• Title/Summary/Keyword: Reaction monitoring

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Review and application of environmental DNA (eDNA) investigation of terrestrial species in urban ecosystem (도시 내 육상 생물종 모니터링을 위한 환경DNA 리뷰 및 적용)

  • Kim, Whee-Moon;Kim, Seoung-Yeal;Park, Il-Su;Lee, Hyun-Jung;Kim, Kyeong-Tae;Kim, Young;Kim, Hye-Joung;Kwak, Min-Ho;Lim, Tae-Yang;Park, Chan;Song, Won-Kyong
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.23 no.2
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    • pp.69-89
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    • 2020
  • Scientific trust and quantification of traditional species investigation and results that have been used in ecology for decades has always been a problem and concern for ecologists. Global ecologists have proposed DNA-based species investigation studies to find answers to problems. In this study, we reviewed the global trend of research on environmental DNA(eDNA), which is a method for monitoring species by detecting DNA of organisms naturally mixed in environmental samples such as water, soil, and feces. The first eDNA research confirmed the possibility of species investigation at the molecular level, and commercialization of NGS(Next Generation Sequencing) and DNA metabarcoding elicits efficient and quantitative species investigation results, and eDNA research is increasing in the filed of ecology. In this study, mammals and birds were detected using MiMammal universal primers from 23 samples(3 natural reserves; 20 water bowls) out of 4 patches to verify eDNA for urban ecosystems in Suwon, and eDNA was verified by performing camera trapping and field survey. Most terrestrial species were detected through eDNA, and particularly, mice(Mus musculus), and Vinous-throated Parrotbill (Sinosuthora webbiana) were identified only with eDNA, It has been confirmed to be highly effective by investigating techniques for small and internal species. However, due to the lack of resolution of the primer, weasels(Mustela sibirica) and squirrels(Melanochromis auratus) were not detected, and it was confirmed that the traditional investigation method was effective only for a few species, such as Mogera robusta(Mogera robusta). Therefore, it is judged that the effects of species investigation can be maximized only when eDNA is combined with traditional field survey and Camera trapping to complement each other.

Development of LC-MS/MS Quantitation Method for Ethoxyquin in Fishery Products (수산물 중 에톡시퀸의 LC-MS/MS 정량분석법 개발)

  • Shin, Dasom;Chae, Young-Sik;Kang, Hui-Seung;Lee, Soo-Bin;Cho, Yoon-Jae;Cheon, So-Young;Jeong, Jiyoon;Rhee, Gyu-Seek
    • Journal of Food Hygiene and Safety
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    • v.31 no.6
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    • pp.432-438
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    • 2016
  • Ethoxyquin (EQ, 1,2-dihydro-6-ethoxy-2,2,4-trimethyl-quinoline) is quinoline-based antioxidant used in the animal feed and food industry to protect the raw materials and final products against oxidation. In recent years the use of synthetic antioxidants in fishmeal ingredients carry-over to farmed fish fillets has received increasing attention in food safety. This study was conducted to develop an analytical method to determine EQ in aquatic products. The analytes were confirmed and quantified via liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the positive ion mode using multiple reaction monitoring (MRM). The sample was extracted with 1 N HCl (in case of flatfish extracted with 1 N HCl containing 10% acetonitrile). Then, solid phase extraction (SPE) was used for the cleanup. Standard calibration curves presented linearity with the correlation coefficient ($r^2$) > 0.99, analyzed at 0.005-0.2 mg/kg concentration. The developed method was validated according to the Codex Alimentarius Commission (CAC) guideline. The limits of quantitation for EQ were 0.01 mg/kg. Average recoveries ranged from 81.3% to 107%. The repeatability of measurements, expressed as the coefficient of variation (CV, %), was below 10%. The analytical method was characterized with high accuracy and acceptable sensitivity to meet CODEX guideline requirements and would be applicable to analyze the EQ residue in aquatic products.

Development of Analytical Method for Determination of Baicalin, Eleutheroside E, and Ligustilide by LC-MS/MS in Raw Materials of Health Functional Foods (LC-MS/MS를 이용한 건강기능식품 원료 중 바이칼린, 엘레우테로사이드 E, 리구스틸라이드 동시분석법 개발)

  • Keum, Eun Hee;Chung, So Young;Lee, Jin Hee;Kim, Meehye
    • Journal of Food Hygiene and Safety
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    • v.32 no.3
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    • pp.193-198
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    • 2017
  • Scutellaria baicalensis, Eleutherococcus senticosus, and Angelica sinensis have been used as raw materials for health functional foods. This study was conducted to develop a novel method to analyze levels of baicalin (Scutellaria baicalensis), eleutheroside E (Eleutherococcus senticosus), and ligustilide (Angelica sinensis) simultaneously in health functional foods. The methanol extracted samples were analyzed and quantified via liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the positive ion mode and the negative ion mode using multiple reaction monitoring. Standard calibration curves confirmed linearity with the correlation coefficient ($r^2$) of > 0.99 at $100-2000{\mu}g/mL$ concentration range. The limits of detection (LODs) and limits of quantitation (LOQs) were in the range of $13.0{\sim}35.2{\mu}g/L$ and $39.3{\sim}106.7{\mu}g/L$, respectively. The recovery results ranged between 91.4~109.9% at 3 different concentration levels with relative standard deviations (RSDs) less than 5%. The proposed analytical method was characterized with high accuracy and acceptable precision. The new method would be an effective tool to analyze baicalin, eleutheroside E, and ligustilide simultaneously in raw materials of health functional foods.

Monitoring of Chicken RNA Integrity as a Function of Prolonged Postmortem Duration

  • Malila, Yuwares;Srimarut, Yanee;U-chupaj, Juthawut;Strasburg, Gale;Visessanguan, Wonnop
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1649-1656
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    • 2015
  • Gene expression profiling has offered new insights into postmortem molecular changes associated with meat quality. To acquire reliable transcript quantification, high quality RNA is required. The objective of this study was to analyze integrity of RNA isolated from chicken skeletal muscle (pectoralis major) and its capability of serving as the template in quantitative real-time polymerase chain reaction (qPCR) as a function of postmortem intervals representing the end-points of evisceration, carcass chilling and aging stages in chicken abattoirs. Chicken breast muscle was dissected from the carcasses (n = 6) immediately after evisceration, and one-third of each sample was instantly snap-frozen and labeled as 20 min postmortem. The remaining muscle was stored on ice until the next rounds of sample collection (1.5 h and 6 h postmortem). The delayed postmortem duration did not significantly affect $A_{260}/A_{280}$ and $A_{260}/A_{230}$ ($p{\geq}0.05$), suggesting no altered purity of total RNA. Apart from a slight decrease in the 28s:18s ribosomal RNA ratio in 1.5 h samples (p<0.05), the value was not statistically different between 20 min and 6 h samples ($p{\geq}0.05$), indicating intact total RNA up to 6 h. Abundance of reference genes encoding beta-actin (ACTB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase (HPRT), peptidylprolylisomerase A (PPIA) and TATA box-binding protein (TBP) as well as meat-quality associated genes (insulin-like growth factor 1 (IGF1), pyruvate dehydrogenase kinase isozyme 4 (PDK4), and peroxisome proliferator-activated receptor delta (PPARD) were investigated using qPCR. Transcript abundances of ACTB, GAPDH, HPRT, and PPIA were significantly different among all postmortem time points (p<0.05). Transcript levels of PDK4 and PPARD were significantly reduced in the 6 h samples (p<0.05). The findings suggest an adverse effect of a prolonged postmortem duration on reliability of transcript quantification in chicken skeletal muscle. For the best RNA quality, chicken skeletal muscle should be immediately collected after evisceration or within 20 min postmortem, and rapidly preserved by deep freezing.

Objective Identification of Human Sperm Hyperactivation by Computerized Sperm Motion Analysis (Computerized Sperm Motility Analyzer를 이용한 Human Sperm의 Hyperactivated Motility의 객관적 관찰에 관한 연구)

  • Lee, Hee-Kyung;Lee, Chan;Kim, Hyun-Sook;Kim, Young-Tae;Kim, Sun-Haeng;Ku, Pyoung-Sahm
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.1
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    • pp.1-11
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    • 1994
  • The occurrence and time course of capacitation, acrosomal loss, and hyperactivated motility require quantitative definition in order to characterize fertile human sperm. Recently the method has been developed to estimate the quality of spermatozoa by using kinematic parameters such as curvilinear velocity(VCL), average path velocity(VAP), linearity(LIN), straightness(STR), amplitude of lateral head displacement(ALH), and beat cross frequence(BCF) from Computer Assisted Sperm Analysis (CASA). In this study, using the Hamilton Thorn motility analyzer HTM 2030(Hamilton Thorn Research, Beverly, MA), we attempted to identify the spermatozoa with hyperactivated motility (HA) objectively and to monitor hyperactivation of human spermatozoa during incubation in capacitating media and after treatment of calcium ionophore as compared with acrosome status. And we examined whether HA are related to the result of SPA. Semen samples obtained from 16 healthy men were prepared by swim up technique and preincubated in a capacitating media(modified BWW medium) for 5 hours and treated with calcium ionophore solution. The acrosome reaction was detected with PSA-FITC labelling of the acrosome and in vitro sperm ferilizing capacity was assessed by the zona free hamster ovum penetration assay (SPA). The incidence of hyperactivated sperm was 2.6% in fresh semen, 14.3% of the swim up population, 13.7% after 5h of incubation. Significant increase of percentage of hyperactivated sperm was observed after the incubation (p<0.05) but after treatment, no significant changes of percentage of hyperactivated sperm(l1.8%) in contrast to significant rise in the percentage of acrosome reacted cells. Correlation analysis failed to show any significant relationship between the percentage of sperm with HA and SPA score. In conclusion, although no direct correlations were found between the results of SPA and HA, hyperactivation of sperm is associated with capacitation and monitoring hyperactivated sperm will be expected as a method of evaluating the functional quality of sperm such as SPA.

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Validation of an LC/MS/MS Method for the Pharmacokinetic Study of Lercanidipine (염산레르카니디핀 체내동태 연구를 위한 혈청 중 레르카니디핀의 LC/MS/MS 정량법 검증)

  • Kim, Se-Mi;Kim, Hwan-Ho;Shin, Sae-Byeok;Kang, Hyun-Ah;Yoon, Hwa;Cho, Hea-Young;Kim, Yoon-Gyoon;Yang, Chan-Woo;Yong, Chul-Soon;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.37 no.4
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    • pp.223-227
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    • 2007
  • A rapid, simple and sensitive LC/MS/MS method for the determination of lercanidipine in human serum was validated and applied to the pharmacokinetic study of lercanidipine. Lercanidipine and internal standard, amlodipine, were extracted from human serum by liquid-liquid extraction with hexan-isoamyl alcohol (100: 1, v/v) and analyzed on a $Symmetry^{(R)}$ MS $C_{18}$ column with the mobile phase of acetonitrile-0.2% aqueous formic acid (70: 30, v/v). Using MS/MS with multiple reaction monitoring (MRM) mode, lercanidipine and amlodipine were detected without severe interferences from human serum matrix. Lercanidipine produced a protonated precursor ion ($[M+H]^+$) at m/z 612.3 and a corresponding product ion at m/z 280.0. Internal standard produced a protonated precursor ion ($[M+H]^+$]) at m/z 409.0 and a corresponding product ion at m/z 238.0. The ruggedness of this method was investigated using quality control (QC) samples. This method showed linear response over the concentration range of 0.05-20 ng/mL with correlation coefficient greater than 0.999. The lower limit of quantitation using 0.5 mL of serum was 0.05 ng/mL, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the developed method ranged from 85.51 to 112.2% for lercanidipine with overall precision (% C.V.) being 3.56-13.1%. This method showed good ruggedness (within 15% C.V.) and was successfully applied for the analysis of lercanidipine in human serum samples for the pharmacokinetic studies, demonstrating the suitability of the method.

The Effects of Scale Growth Inhibition on Water Pipe using Frequency Driver (Frequency Driver를 이용한 냉온수관의 스케일 방지억제효과)

  • Jang, Mi-Jeong;Sung, Il-Wha
    • Journal of Korean Society of Environmental Engineers
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    • v.33 no.4
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    • pp.258-266
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    • 2011
  • Scale induced by hardness materials in water must be controled because of it can be result in remarkable damages of pipeline as well as water quality deterioration. Especially hot water system is one of scale management required facility as scale formation can be accelerated by temperature. The scale control performance of frequency driver (FD) was tested instead of existing methods such as chemical, physical and electromagnetic methods which needs chemicals and electric power. Three kinds of pipe coupons were submerged in test water with 500 mg/L of hardness for 33 days and XRD and SEM were analysed for comparing scale formation characteristics of these coupons. Calcite ($CaCO_3$) which came from hardness of water was formed on only cast iron pipe coupon and this coupon showed higher corrosion rate than copper and stainless steel pipe coupon. Hot water circulating system connected cast iron pipe with and without FD was operated with 300 mg/L of hardness water at $50^{\circ}C$ for monitoring of scale formation and water quality with and without FD. XRD showed that FD leaded to magnetite ($Fe_3O_4$) scale which is good scale for preventing corrosion than calcite and SEM image also indicated the scale control effect of FD. Scales of 16% on pipe joint, 14% on pipe length, and 42% on heat exchanger decreased with FD comparing scales of those parts without FD. From the results of water quality, FD reduced crystallization of hardness material without chemical reaction in water and it can indicate that FD is safe and proenvironmental technology for scale reduction.

Lifestyle intervention might easily improve blood pressure in hypertensive men with the C genotype of angiotensin II type 2 receptor gene

  • Kitaoka, Kaori;Kitade, Azusa;Nagaoka, Junko;Tsuzaki, Kokoro;Harada, Kiyomi;Aoi, Wataru;Wada, Sayori;Asano, Hiroaki;Sakane, Naoki;Higashi, Akane
    • Nutrition Research and Practice
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    • v.9 no.4
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    • pp.385-392
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    • 2015
  • BACKGROUND/OBJECTIVES: Recent studies have reported an association of the angiotensin II type 2 receptor (AT2R) 3123Cytosine/Adenine (3123C/A) polymorphism with essential hypertension and cardiovascular diseases. The purpose of the study was to investigate whether the AT2R 3123C/A polymorphism affects blood pressure for free-living hypertensive men during a 5-month intervention period. SUBJECTS/METHODS: The subjects were free-living hypertensive Japanese men aged 40 to 75 years who agreed to intervention in the period from 2004 to 2011. Detection of the AT2R 3123C/A polymorphism was determined by polymerase chain reaction. The dietary intervention was designed to decrease salt level and to increase potassium level through cooking instructions and self-monitoring of the diet. The exercise session consisted of activities such as stretching, resistance training, and walking. Blood pressure, urinary sodium and potassium excretion, dietary and lifestyle data, and non-fasting venous blood sample were collected at baseline and after the intervention period. RESULTS: Thirty nine subjects were eligible for participation and the follow-up rate was 97.4%. The C allele proportion was 57.9%. AT2R 3123C/A polymorphism was X-chromosome-linked, therefore we analyzed the C and A genotypes. At baseline, no significant differences were observed between the genotype groups. After the intervention, there were no significant differences in lifestyle habit between the groups. Nevertheless, the estimated salt excretion (g/day) was significantly decreased only in the C genotype (13.0-10.3, P = 0.031). No significant change was observed in systolic blood pressure (SBP) (mmHg) in the A genotype, but a significant decrease was observed in the C genotype (150.0-141.5, P = 0.024). CONCLUSTIONS: In the C genotype, it might be easy to improve SBP through lifestyle intervention in free-living hypertensive Japanese men, however generalization could not be achieved by the small sample size.

Survey and method validation of simultaneous quantitative analysis of T-2 and HT-2 toxins in cereals (곡류 중 T-2 및 HT-2 독소 동시 정량분석의 유효성 검증 및 실태조사)

  • Paek, Ockjin;Kang, Teabeom
    • Food Science and Preservation
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    • v.22 no.4
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    • pp.559-566
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    • 2015
  • The aim of this study was to develop an analytical method for determination of T-2 toxin and HT-2 toxin level in cereals and to survey their levels using LC-MS/MS. The T-2 and HT-2 toxins were simultaneously analyzed by electrospray ionization with a positive ion mode and multiple reaction monitoring (MRM) after filteration and immuno-affinity column clean-up. A matrix-matched standard calibration used for quantification and recoveries of T-2 and HT-3 toxins were in the range of $100.6{\pm}7.2%$ and $96.8{\pm}9.4%$, respectively. Limits of detection and quantification of T-2 and HT-2 toxins were estimated to be 0.5 and $1.5{\mu}g/kg$, respectively. Each repeatability (RSRr) of T-2 and HT-2 toxins was determined to be 0.9~6.0%, and 4.9~6.1%, respectively. Total 115 samples cereals were collected from 9 types of cereals for analysis. The positive percentages of T-2 and HT-2 toxins obtained from collected samples were found to be 72% and 80%, respectively. The contamination level of T-2 toxin and HT-2 toxin in cereals were $37.1{\mu}g/kg$, and $5.4{\mu}g/kg$, respectively. Therefore, this study suggests that the developed method could be an useful analytical method to determine the T-2 and HT-2 toxin level in cereals and the present data could be used as a reference to estimate the risk assessment.

Thick Film Gas Sensor Based on PCB by Using Nano Particles (나노 입자를 이용한 PCB 기반 후막 가스 센서)

  • Park, Sung-Ho;Lee, Chung-Il;Song, Soon-Ho;Kim, Yong-Jun
    • Journal of the Microelectronics and Packaging Society
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    • v.14 no.2 s.43
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    • pp.59-63
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    • 2007
  • This paper presented a low-cost thick film gas sensor module, which was based on simple PCB (Printed Circuit Board) process. The proposed sensor module included a $NO_2/H_2$ gas sensor, a relative humidity sensor, and a heating element. The $NO_2/H_2$ gas and relative humidity sensors were realized by screen-printing $SnO_2,\;BaTiO_3$ nano-powders on IDTS (Interdigital Transducer) of a PCB substrate, respectively. At first 1% $H_2$ gas flowed into the sensor chamber. After 4 min, air filled the chamber while $H_2$ gas flow stopped. This experiment was performed repeatedly. The Identical procedure was used for the $NO_2$ detection. The result for sensing $H_2$ gas showed the increase of voltage from 0.8V to 3.5V due to the conductance increase and its reaction response time by hydrogen flow was 65 sec. $NO_2$ sensing results showed 2.7 V voltage drop due to the conductance decrease and its response time was 3 sec through a voltage monitoring.

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