that has been used in traditional medicine to treat several diseases. Isorhamnetin-3-O-glucoside (I3G) and quercetin-3-O-glucoside (Q3G) are major flavonoids in S. herbacea that are known to exert various pharmacological activities. Therefore, our study sought to validate and optimize an HPLC/UV-based analytical method for I3G and Q3G yield quantification, as well as to determine its limit of detection, limit of quantification, linearity, precision, and accuracy. Upon testing a concentration range of 31.5-1.9 ㎍/mL the results exhibited good linearity (r2 ≥0.9996 and r2 ≥0.9999 for I3G and Q3G, respectively), and the procedure was deemed precise (relative standard deviation of ≤3.19 and ≤3.85%, respectively), and accurate (102.6-105.0 and 92.9-95.2%, respectively). The results showed that our proposed method could be used for rapid I3G and Q3G evaluation in S. herbacea.

ANALYSIS OF UNCERTAINTY QUANTIFICATION METHOD BY COMPARING MONTE-CARLO METHOD AND WILKS' FORMULA

  • Lee, Seung Wook;Chung, Bub Dong;Bang, Young-Seok;Bae, Sung Won
    • Nuclear Engineering and Technology
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    • 제46권4호
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    • pp.481-488
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    • 2014
  • An analysis of the uncertainty quantification related to LBLOCA using the Monte-Carlo calculation has been performed and compared with the tolerance level determined by the Wilks' formula. The uncertainty range and distribution of each input parameter associated with the LOCA phenomena were determined based on previous PIRT results and documentation during the BEMUSE project. Calulations were conducted on 3,500 cases within a 2-week CPU time on a 14-PC cluster system. The Monte-Carlo exercise shows that the 95% upper limit PCT value can be obtained well, with a 95% confidence level using the Wilks' formula, although we have to endure a 5% risk of PCT under-prediction. The results also show that the statistical fluctuation of the limit value using Wilks' first-order is as large as the uncertainty value itself. It is therefore desirable to increase the order of the Wilks' formula to be higher than the second-order to estimate the reliable safety margin of the design features. It is also shown that, with its ever increasing computational capability, the Monte-Carlo method is accessible for a nuclear power plant safety analysis within a realistic time frame.

기체크로마토그래피에 의한 한국산 배초향의 정유 분석과 Cholinesterase 억제활성 (Gas Chromatographic Analysis and Cholinesterase Activity of the Essential Oil from Korean Agastache rugosa)

  • 최재수;송병민;박희준
    • 생약학회지
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    • 제47권2호
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    • pp.192-196
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    • 2016
  • The herb of Agastache rugosa (Lamiaceae) called Korean mint as a spice or Agastache Herba as a crude drug is known to contain highly fragrant volatile substances. This research aimed to establish the quantitative gas chromatography (GC) method on the essential oil of A. rugosa using the three standard compounds, estragole, methyleugenol, pulegone, and to find whether the essential oil has anti-Alzheimer's activity. The GC quantification method was established by determining the linearity of calibration curve ($R^2$), linear range, and both limit-of-detection (LOD) and limit-of-quantification (LOQ). The $IC_{50}$ of the essential oil on the activities of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) were determined to be $69.06{\pm}0.26$ and $76.71{\pm}0.58{\mu}g/ml$, respectively.

Different Real Time PCR Approaches for the Fine Quantification of SNP's Alleles in DNA Pools: Assays Development, Characterization and Pre-validation

  • Mattarucchi, Elia;Marsoni, Milena;Binelli, Giorgio;Passi, Alberto;Lo Curto, Francesco;Pasquali, Francesco;Porta, Giovanni
    • BMB Reports
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    • 제38권5호
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    • pp.555-562
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    • 2005
  • Single nucleotide polymorphisms (SNPs) are becoming the most common type of markers used in genetic analysis. In the present report a SNP has been chosen to test the applicability of Real Time PCR to discriminate and quantify SNPs alleles on DNA pools. Amplification Refractory Mutation System (ARMS) and Mismatch Amplification Mutation Assay (MAMA) has been applied. Each assay has been pre-validated testing specificity and performances (linearity, PCR efficiency, interference limit, limit of detection, limit of quantification, precision and accuracy). Both the approaches achieve a precise and accurate estimation of the allele frequencies on pooled DNA samples in the range from 5% to 95% and don't require standard curves or calibrators. The lowest measurement that could be significantly distinguished from the background noise has been determined around the 1% for both the approaches, allowing to extend the range of quantifications from 1% to 99%. Furthermore applicability of Real Time PCR assays for general diagnostic purposes is discussed.

Development and validation of an analytical method for the quantification of 2,6-diisopropylnaphthalene in agricultural products using GC-MS/MS

  • Lee, Han Sol;Park, Ji-Su;Lee, Su Jung;Shin, Hye-Sun;Chung, Yun mi;Choi, Ha na;Yun, Sang Soon;Jung, Yong-hyun;Oh, Jae-Ho
    • 분석과학
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    • 제34권1호
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    • pp.1-8
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    • 2021
  • An analytical method was developed and optimized for the quantification of a plant growth regulator, 2,6-diisopropylnaphthalene (2,6-DIPN), in agricultural products using gas chromatography-tandem mass spectrometry. The samples were extracted, partitioned, and were purified using a Florisil® cartridge. To validate the analytical method, its specificity, linearity, limit of detection (LOD) and limit of quantification (LOQ) of the instrument, LOQ of the analytical method (MLOQ), accuracy, and repeatability were considered. The method displayed excellent results during validation, and is suitable for the determination and quantification of the low residual levels of the analyte in the agricultural samples. All of the results with the optimized method were satisfactory and within the criteria ranges requested in the Codex Alimentarius Commission guidelines and the Ministry of Food and Drug Safety guidelines for pesticide residue analysis. The developed method is simple and accurate and can be used as a basis for safety management of 2,6-DIPN.

Quantification of Allantoin in Yams (Dioscorea sp.) Using a 1H NMR Spectroscopic Method

  • Thao Quyen Cao;Dongyup Hahn
    • Journal of Microbiology and Biotechnology
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    • 제33권5호
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    • pp.662-667
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    • 2023
  • Allantoin is an abundant component of yams and has been known as a skin protectant due to its pharmacological activities. In previous methods for allantoin determination using high-performance liquid chromatography (HPLC), the separation was unsatisfactory. We herein developed a 1H quantitative nuclear magnetic resonance (qNMR) method for quantification of allantoin in the flesh and peel of yams. The method was carried out based on the relative ratio of signals integration of allantoin to a certain amount of the internal standard dimethyl sulfone (DMSO2) and validated in terms of specificity, linearity (range 62.5-2000 ㎍/ml), sensitivity (limit of detection (LOD) and quantification (LOQ) 4.63 and 14.03 ㎍/ml, respectively), precision (RSD% 0.02-0.26), and recovery (86.35-92.11%). The method was then applied for the evaluation of allantoin in flesh and peel extracts of four different yams cultivated in Korea.

A Simple and Sensitive High Performance Liquid Chromatography-Electrospray Ionization/Mass Spectrometry Method for the Quantification of Ethyl Pyruvate in Rat Plasma

  • Kim, Hyun-Ji;Kim, Seung-Woo;Lee, Ja-Kyeong;Yoon, Sung-Hwa
    • Bulletin of the Korean Chemical Society
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    • 제32권4호
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    • pp.1221-1227
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    • 2011
  • Ethyl pyruvate (EP) is known as a scavenger of reactive oxygen species (ROS) in the body through its role in the donation of diketone groups to metals to form an EP-metal complex. In order to develop a method for the quantification of EP in biological media, a sensitive and specific, high-performance liquid chromatographyelectrospray ionization-mass spectrometry (HPLC-ESI/MS) method is used to determine the EP-alkali metal ion binding species. The analyte was separated on a ZORBOX SB-C8 ($3.5{\mu}m$, $30mm{\times}2.1mm$ I.D.) column and analyzed in selected ion monitoring (SIM) mode with a positive ESI interface using the m/z 255 $[2M + Na]^+$ ion. The method was validated over the concentration range of $0.5-60.0\;{\mu}g$/mL under 1/9 (v/v) of acetonitrile/methanol solvent system with flow rate 0.05 mL/min. The limit of quantification (LOQ) was $0.5{\mu}g$/mL.