• Title/Summary/Keyword: method detection limit

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Rapid Detection for Salmonella spp. by Ultrafast Real-time PCR Assay (Ultrafast Real-time PCR법을 이용한 살모넬라의 신속 검출)

  • Kim, Seok Hwan;Lee, Yu-Si;Joo, In-Sun;Kwak, Hyo Sun;Chung, Gyung Tae;Kim, Soon Han
    • Journal of Food Hygiene and Safety
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    • v.33 no.1
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    • pp.50-57
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    • 2018
  • Salmonella continue to be a major cause of food poisoning worldwide. The rapid detection method of food-borne Salmonella is an important food safety tool. A real-time polymerase chain reaction (PCR) has been used as a rapid method for the detection of pathogens. It has been recently reported that NBS LabChip real-time PCR is a novel, ultrafast, and chip-type-convenient real-time PCR system. We developed the assay method based on NBS LabChip real-time PCR for the rapid detection of Salmonella, which its reaction time was within 20 minutes. Two target genes (invA and stn) were selected to design target specific primers and probes. The new method was validated by checking specificity and sensitivity (limit of detection). This study included forty-two target and twenty-one non-target strains to assess the specificity. This assay was able to identify the 42 Salmonella strains correctly. The limit of detection (LOD) was $10^1copies/{\mu}L$ in Salmonella genomes DNA, while LOD incubated for 4 hr in the inoculated sausage sample ranged from $10^1CFU/g$ to $10^2CFU/g$ as an inoculated cell count. The assay developed in this study could be applied for the investigation of food poisoning pathogens.

Analytical Characteristics of GC/MS and HPLC according to the Concentration Distribution of PAHs (PAHs 농도 분포에 따른 GC/MS와 HPLC의 분석특성에 관한 연구)

  • Hong, Jwa-Ryung;Choi, Kwang-Min
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.25 no.3
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    • pp.312-321
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    • 2015
  • Objectives: The purpose of this study was to determine the best method to analyze PAHs at extremely low concentrations. To this end, 16 PAHswere analyzed simultaneously by GC/MS, HPLC/FLD and HPLC/UVD, and the analytical characteristics of HPLC and GC/MS were compared. Methods: This study was conducted by GC/MS and HPLC/FLD/UVD, and evaluated linearity, precision and detection limit. Standard solutions were prepared for 21 samples in the range of $0.00001{\sim}1.0{\mu}g/mL$ and the samples were divided into four groups. All samples were made in three sets and analysis was replicated seven times. Results: Sixteen PAHs could be simultaneously separated by HPLC and GC/MS, and the adequate equipment was HPLC/FLD. The retention times by HPLC were shorter than GC/MS, and HPLC had better separation for most PAHs than GC/MS. The peaks of naphthalene and naphthalene-D8 partially overlapped for GC/MS. HPLC/FLD had a 20-2000 times lower limit of detection than GC/MS and UVD. However FLD was not adequate for analyzing acenaphthylene because it has too low a fluorescence quantum yield to be detected. The precision of HPLC/FLD/UVD and GC/MS showed less than 20% at $0.001{\mu}g/mL$ PAHs and when the concentration was higher, the coefficient of variation was decreased. HPLC/FLD was better for the overall detection of limits. Conclusions: The results indicate that the HPLC/FLD method has good linear range, precision and a detection of limits from $0.00001{\sim}0.0001{\mu}g/mL$ for all 16 PAHs. This study contributes to providing useful data for analysis technology and can be applied to occupational exposure measurement for PAHs in workplaces.

Methods for the Extraction of DNA from Water Samples for Polymerase Chain Reaction

  • Jung, Jae-Sung;Lee, Young-Jong
    • Journal of Microbiology
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    • v.35 no.4
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    • pp.354-359
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    • 1997
  • Methods for the extraction of DNA from water sample were approximated. Four different procedures of DNA extraction were carried out with pellets obtained from centrifugation of 4 liter water samples. The recovery efficiency and purity of DNA extracted by each method from different sources were compared. DNA yield varied with extraction methods, Method I, which involves enzymatic and freeze-thaw lysis steps and phenol and phenol-chloroform purification of extracted nucleic acid, showed a significantly higher yield and purity than the other methods. The use of glass beads in the DNA extraction methods improved the purity of DNA suitable for PCR. Bovine serum albumin in the PCR reaction mixture was useful in reducing inhibitory effects of contaminants. The efficiency of an extraction method was determined by the detection of the aer of Aeromonas hydrophila with PCR. The lower limit of detection of A. hydrophila from seeded tap water was 2 CFU/ml in PCR when method I was used for DNA preparation.

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Nanowell Array based Sensor and Its Packaging

  • Lee, JuKyung;Akira, Tsuda;Jeong, Myung Yung;Lee, Hea Yeon
    • Journal of the Microelectronics and Packaging Society
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    • v.21 no.3
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    • pp.19-24
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    • 2014
  • This article reviews the recent progress in nanowell array biosensors that use the label-free detection protocol, and are detected in their natural forms. These nanowell array biosensors are fabricated by nanofabrication technologies that should be useful for developing highly sensitive and selective also reproducible biosensors. Moreover, electrochemical method was selected as analysis method that has high sensitivity compared with other analysis. Finally, highly sensitive nanobiosensor was achieved by combining nanofabrication technologies and classical electrochemical method. Many examples are mentioned about the sensing performance of nanowell array biosensors will be evaluated in terms of sensitivity and detection limit compared with other micro-sized electrode without nanowell array.

Simultaneous Determination of Abamectin and Milbemectin Residues in Fruits

  • Lee, Young-Deuk;Kwon, Chan-Hyeok
    • Journal of Applied Biological Chemistry
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    • v.43 no.2
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    • pp.94-100
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    • 2000
  • An analytical method was developed to determine abamectin and milbemectin residues in apple, pear, and citrus using HPLC with ultraviolet absorption detection. Abamectin and milbemectin were extracted with methanol from apple, pear, and citrus samples. The extract was diluted with saline water and dichloromethane partition was followed to recover the compounds from the aqueous phase. Florisil column chromatography and aminopropyl solid-phase extraction were employed as cleanup methods to remove most of co-extractives from the sample extract. Reverse-phase HPLC using an octadecylsilyl column was successfully applied to separate and quantitate abamectin and milbemectin residues in sample extracts at the wavelength of 245 nm. Recoveries of abamectin and milbemectin from fortified samples ranged 80.4~90.3% and 90.9~96.8%, respectively. Relative standard deviations of the analytical method were less than 10% for both acaricides. Detection limit of the analytical method was 0.003 mg/kg sample for all the analytes. The proposed method was reproducible and sensitive enough to evaluate terminal residues of abamectin and milbemectin in apple, pear, and citrus.

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Development of Analytical Method for Polychlorinated Biphenyls and Organochlorine Pesticides in Air (대기 중 PCBs와 유기염소계 살충제의 정량을 위한 분석 방법 개발)

  • 최민규;여현규;김태욱;천만영;선우영
    • Journal of Korean Society for Atmospheric Environment
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    • v.18 no.6
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    • pp.475-485
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    • 2002
  • An analytical method was investigated for the meaiiurement of polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) concentrations in air samples. Procedures required for column chromatographic clean up. silicagel (stage I) and gel permeation chromatography (stage II), were discussed. Identification and quantification of PCBs and OCPs were performed using a combination of gas chromatography/mass spectrometry/ selected ion monitoring. Recovery tests calculated from six samples are 68∼137% for PCBs and 58∼130% for OCPs except for endrin aldehyde. Instrumental detection limits determined for the PCBs and OCPs varied from 0.05 to 0.18 pg/m3 and from 0.71 to 16.82 pg/㎥, respectively. The method has been applied to the analysis of air samples collected at Ansung city, Kyonggi province, Korea. This method may serve as a screening protocol for the simultaneous determination of PCBs and OCPs in air.

Quantification of Three Prohibited Anabolic-Androgenic Steroids in Equine Urine using Gas Chromatography-Tandem Mass Spectrometry

  • Young Beom Kwak;Shaheed Ur Rehman;Hye Hyun, Yoo
    • Mass Spectrometry Letters
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    • v.14 no.3
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    • pp.104-109
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    • 2023
  • Anabolic-androgenic steroids (AAS) are used illegally to enhance muscle development and increase strength and power. In this study, a reliable, and sensitive quantitative method was developed and validated using heptafluorobutyric acid anhydride (HFPA) derivatives for the simultaneous detection of prohibited AAS (testosterone [TS], boldenone [BD], 5α-estrane-3β,17α-diol [EAD]) using gas chromatography-tandem mass spectrometry (GC-MS/MS). For processing the samples, solid phase extraction, methanolic hydrolysis, and liquid-liquid extraction were used. For detection using mass spectrometry, the multiple reaction monitoring (MRM) mode was used with the electron ionization (EI) positive mode. The method was evaluated for selectivity, linearity, lower limit of quantification, intra- and inter-day precision, accuracy, and stability. The results showed that the method was accurate and reproducible for the quantitation of the three steroids. The developed method was finally applied to the analysis of a suspect gelding urine sample received from the Asian Quality Assurance Program (AQAP).

Sample Preparation and Nucleic Acid-based Technologies for the Detection of Foodborne Pathogens (식중독균의 검출을 위한 시료전처리 및 핵산기반의 분석기술)

  • Lim, Min-Cheol;Kim, Young-Rok
    • Food Engineering Progress
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    • v.21 no.3
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    • pp.191-200
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    • 2017
  • There have been great efforts to develop a rapid and sensitive detection method to monitor the presence of pathogenic bacteria in food. While a number of methods have been reported for bacterial detection with a detection limit to a single digit, most of them are suitable only for the bacteria in pure culture or buffered solution. On the other hand, foods are composed of highly complicated matrices containing carbohydrate, fat, protein, fibers, and many other components whose composition varies from one food to the other. Furthermore, many components in food interfere with the downstream detection process, which significantly affect the sensitivity and selectivity of the detection. Therefore, isolating and concentrating the target pathogenic bacteria from food matrices are of importance to enhance the detection power of the system. The present review provides an introduction to the representative sample preparation strategies to isolate target pathogenic bacteria from food sample. We further describe the nucleic acid-based detection methods, such as PCR, real-time PCR, NASBA, RCA, LCR, and LAMP. Nucleic acid-based methods are by far the most sensitive and effective for the detection of a low number of target pathogens whose performance is greatly improved by combining with the sample preparation methods.

Development of Analytical Technology Using the HS-SPME-GC/FID for Monitoring Aromatic Solvents in Urine

  • Lee, Mi-Young;Chung, Yun Kyung;Shin, Kyong-Sok
    • Mass Spectrometry Letters
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    • v.4 no.1
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    • pp.18-20
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    • 2013
  • Headspace solid phase micro-extraction gas chromatography/flame ionization detection (HS-SPME-GC/FID) method was compared with headspace gas chromatography/mass selective detection (HS-GC/MS). Organic solvent-spiked urine as well as urine samples from workspace was analyzed under optimal condition of each method. Detection limit of each compound by HS-SPME-GC/FID was $3.4-9.5{\mu}g/L$, which enabled trace analysis of organic solvents in urine. Linear range of each organic solvent was $10-400{\mu}g/L$, with fair correlation coefficient between 0.992 and 0.999. The detection sensitivity was 4 times better than HS-GC/MS in selected ion monitoring (SIM) mode. Accuracy and precision was confirmed using commercial reference material, with accuracy around 90% and precision less than 4.6% of coefficient of variance. Among 48 urine samples from workplace, toluene was detected from 45 samples in the range of $20-324{\mu}g/L$, but no other solvents were found. As a method for trace analysis, SPME HS GC/FID showed high sensitivity for biological monitoring of organic solvent in urine.

A Method for Preemptive Intrusion Detection and Protection Against DDoS Attacks (DDoS 공격에 대한 선제적 침입 탐지·차단 방안)

  • Kim, Dae Hwan;Lee, Soo Jin
    • Journal of Information Technology Services
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    • v.15 no.2
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    • pp.157-167
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    • 2016
  • Task environment for enterprises and public institutions are moving into cyberspace-based environment and structing the LTE wireless network. The applications "App" operated in the LTE wireless network are mostly being developed with Android-based. But Android-based malwares are surging and they are the potential DDoS attacks. DDoS attack is a major information security threat and a means of cyber attacks. DDoS attacks are difficult to detect in advance and to defense effectively. To this end, a DMZ is set up in front of a network infrastructure and a particular server for defensive information security. Because There is the proliferation of mobile devices and apps, and the activation of android diversify DDoS attack methods. a DMZ is a limit to detect and to protect against DDoS attacks. This paper proposes an information security method to detect and Protect DDoS attacks from the terminal phase using a Preemptive military strategy concept. and then DDoS attack detection and protection app is implemented and proved its effectiveness by reducing web service request and memory usage. DDoS attack detection and protecting will ensure the efficiency of the mobile network resources. This method is necessary for a continuous usage of a wireless network environment for the national security and disaster control.