• Title/Summary/Keyword: method detection limit

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The Rapid Detection of Pathogens in Organically Grown Vegetables Using PCR-DGGE (PCR-DGGE를 이용한 유기농 채소의 유해 미생물 신속 검지)

  • Kwon, Oh Yeoun;Son, Seok Min
    • Food Engineering Progress
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    • v.15 no.4
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    • pp.370-375
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    • 2011
  • In this study the polymerase chain reaction (PCR) combined with denaturing gradient gel electrophoresis (DGGE) was evaluated as a method permitting the rapid detection of pathogens in fresh originally grown vegetables. A universal primer (341GCf/534r) was selected for its ability to amplify the V3 region of 16S-rRNA genes in their target pathogens (Salmonella typhimurium, Pseudomonas fluorescens, Bacillus cereus, Listeria monoytogenes, Staphyloocus aureus, E. coli). The 194 bp fragments in PCR were successfully duplicated as expected. The amplified fragments of the same size from six different pathogens also showed good separation upon DGGE. The detection limit of PCR-DGGE for six pathogens in fresh-cut lettuces were over $10^{5}$ CFU/g when sampled by stomaching. However, when the sampling method was changed from stomaching to shaking, the detection limit of six pathogens in organic vegetables was shown to increase by over $10^{1}$ CFU/g, but only those of B. cereus were over $10^{3}$ CFU/g. Therefore, PCR-DGGE was shown to be a reliable method for the detection of pathogens in fresh-cut vegetables.

Study on Reduction of Microbial Contamination on Daruma by Combination Treatment of Strong Acidic Hypochlorous Water and Ultrasonic Waves (강산성차아염소산수와 초음파를 병용처리한 조미오징어 반가공품의 미생물 오염도 저감화에 관한 연구)

  • Chung, Won-Hee;Ko, Jun-Soo;Shin, Il-Shik
    • Journal of Food Hygiene and Safety
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    • v.30 no.2
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    • pp.166-172
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    • 2015
  • This study was performed to develop treatment method for reducing microbial contamination on Daruma (a semi-processed product of seasoned and dried squid) by combination of strong acidic hypochlorous water (SAHW) and ultrasonic waves (UW). The available chlorine concentration, oxidation reduction potential (ORP) and pH of SAHW were $69.67{\pm}0.58ppm$, $1071.33{\pm}4.16mV$ and 2.79, respectively. The 1.49 log CFU/g of viable cell count and 1.32 log CFU/g of Staphylococcus aureus was reduced, and Escherichia coli was reduced below detection limit when the Daruma was treated with 20 times (w/v) of sodium hypochlorite solution (SHS) for 120 min. The 3.62 log CFU/g of viable cell count and 3.22 log CFU/g of Staphylococcus aureus was reduced, and Escherichia coli was reduced below detection limit when the Daruma was treated with 20 times (w/v) of SAHW for 120 min. The antibacterial effects of SAHW were stronger than those of SHS at same available chroline concentration. SAHW treatment after washing strongly alkalic electrolyzed water (SAEW) showed better bactericidal effects than SAHW treatment only. The 4.0 log CFU/g of viable cell count was reduced, S. aureus was reduced below regulation limit (Log 2.0 CFU/g), and E. coli was reduced below detection limit when the Daruma was treated with 20 times (w/v) of SAHW for 90 min after washing with 20 times (w/v) of SAEW for 60 min. The viable cell number was reduced below detection limit and S. aureus was reduced below regulation limit when the Daruma was treated with 20 times (w/v) of SAHW for 60 min in ultrasonic washer. E. coli was reduced below detection limit when the Daruma was treated with 20 times (w/v) of SAHW for 10 min in ultrasonic washer. These results suggest that combination of SAHW and UW may be a good technique to reduce the microbial contamination in daruma.

Isolation and HPLC-DAD validation of xanthoangelol in Lespedeza bicolor extract (싸리나무 추출물의 Xanthoangelol 분리 및 HPLC-DAD 밸리데이션)

  • Woo, Hyun Sim;Kim, Yeong-Su;Oh, Yu Jin;Cho, Hae Jin;Song, Se-Kyu;Kim, Dae Wook
    • Korean Journal of Food Science and Technology
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    • v.52 no.1
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    • pp.26-30
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    • 2020
  • This study was undertaken to determine the characteristics of xanthoangelol, the major chalcone constituent derived from the extracts of different parts of Lespedeza bicolor. Xanthoangelol was isolated from the root extract using column chromatography and used as a standard for quantitative analysis. The structure of the isolated compound was established based on spectroscopic evidence. The HPLC-DAD method was validated for specificity, linearity, precision, accuracy, limit of detection, and limit of quantitation. The calibration curve of xanthoangelol had significant linearity (R2>0.9999). Limit of detection and limit of quantitation 0.018 and 0.059 ㎍/mL, respectively. The relative standard deviation values of precision test, and intra- and inter-day tests were less than 0.22 and 0.40%, respectively. In the recovery test, the accuracy ranged from 98.98-102.78% with RSD values less than 0.13%. The method validation parameters indicate the applicability of the HPLC method for quality control of food or drug formulations containing L. bicolor.

Analysis of didecydimethylammonium chloride (DDAC) aerosol in inhalation chamber (흡입챔버 내 didecyldimethylammonium chloride(DDAC) 에어로졸의 분석)

  • Yang, Jeong-Sun;Choi, Sung-Bong;Park, Sang-Yong;Lee, Sung-Bae
    • Analytical Science and Technology
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    • v.25 no.5
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    • pp.307-312
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    • 2012
  • To perform inhalation toxicity test by using experiment animals, we set up an analytical method to monitor didecyldimethylammonium chloride (DDAC) in aerosol nebulized into inhalation chambers by ion chromatography. DDAC was adsorbed by XAD-2 resin and analyzed with conductivity detector. Recovery of DDAC desorbed by acetonitrile from XAD adsorbent was 87.8%. The method detection limit (MDL) and the limit of quantitation (LOQ) were 2.97 ${\mu}g/m^3$ and 8.92 ${\mu}g/m^3$, respectively. Repeatability was calculated as RSD 7.8% in the range of 0~20 ${\mu}g/mL$. Time needed to analyze a sample was less than 5 minutes. Therefore, the analysis of DDAC by ion chromatography was practically useful in monitoring DDAC in inhalation chambers with rapidity and sensitivity manner to perform inhalation toxicity test using experimental animals.

Determination and Validation of Synthetic Antioxidants in Processed Foods Distributed in Korea

  • Park, Hyeon-Ju;Seo, Eunbin;Park, Jin-Wook;Yun, Choong-In;Kim, Young-Jun
    • Journal of Food Hygiene and Safety
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    • v.37 no.5
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    • pp.297-305
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    • 2022
  • Antioxidants are food additives that extend the shelf life of food products by preventing lipid rancidity caused by active oxygen. They can either be naturally-derived or manufactured synthetically via chemical synthesis. In this study, method validation of five synthetic antioxidants, namely butylated hydroxyanisole, butylated hydroxytoluene, tertiary butylhydroquinone, propyl gallate, and disodium ethylenediaminetetraacetic acid, was performed using a high performance liquid chromatography-ultraviolet visible detector, and the method applicability was evaluated by analyzing foods containing antioxidants. The coefficient of determination (R2) average was 0.9997, while the limit of detection and limit of quantification were 0.02-0.53 and 0.07-1.61 mg/kg, respectively. The intra and inter-day accuracies and precisions were 83.2±0.7%-98.7±2.1% and 0.1%-5.7% RSD, respectively. Inter-laboratory validation for accuracy and precision was conducted using the Food Analysis Performance Assessment Scheme quality control material. The results satisfied the guidelines presented by the AOAC International. In addition, the expanded uncertainty was less than 16%, as recommended by CODEX. Consequently, to enhance public health safety, the results of this study can be used as basis data for evaluating the intake of synthetic antioxidants and assessing their risks in Korea.

Analysis of germanium in rock and sediment by ICP/MS after ammonium bifluoride(NH4HF2) digestion (이플루오린화 암모늄 시료분해 및 ICP/MS에 의한 암석 및 퇴적물 중 게르마늄 분석)

  • Eum, Chul Hun;Choi, Won Myung
    • Analytical Science and Technology
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    • v.26 no.6
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    • pp.375-380
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    • 2013
  • Ammonium biflouride ($NH_4HF_2$) digestion was studied for germanium analysis in rock and sediment by inductively coupled plasma mass spectrometry (ICP/MS). QLO-1 and SDO-1 are used for reference materials from USGS. Sediment, basalt and ball clay for GeoPT were chosen as real samples. The loss of germanium in open vessel digestion was well known which can be caused by easy transformation to volatile compounds. But ammonium bifluoride digestion could suppress loss of germanium in open vessel digestion. Germanium recovery was not influenced by hydrogen peroxide with ammonium bifluoride digestion. Furthermore, the new method was simple and rapid in germanium analysis by ICP/MS. MDL(method detection limit) was 0.015 ${\mu}g/g$ and germanium recovery was 106~128%.

An Alternative Method for Extracting Plasmodium DNA from EDTA Whole Blood for Malaria Diagnosis

  • Seesui, Krongkaew;Imtawil, Kanokwan;Chanetmahun, Phimphakon;Laummaunwai, Porntip;Boonmars, Thidarut
    • Parasites, Hosts and Diseases
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    • v.56 no.1
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    • pp.25-32
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    • 2018
  • Molecular techniques have been introduced for malaria diagnosis because they offer greater sensitivity and specificity than microscopic examinations. Therefore, DNA isolation methods have been developed for easy preparation and cost effectiveness. The present study described a simple protocol for Plasmodium DNA isolation from EDTA-whole blood. This study demonstrated that after heating infected blood samples with Tris-EDTA buffer and proteinase K solution, without isolation and purification steps, the supernatant can be used as a DNA template for amplification by PCR. The sensitivity of the extracted DNA of Plasmodium falciparum and Plasmodium vivax was separately analyzed by both PCR and semi-nested PCR (Sn-PCR). The results revealed that for PCR the limit of detection was $40parasites/{\mu}l$ for P. falciparum and $35.2parasites/{\mu}l$ for P. vivax, whereas for Sn-PCR the limit of detection was $1.6parasites/{\mu}l$ for P. falciparum and $1.4parasites/{\mu}l$ for P. vivax. This new method was then verified by DNA extraction of whole blood from 11 asymptomatic Myanmar migrant workers and analyzed by Sn-PCR. The results revealed that DNA can be extracted from all samples, and there were 2 positive samples for Plasmodium (P. falciparum and P. vivax). Therefore, the protocol can be an alternative method for DNA extraction in laboratories with limited resources and a lack of trained technicians for malaria diagnosis. In addition, this protocol can be applied for subclinical cases, and this will be helpful for epidemiology and control.

Simultaneous Determination of Six Components in the Traditional Herbal Medicine 'Oryeongsan' by HPLC-DAD and LC-MS/MS

  • Lee, Jiwoo;Weon, Jin Bae;Lee, Bohyoung;Yun, Bo-Ra;Eom, Min Rye;Ma, Choong Je
    • Natural Product Sciences
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    • v.19 no.1
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    • pp.20-27
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    • 2013
  • A simple high performance liquid chromatography - diode array detector (HPLC-DAD) method has been developed and validated for simultaneous determination of the six components (cinnamic acid, cinnamaldehyde, eugenol, atractylenolide I, atractylenolide III, and ergosterol) in Oryeongsan. In addition, identification of six marker compounds was conducted by a LC-MS/MS analysis. The six compounds in Oryeongsan were separated on Shishedo $C_{18}$ column (5 ${\mu}m$, $4.6{\times}250$ mm) at a column temperature of $30^{\circ}C$. The mobile phase was a mixture of 0.1% trifluoroacetic acid (TFA) water and acetonitrile employing gradient elution at a flow rate of 1.0 mL/min. The detection wavelength was set at 205, 250, 280, and 330 nm. The developed method had good linearity ($R^2$ > 0.9997) and the limit of detection (LOD) and limit of quantification (LOQ) were observed within the ranges 0.01~0.15 and 0.05~0.45 ${\mu}g/mL$, respectively. The relative standard deviation (RSD) values of intra- and inter-day testing were indicated that less than 3% and 90.31~103.31% of accuracy. The results of recovery test were 90.56~106.72% and RSD range was measured from 0.84 to 2.95%. In conclusion, this HPLC-DAD method has been successfully applied to the simultaneous determination of six compounds in Oryeongsan samples.

Marker compounds contents of Salvia miltiorrhiza Radix depending on the cultivation regions

  • Seong, Gi-Un;Kim, Mi-Yeon;Chung, Shin-Kyo
    • Journal of Applied Biological Chemistry
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    • v.62 no.2
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    • pp.129-135
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    • 2019
  • Salvia miltiorrhiza Radix is cultivated in Korea and China and is traditionally used to treat cardiovascular diseases. In this study, we developed and validated a quantitative analysis method for S. miltiorrhiza Radix using high-performance liquid chromatography (HPLC). Identification was performed using ultra performance liquid chromatography-tandem mass spectrometry. For quantitative analysis, we used seven marker compounds. Separation conditions for HPLC were optimized using an ODS column with gradient conditions of 1% formic acid in distilled water and 1% formic acid in acetonitrile, with a flow rate of 0.8 mL/min and a detection wavelength of 280 nm. This method showed good linearity ($R^2=0.9998$), precision (relative standard deviation ${\leq}3.3%$), accuracy (recovery of 94.16-102.89%), limit of detection ($7.53{\mu}g/mL$), and limit of quantification ($23.71{\mu}g/mL$). This approach successfully quantified marker compounds in S. miltiorrhiza Radix. The individual marker compounds were identified by comparing the molecular masses and retention times with does standard compounds. Marker compound contents of S. miltiorrhiza Radix were investigated with different cultivation regions. Seven marker compounds were detected and quantified in all samples. Among them, salvianolic acid B showed the highest contents and it ranged from 4.13 to 7.15%. The salvianolic acid B content (7.15%) of marker compound was the highest in Bonghwa, and the tanshinone IIA content (1.90%) was the highest in Pohang. The results of marker compounds and developed method were intended to provide a favorable reference for the study of S. miltiorrhiza Radix from different regions of Korea.

Raman spectroscopic analysis to detect olive oil mixtures in argan oil

  • Joshi, Rahul;Cho, Byoung-Kwan;Joshi, Ritu;Lohumi, Santosh;Faqeerzada, Mohammad Akbar;Amanah, Hanim Z;Lee, Jayoung;Mo, Changyeun;Lee, Hoonsoo
    • Korean Journal of Agricultural Science
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    • v.46 no.1
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    • pp.183-194
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    • 2019
  • Adulteration of argan oil with some other cheaper oils with similar chemical compositions has resulted in increasing demands for authenticity assurance and quality control. Fast and simple analytical techniques are thus needed for authenticity analysis of high-priced argan oil. Raman spectroscopy is a potent technique and has been extensively used for quality control and safety determination for food products In this study, Raman spectroscopy in combination with a net analyte signal (NAS)-based methodology, i.e., hybrid linear analysis method developed by Goicoechea and Olivieri in 1999 (HLA/GO), was used to predict the different concentrations of olive oil (0 - 20%) added to argan oil. Raman spectra of 90 samples were collected in a spectral range of $400-400cm^{-1}$, and calibration and validation sets were designed to evaluate the performance of the multivariate method. The results revealed a high coefficient of determination ($R^2$) value of 0.98 and a low root-mean-square error (RMSE) value of 0.41% for the calibration set, and an $R^2$ of 0.97 and RMSE of 0.36% for the validation set. Additionally, the figures of merit such as sensitivity, selectivity, limit of detection, and limit of quantification were used for further validation. The high $R^2$ and low RMSE values validate the detection ability and accuracy of the developed method and demonstrate its potential for quantitative determination of oil adulteration.