• 제목/요약/키워드: method detection limit

검색결과 1,352건 처리시간 0.035초

Development of Real-Time PCR for the Detection of Clostridium perfringens in Meats and Vegetables

  • Chon, Jung-Whan;Park, Jong-Seok;Hyeon, Ji-Yeon;Park, Chan-Kyu;Song, Kwang-Young;Hong, Kwang-Won;Hwang, In-Gyun;Kwak, Hyo-Sun;Seo, Kun-Ho
    • Journal of Microbiology and Biotechnology
    • /
    • 제22권4호
    • /
    • pp.530-534
    • /
    • 2012
  • A real-time PCR assay was developed and validated inhouse specifically for the detection of Clostridium perfringens (Cl. perfringens) in meats and vegetables by comparing with the culture method. The detection limit of the real-time PCR assay in phosphate-buffered saline was $10^2$ CFU/ml. When the two methods were compared in food samples inoculated with Cl. perfringens, the culture method detected 52 positives, whereas real-time PCR detected 51 positives out of 160 samples. The difference was without statistical significance (p>0.05). Real-time PCR assay is an option for quality assurance laboratories to perform standard diagnostic tests, considering its detection ability and time-saving efficiency.

Development of a Magnetic Bead-Based Method for Specific Detection of Enterococcus faecalis Using C-Terminal Domain of ECP3 Phage Endolysin

  • Yoon-Jung Choi;Shukho Kim;Jungmin Kim
    • Journal of Microbiology and Biotechnology
    • /
    • 제33권7호
    • /
    • pp.964-972
    • /
    • 2023
  • Bacteriophage endolysins are peptidoglycan hydrolases composed of cell binding domain (CBD) and an enzymatically active domain. A phage endolysin CBD can be used for detecting bacteria owing to its high specificity and sensitivity toward the bacterial cell wall. We aimed to develop a method for detection of Enterococcus faecalis using an endolysin CBD. The gene encoding the CBD of ECP3 phage endolysin was cloned into the Escherichia coli expression vector pET21a. A recombinant protein with a C-terminal 6-His-tag (CBD) was expressed and purified using a His-trap column. CBD was adsorbed onto epoxy magnetic beads (eMBs). The bacterial species specificity and sensitivity of bacterial binding to CBD-eMB complexes were determined using the bacterial colony counting from the magnetic separations after the binding reaction between bacteria and CBD-eMB complexes. E. faecalis could bind to CBD-eMB complexes, but other bacteria (such as Enterococcus faecium, Staphylococcus aureus, Escherichia coli, Acinetobacter baumannii, Streptococcus mutans, and Porphyromonas gingivalis) could not. E. faecalis cells were fixed onto CBD-eMB complexes within 1 h, and >78% of viable E. faecalis cells were recovered. The E. faecalis recovery ratio was not affected by the other bacterial species. The detection limit of the CBD-eMB complex for E. faecalis was >17 CFU/ml. We developed a simple method for the specific detection of E. faecalis using bacteriophage endolysin CBD and MBs. This is the first study to determine that the C-terminal region of ECP3 phage endolysin is a highly specific binding site for E. faecalis among other bacterial species.

Development of Diffusion - Precipitation Method to Determine AVS Concentrations in Freshwater Sediments

  • Song, Ki-Hoon
    • 환경생물
    • /
    • 제23권4호
    • /
    • pp.374-378
    • /
    • 2005
  • A diffusion - precipitation method was developed to determine acid volatile sulfide (AVS) concentrations in freshwater sediments. This method uses silver nitrate as a sulfide trap solution and the concentration of trapped sulfide is determined gravimetrically. The proposed diffusion - precipitation method is more rapid and less expensive than previously developed purge- and - trap methods. Spiked sodium sulfide recoveries using this method $(97\~120\%)$ were similar with a previously developed diffusion - absorption method $(93.8\~115\%)$ and about $20\%$ greater than a previously developed purge-and-trap method $(74.6\~105\%)$. Detection limit of this method $(0.1\;{\mu}mole\;S\;g^{-l})$ was comparable with that of diffusion-absorption method $(0.06\;{\mu}mole\;S\;g^{-l})$ and purge-and-trap method $(0.05\~0.5\;{\mu}mole\;S\;g^{-l})$.

감태추출물의 기능성원료 표준화를 위한 지표성분 dieckol의 분석법 검증 (Validation of an analytical method of dieckol for standardization of Ecklonia cava extract as a functional ingredient)

  • 허연;김은서;이지수;김건희;이현규
    • 한국식품과학회지
    • /
    • 제51권5호
    • /
    • pp.420-424
    • /
    • 2019
  • 본 연구는 해양수산물 유래 기능성 소재인 감태의 지표성분 분석법 검증 및 표준화를 위하여 HPLC를 이용하여 dieckol의 분석법을 확립하고 분석법에 대한 유효성 검증(validation)을 실시하고자 하였다. Dieckol 분석을 위하여 0.1% trifluoroacetic acid가 첨가된 water와 acetonitrile을 이동상으로 Capcell pak $C_{18}$ ($250{\times}4.6mm$, $5.0{\mu}m$) 컬럼을 사용하여 HPLC 방법을 통해 분석한 결과, 표준용액과 감태추출물의 dieckol RT이 일치하고 3.37의 높은 분리도(Rs)를 나타내었으며, UV spectrum이 일치하는 특이성을 확인하였다. 또한 검량선의 상관계수($r^2$)는 0.9994로 높은 직선성을 나타냈으며, 검출한계(LOD)와 정량한계(LOQ)는 0.38과 $1.16{\mu}g/mL$로 각각 확인되었다. 분석법의 재현성을 검증하기 위한 intra-day 및 inter-day 분석에서 RSD는 각각 1.7%와 1.00-1.25%로 나타났으며, 회수율은 99.42-101.32%이였으며 RSD는 2% 이하로 산출되어 정밀성이 있음을 확인하였다. 이상의 결과로부터 HPLC를 이용한 dieckol의 분석법은 감태추출물의 기능성원료 표준화를 위한 지표성분 분석법으로 적합함을 확인하였다. 본 시험법에 따라 분석한 감태추출물 내의 dieckol의 평균 함량은 약 0.11 g/g (10.54%)로 상대표준편차는 0.5% 이하였다. 따라서 이상의 분석결과를 통해 확립된 dieckol 분석법이 향후 감태추출물의 건강기능성식품 기능성 원료 개발과 표준화를 위한 기초자료로 크게 활용될 것으로 기대된다.

총질소 분석방법의 비교 (Comparison of the Total Nitrogen Determination Methods)

  • 정광욱;윤춘경;함종화
    • 한국농공학회:학술대회논문집
    • /
    • 한국농공학회 2001년도 학술발표회 발표논문집
    • /
    • pp.421-425
    • /
    • 2001
  • Among the many parameters describing water quality, total nitrogen content(TN) is regarded as important one in relation to water quality management of the waterbodies for its role in eutrophication process. This study evaluated for accuracy, convenience and promptness of nitrogen analysis methods : (1)UV spectrophotometric method, (2)summation method, (3)cadmium reduction method. And, the detection limit of each method was calculated using EPA method. The UV spectrophotometric method after oxidation appears to be more convenient and accurate in determination of the TN contents than the summation method and cadmium reduction method. The summation method is necessary to separate determinations of the organic nitrogen, ammonia, nitrite, and nitrate. And, because summation method has many other methods for each contents, it can be detected for wide range of concentration.

  • PDF

초음파 Spectroscopy에 의한 두께측정을 위한 다중반사파의 시뮬레이션 (Computer Simulation of Multiple Reflection Waves for Thickness Measurement by Ultrasonic Spectroscopy)

  • 박익근;한응교;최만용
    • 비파괴검사학회지
    • /
    • 제12권1호
    • /
    • pp.9-15
    • /
    • 1992
  • Ultrasonic spectroscopy is likely to become a very powerful NDE method for detection of microfects and thickness measurement of thin film below the limit of ultrasonic distance resolution in the opaque materials, provides a useful information that cannot be obtained by a conventional ultrasonic measuring system. In this paper, we considered a thin film below the limit of ultrasonic distance resolution sandwitched between two substances as acoustical analysis model, demonstrated the usefulness of ultrasonic spectroscopic analysis technique using information of ultrasonic frequency for measurements of thin film thickness, regardless of interference phenomenon and phase reversion of ultrasonic waveform. By using frequency intervals(${\triangle}f$) of periodic minima from the ratio of reference power spectrum of reflective waveform obtained a sample to power spectrum of multiple reflective waves obtained interference phenomenon caused by ultrasonic waves reflected at the upper and lower surfaces of a thin layer, can measured even dimensions of interest are smaller than the ultrasonic wave length with simplicity and accuracy.

  • PDF

식육중 Chlorfluazuron의 잔류분석법 및 잔류량에 관한 연구 (Studies on the Analytical Methods and Quantity of Residual Chlorfluazuron in Meat)

  • 박건상;홍무기;백선영;정병곤;박종세
    • 한국식품위생안전성학회지
    • /
    • 제12권4호
    • /
    • pp.288-293
    • /
    • 1997
  • Analytical method using capillary GC/ECD was developed to determine trace residues of chlofluazuron, 1-[3, 5-dichloro-5-trifluoromethyl-2-pyridyloxy)phenyl]-3-(2, 6-difiuorobenzoyl), in meat, and applied to analyze the residues in domestic and imported meats. The analytical scheme developed does not require column chromatographic cleanup; chlorfiuazuron was extracted with diethyl ether and petroleum ether (50: 50), partitioned against acetonitrile, cleaned up with silica Sep-Pak cartridge, identified GC/ECD, and comfirmed by GC/MS. The mean recoveries of the pesticide in meat fortified with standard solution 0.1, 0.5, 0.1 mg/kg were ranged from 82 to 95%. The limit of detection and limit of quantitation were 0.001 and 0.005 mg/kg, respectively. Chlorfluazuron residues were not found in domestic samples, but found in imported Australian beef ranging from 0.02 to 0.17 mg/kg, detected by 18% among the samples.

  • PDF

Development of a Chloroform Reference Material for the Proficiency Testing of Hazardous Compounds in Commercial Consumer Chemical Products Under the Consumer Chemical Products and Biocide Safety Management Act (K-BPR)

  • Lee, Sang Tak;Lee, Jae-ung;Kim, Joo-Hyon;Oh, Han Bin
    • Mass Spectrometry Letters
    • /
    • 제10권4호
    • /
    • pp.112-116
    • /
    • 2019
  • In this study, a chloroform consumer chemical product (CCP) reference material (RM) is successfully developed, with potential to be used in the proficiency testing of hazardous compounds in CCPs for analysis and testing agencies. Validation experiments are rigorously conducted to evaluate whether the RM meets the requirements set by the ISO 13528 and ISO Guide 35, using a reliable GC/MS method for the analysis of chloroform. The obtained calibration plot linearity, limit of detection (LOD), and limit of quantitation (LOQ) are excellent. The developed RM meets the homogeneity and stability requirements; the between-unit (sbb) and within-unit (swb) standard deviations are less than 2.5%, and the stability is found to be guaranteed for 50 days.

의약외품중 산화에칠렌가스의 잔류량 분석 (Determination of the Residual Ethylene Oxide in Quasi-drugs)

  • 이정표;김경옥;손경훈;양성준;백옥진
    • 약학회지
    • /
    • 제45권5호
    • /
    • pp.437-441
    • /
    • 2001
  • The quasi-drugs including nonwoven fabric and gauze were sterilized using ethylene oxide (EO) gas. Residual EO in the quasi-drugs was extracted with water (20 mL of water for 1 g of sample) for 24h at 37$^{\circ}C$. Residual EO was determined using GC. The optimal analytical conditions were as follows : column, Carbowax 20M (1.D. 0.2 mm); mobile phase, helium with 30 mL/min; oven temperature 57$^{\circ}C$, injector temperature 18$0^{\circ}C$, detector temperature 20$0^{\circ}C$. The detection limit for EO was 0.10$\mu$g/mL. When the residual EO extracted from nonwoven fabric and gauze was determined, it took more than 9h to get the lower level than 25 ppm which is the limit value of FDA guideline. When the EO residues, ethylene chlorohydrine (ECH) and ethylene glycol (EG) in the 7 commercially available quasi-drugs were determined, no residual EO, ECH, EG were found from the seven commercially available quasi-drugs analyzed by this method.

  • PDF

Studies on the Analysis of Anti-impotent Drugs(II) - Rapid analysis of Sildenafil and modified Sildenafils using HPTLC

  • Choi, Hyun-Cheol;Kang, Sin-Jung;Yun, Mi-Ok;Park, Sang-Aeh;Kim, Ho-Jeong;We, Sae-Seung
    • 대한약학회:학술대회논문집
    • /
    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
    • /
    • pp.396.1-396.1
    • /
    • 2002
  • HPTLC(High Performance Thin Layer Choromatography) method was developed for rapid and precise analysis of Sildenafil and modified Sildenafils(Vardenafil. Homosildenafil, Tadanafil). Chromatographic conditions were Optimized for simultaneous analysis of them and each specific UV spectra were obtained. The calibration curve of Sildenafil and modified Sildenafils had a linearity in the range of 1.0 ~ 56.5 $\mu$/ml at 254nm. The Limit of Detection(LOD) and the limit of Quantification(LOQ) of Sildenafil and modified Sildenafils were 0.8$\mu$/ml and 1.0$\mu$g/ml. (omitted)

  • PDF