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Simultaneous Determination of Non-steroidal Anti-inflammatory Drugs and Corticosteroids Added to Foods as Adulterants using LC-ESI-tandem Mass Spectrometry (LC/ESI-MS/MS를 이용한 식품 중 불법적으로 첨가된 비스테로이드성 소염진통제 및 스테로이드 의약품 동시분석)

  • Lee, Yongcheol;Park, Ju-Sung;Kim, Sung-Dan;Yang, Hye-Ran;Kim, Eun-Hee;Yi, Yun-Jung;Cho, Sung-Ja;Jo, Han-Bin;Kim, Jung-Hun;Chae, Young-Zoo
    • Journal of Food Hygiene and Safety
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    • v.28 no.3
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    • pp.247-251
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    • 2013
  • The objective of present study was to develop a simultaneous determination method of 5 medical compounds, including beclomethasone, dexamethasone, prednisolone, ketoprofen, phenylbutazone in foods, using LC-MS/MS. To optimize MS analytical condition of 5 compounds, each parameter was established by MRM mode. The chromatographic separation was achieved on a C18 column successfully, with a mobile phase made up of A (0.1% formic acid) and B (0.1% formic acid in acetonitrile), at a flow rate of 0.3 mL/min for 17 min with a gradient elution. LOD and LOQ of 5 compounds were in the range of 0.40~4.60 ng/mL and 0.81~11.46 ng/mL, respectively. As a result of analyzing the three concentrations of the standard mixture added to blank samples, the results showed that the mean recovery rate of 5 compounds was in the range of 81.52~103.83%, and RSD (%) of Intra- and Inter-day assay were 0.52-10.45. Since relatively fine selectivity, accuracy and reproducibility were shown in this qualified experimental method, it could be utilized efficiently to investigating those 5 compounds to see if it is added to food products illegally.

Water Quality Improvement of Stagnant Water using an Upflow Activated Carbon Biofilm Process and Microbial Community Analysis (상향류 활성탄 생물막 공정을 이용한 정체 수역 수질 개선 및 공정 내 미생물 군집 해석)

  • Oh, Yu-Mi;Lee, Jae-Ho;Park, Jeung-Jin;Choi, Gi-Choong;Park, Tae-Joo;Lee, Tae-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.1
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    • pp.23-32
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    • 2010
  • The capacity of natural purification was limited by the interruption of natural flow and the problems such as eutrophication were occurred by nutritive salts accumulation in stagnant stream. Moreover, the inflow of non-point sources causes non-degradable materials to increase in stagnant stream. In this study, an upflow biological activated carbon (BAC) biofilm process comprised of anoxic, aerobic 1, and aerobic 2 reactors were introduced for treatment of stagnant stream and SS, $BOD_5$, $COD_{Mn}$, $COD_{Cr}$, TN, and TP were monitored in the upflow BAC biofilm reactors with continuous cycling. In order to simulate stagnant stream, the lake water of amusement park and golf course were stored as influent in a tank of $2m^3$ and hydraulic retention time (HRT) was changed into 6, 4, and 2 hours. At HRT 4hr and the lake water of amusement park as influent, the removal efficiencies of SS, $BOD_5$, $COD_{Mn}$, $COD_{Cr}$, TN, and TP showed the best water quality improvement and were 69.8, 83.0, 91.3, 74.1, 74.7, and 88.9%, respectively. At HRT 4hr and the lake water of golf course as influent, the removal efficiencies of SS, $BOD_5$, $COD_{Mn}$, $COD_{Cr}$, TN and TP were 78.5, 78.0, 80.2, 74.9, 55.6 and 97.5%, respectively. As the results of polymerase chain reaction - denaturing gel gradient electrophoresis (PCR-DGGE), microbial community was different depending on influent type. Fluorescence in situ hybridization (FISH) showed that nitrifying bacteria was dominant at HRT 4 hr. The biomass amount and microbial activities by INT-DHA test were not decrease even at lower HRT condition. In this study, the upflow BAC biofilm process would be considered to the water quality improvement of stagnant stream.

Method for Validation of Caffeoylquinic Acid Derivatives in Ligularia fischeri Leaf Extract as Functional Ingredients (건강기능식품 기능성 원료로서 곰취잎 추출물의 Caffeoylquinic Acid계 성분 분석법 검증)

  • Kwon, Jin Gwan;Kim, Jin Kyu;Seo, Changon;Hong, Seong Su;Ahn, Eun-Kyung;Seo, Dong-Wan;Oh, Joa Sub
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.1
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    • pp.61-67
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    • 2016
  • An HPLC analysis method was developed for standard determinations of chlorogenic acid, 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, and 4,5-di-O-caffeoylquinic acid as functional health materials in Ligularia fischeri extract. HPLC was performed on a $C_{18}$ Kromasil column ($4.6{\times}250mm$, $5{\mu}m$ column) with a gradient elution of 0.1% (v/v) trifluoroacetic acid and acetonitrile at a flow rate of 1.0 mL/min at $30^{\circ}C$. The analytes were detected at 330 nm. The HPLC method was validated in accordance with the International Conference on Harmonization guideline of analytical procedures with respect to specificity, precision, accuracy, and linearity. The limits of detection and quantitation for the four compounds were 3.0~14.6 and $9.2{\sim}44.4{\mu}g/mL$, respectively. Calibration curves showed good linearity ($r^2$ > 0.999), and the precision of analysis was satisfied (less than 0.9%). Recoveries of quantified compounds ranged from 98.96 to 101.81%. This result indicates that the established HPLC method is very useful for the determination of marker compounds in Ligularia fischeri leaf extracts.

Simultaneous HPLC Determination of Marker Compounds for the Standardization of Hedyotis diffusa (백운풀의 지표성분 설정 및 품질표준화를 위한 정량 분석법)

  • Bang, Han-Yeol;Yang, Eun-Ju;Kim, Jeong-Ah;Song, Kyung-Sik
    • Journal of Life Science
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    • v.23 no.8
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    • pp.1025-1031
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    • 2013
  • From a 95% ethanolic extract of H. diffusa, four marker compounds (HD1~HD4) were isolated, which were relatively unique and exist in comparably high contents. The structures of marker compounds were identified as digitolutein (1), 2-hydroxy-3-methylanthraquinone (2), (E/Z)-6-O-p-coumaroyl scandoside methyl ester (4:1 mixture) (3), and (E/Z)-6-O-p-methoxycinnamoyl scandoside methyl ester (4:1 mixture) (4), respectively, on the basis of $^{13}C$ and $^1H$-NMR analyses. The calibration curves of marker compounds showed high linearity, as their correlation coefficient ($R^2$) were in the range of 0.9991~0.9999. In addition, the limit of detection (LOD) and the limit of quantification (LOQ) were $0.03{\sim}0.07{\mu}g/ml$ and $0.099{\sim}0.231{\mu}g/ml$, respectively. The intra-day/inter-day precision and accuracy were 0.23~2.00%/0.25~1.16% and 94.60~108.44%/94.73-110.23%, respectively. The optimal HPLC conditions for the simultaneous quantification of HD1~HD4 were as follows: stationary phase; Merck Chromolith RP-18e ($100{\times}4.6mm$, $5{\mu}m$), column temp.; room temperature, UV detection at 280 nm, flow rate; 2.0 ml/min, injection volume; $10{\mu}l$, mobile phase; start with the mixture of 80% solvent A ($H_2O$ containing 0.5% acetic acid) and 20% solvent B (methanol containing 0.5% acetic acid) and gradually decrease solvent A to 40% in 9 min., then retain this condition to 18 min. Under the HPLC condition, the four marker compounds 1~4 were successfully separated without any interference of other constituents. The results obtained in this study are expected to be helpful for the development of nutraceutics and natural medicines and for the quality control of this plant.

Monitoring of Forbidden Medicines as Adulterants in Dietary Supplements Marketed Online by HPLC and ESI-tandom Mass Spectrometry (HPLC 및 ESI-tandom Mass Spectrometry를 이용한 인터넷에서 판매되는 기능성표방식품 중 부정유해물질 모니터링)

  • Lee, Yong-Cheol;Kim, Bog-Soon;Yun, Eun-Sun;Kim, Sung-Dan;Chang, Min-Su;Park, Young-Ae;Shin, Young;Jung, Sun-Ok;Lee, Jae-In;Chae, Young-Zoo
    • Korean Journal of Food Science and Technology
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    • v.44 no.2
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    • pp.148-154
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    • 2012
  • It was carried out to detect anti-impotence drug-like compounds, sibutramine and their analogues in dietary supplements, which are doubtful whether they include illegal compounds. A total of 51 food products were bought online and have been investigated. The separation was achieved on a C18 column, with the mobile phase made up of water (5 mM sodium hexanesulfonate and 0.1% phosphoric acid) and 95% acetonitrile, at a flow rate of 1.2 mL/min with gradient elution using by HPLC-DAD. The UV signals were monitored at 220 nm and 291 nm. LC-ESI-tandom MS was utilized to confirm that detected compounds in samples are the same as the reference materials. Two nutrient supplement foods and ginseng products were found to contain 1.3-82.1 mg of sildenafil, dimethylthiolsildenafil and pseudovardenafil per serving size. In addition, two other processed products were detected to contain 1.7 and 2.2 mg of didesmethylsibutramine, derived from sibutramine per serving size.

Optimization of HPLC Method and Clean-up Process for Simultaneous and Systematic Analysis of Synthetic Color Additives in Foods (식품 중 타르색소의 동시분석 및 계통분석을 위한 HPLC 분석조건 및 정제과정 확립)

  • Park, Sung-Kwan;Hong, Yeun;Jung, Yong-Hyun;Lee, Chang-Hee;Yoon, Hae-Jung;Kim, So-Hee;Lee, Jong-Ok
    • Korean Journal of Food Science and Technology
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    • v.33 no.1
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    • pp.33-39
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    • 2001
  • To develop a method for separation process using Sep-pak $C_18$, simultaneous and systematic analysis of 8 permitted and 11 non-permitted synthetic food colors in Korea, optimization of analysis conditions for reverse phase ion-pair high performance liquid chromatography was carried out. For the best result of Sep-pak $C_18$ separation the pH of color standard mixture solution was $5{\sim}6$ and 0.1% HCl-methanol solution were set as eluent. The colors eluated from Sep-pak $C_18$ cartridge were determined and confirmed by high performance liquid chromatography with a photodiode array detector at 420 nm for yellow colors type, at 520 nm for red colors type, at 600 nm for blue and green colors type and at 254 nm for mixed colors. Conditions for HPLC analysis were as follows: column, Symmetry $C_18$ (5 m, 3.9 mm $i.d.{\times}150\;mm$); mobile phase, 0.025 M ammonium acetate (containing 0.01 M tetrabutylammonium bromide) : acetonitrile : methanol (65 : 25 : 10) and 0.025 M ammonium acetate(containing 0.01 M tetrabutylammonium bromide) : acetonitrile : methanol (40 : 50 : 10); flow rate, 1 mL/min. It takes 35 minutes for simultaneaus analysis and 18 minutes for systematic analysis. The detection limits range of each colors were $0.01{\sim}0.05\;{\mu}g/g$.

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Comparison of Disk Tension Infiltrometer and van Genuchten-Mualem Model on Estimation of Unsaturated Hydraulic Conductivity (장력 침투계(Disk Tension Infiltrometer)와 van Genuchten-Mualem 모형 적용에 따른 불포화수리 전도도의 비교 해석)

  • Hur, Seung-Oh;Jung, Kang-Ho;Park, Chan-Won;Ha, Sang-Keun;Kim, Geong-Gyu
    • Korean Journal of Soil Science and Fertilizer
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    • v.39 no.5
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    • pp.259-267
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    • 2006
  • Hydraulic conductivity is the rate of water flux on hydraulic gradient. The van Genuchten Mualem (VGM) model is frequently used for describing unsaturated state of soils, that is composed with the function of soil water potential and soil water content and requests various parameters. This study is to get the value of VGM parameters used Rosetta computer program based on neural network analysis method and to calculate VGM parameters. VGM parameters included Ko(effective saturated hydraulic conductivity), ${\theta}r$(residual soil water content), ${\theta}s$(saturated soil water content), L, n and m. The unsaturated hydraulic conductivity at 10 kPa was calculated by using Rosetta program. Unsaturated hydraulic conductivities of 17 soil series at 1, 3, 5, 7 kPa were also obtained by applying saturated hydraulic conductivity by disk tension infiltrometer based on Gardner and Wooding's equation. Water flow at the water potential of 3 kPa was very low except Namgye, Hagog, Baegsan, Sangju, Seogcheon, Yesan soil series. Unsaturated hydraulic conductivity at 1 kPa showed the highest value for Samgag soil series and was in order of Yesan, Hwabong, Hagog and Baegsan soil series. Those of Gacheon, Seocheon and Ugog soil series were very low. When the value by VGM was compared with the value by disc tension infiltrometer, there was a tendency with exponential function to soils without gravel but there was no tendency to soils including gravel. Conclusively, it would be limited that VGM model for unsaturated hydraulic conductivity analysis applies to Korean agricultural land including gravel and having steep slope, shallow soil depth.

LC-MS/MS analysis and anti-inflammatory effects of crude extract from Coptidis Rhizoma (황련 추출물의 LC-MS/MS 분석 및 항염증 효과)

  • Min-Jung, Kim;Ye-Jin, Yang;Kwang-Youn, Kim;Hun Hwan, Kim;Jae Dong, Son;Ju-Hye, Yang;Dong bin, Lee;Woo Hyun, Kim;Hu-Jang, Lee;Seon Been, Bak;Kwang-Il, Park
    • Herbal Formula Science
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    • v.31 no.1
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    • pp.1-10
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    • 2023
  • Objectives : The main aim of this study was to examine the LC-MS/MS used to identify phenolic compounds of CRE(Coptidis Rhizoma 70% EtOH Extract). Also, we investigated antioxidative activities and Anti-inflammatory activities. Methods : LC-MS/MS Analysis HPLC and LC-MS/MS were performed on a 1260 series HPLC system (Agilent Technologies, Inc., California, USA) and 3200 QTrap tandem mass system (Sciex LLC) operated in positive ion mode (spray voltage set at -4.5 kV). The solvent used was DW and Acetonitrile containing 0.1% formic acid, a gradient system was used at a flow rate of 0.5 mL/min for analysis, and a Prontosil C18 column (length, 250 mm; inner diameter, 4.6 mm; particle size, 5 ㎛; Phenomenex Co., Ltd., California, USA, Biochoff Chromatography) was used. The solvent conditions used in the mobile phases were 0-10 min at 10-15% B, 10-20 min at 20% B, 20-30 min at 25%, 30-40 min at 40%, 40-50 min at 70%, 50-60 min at 95%, and 60-70 min at 95%. The analysis was performed at a wavelength of 284 nm and a temperature of 35℃. The cell viability was measured using a 3-(4,5-dimethyethiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. We examined the effects of CRE on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) in a RAW 264.7 cells Results : The chemical analysis CRE by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed that Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid as phenolic components. DPPH radical scavenging activity was the inhibitory activity of CRE showed at 200 ㎍/mL a statistically significant level. MTT assay demonstrated that the CRE did not have a cytotoxic effect in RAW 264.7 and LPS-induced RAW264.7 cells. Also, CRE reduced NO production in RAW 264.7 cells stimulated with LPS. Conclusions : Based on these findings, The chemical analysis 4 major components CRE such as Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid. Moreover, we confirmed that CRE has effects antioxidant and anti-inflammatory. The results demonstrate that CRE can be used as an antioxidant and a powerful chemopreventive ingredient against inflammatory diseases.

Field Studios of In-situ Aerobic Cometabolism of Chlorinated Aliphatic Hydrocarbons

  • Semprini, Lewts
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.3-4
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    • 2004
  • Results will be presented from two field studies that evaluated the in-situ treatment of chlorinated aliphatic hydrocarbons (CAHs) using aerobic cometabolism. In the first study, a cometabolic air sparging (CAS) demonstration was conducted at McClellan Air Force Base (AFB), California, to treat chlorinated aliphatic hydrocarbons (CAHs) in groundwater using propane as the cometabolic substrate. A propane-biostimulated zone was sparged with a propane/air mixture and a control zone was sparged with air alone. Propane-utilizers were effectively stimulated in the saturated zone with repeated intermediate sparging of propane and air. Propane delivery, however, was not uniform, with propane mainly observed in down-gradient observation wells. Trichloroethene (TCE), cis-1, 2-dichloroethene (c-DCE), and dissolved oxygen (DO) concentration levels decreased in proportion with propane usage, with c-DCE decreasing more rapidly than TCE. The more rapid removal of c-DCE indicated biotransformation and not just physical removal by stripping. Propane utilization rates and rates of CAH removal slowed after three to four months of repeated propane additions, which coincided with tile depletion of nitrogen (as nitrate). Ammonia was then added to the propane/air mixture as a nitrogen source. After a six-month period between propane additions, rapid propane-utilization was observed. Nitrate was present due to groundwater flow into the treatment zone and/or by the oxidation of tile previously injected ammonia. In the propane-stimulated zone, c-DCE concentrations decreased below tile detection limit (1 $\mu$g/L), and TCE concentrations ranged from less than 5 $\mu$g/L to 30 $\mu$g/L, representing removals of 90 to 97%. In the air sparged control zone, TCE was removed at only two monitoring locations nearest the sparge-well, to concentrations of 15 $\mu$g/L and 60 $\mu$g/L. The responses indicate that stripping as well as biological treatment were responsible for the removal of contaminants in the biostimulated zone, with biostimulation enhancing removals to lower contaminant levels. As part of that study bacterial population shifts that occurred in the groundwater during CAS and air sparging control were evaluated by length heterogeneity polymerase chain reaction (LH-PCR) fragment analysis. The results showed that an organism(5) that had a fragment size of 385 base pairs (385 bp) was positively correlated with propane removal rates. The 385 bp fragment consisted of up to 83% of the total fragments in the analysis when propane removal rates peaked. A 16S rRNA clone library made from the bacteria sampled in propane sparged groundwater included clones of a TM7 division bacterium that had a 385bp LH-PCR fragment; no other bacterial species with this fragment size were detected. Both propane removal rates and the 385bp LH-PCR fragment decreased as nitrate levels in the groundwater decreased. In the second study the potential for bioaugmentation of a butane culture was evaluated in a series of field tests conducted at the Moffett Field Air Station in California. A butane-utilizing mixed culture that was effective in transforming 1, 1-dichloroethene (1, 1-DCE), 1, 1, 1-trichloroethane (1, 1, 1-TCA), and 1, 1-dichloroethane (1, 1-DCA) was added to the saturated zone at the test site. This mixture of contaminants was evaluated since they are often present as together as the result of 1, 1, 1-TCA contamination and the abiotic and biotic transformation of 1, 1, 1-TCA to 1, 1-DCE and 1, 1-DCA. Model simulations were performed prior to the initiation of the field study. The simulations were performed with a transport code that included processes for in-situ cometabolism, including microbial growth and decay, substrate and oxygen utilization, and the cometabolism of dual contaminants (1, 1-DCE and 1, 1, 1-TCA). Based on the results of detailed kinetic studies with the culture, cometabolic transformation kinetics were incorporated that butane mixed-inhibition on 1, 1-DCE and 1, 1, 1-TCA transformation, and competitive inhibition of 1, 1-DCE and 1, 1, 1-TCA on butane utilization. A transformation capacity term was also included in the model formation that results in cell loss due to contaminant transformation. Parameters for the model simulations were determined independently in kinetic studies with the butane-utilizing culture and through batch microcosm tests with groundwater and aquifer solids from the field test zone with the butane-utilizing culture added. In microcosm tests, the model simulated well the repetitive utilization of butane and cometabolism of 1.1, 1-TCA and 1, 1-DCE, as well as the transformation of 1, 1-DCE as it was repeatedly transformed at increased aqueous concentrations. Model simulations were then performed under the transport conditions of the field test to explore the effects of the bioaugmentation dose and the response of the system to tile biostimulation with alternating pulses of dissolved butane and oxygen in the presence of 1, 1-DCE (50 $\mu$g/L) and 1, 1, 1-TCA (250 $\mu$g/L). A uniform aquifer bioaugmentation dose of 0.5 mg/L of cells resulted in complete utilization of the butane 2-meters downgradient of the injection well within 200-hrs of bioaugmentation and butane addition. 1, 1-DCE was much more rapidly transformed than 1, 1, 1-TCA, and efficient 1, 1, 1-TCA removal occurred only after 1, 1-DCE and butane were decreased in concentration. The simulations demonstrated the strong inhibition of both 1, 1-DCE and butane on 1, 1, 1-TCA transformation, and the more rapid 1, 1-DCE transformation kinetics. Results of tile field demonstration indicated that bioaugmentation was successfully implemented; however it was difficult to maintain effective treatment for long periods of time (50 days or more). The demonstration showed that the bioaugmented experimental leg effectively transformed 1, 1-DCE and 1, 1-DCA, and was somewhat effective in transforming 1, 1, 1-TCA. The indigenous experimental leg treated in the same way as the bioaugmented leg was much less effective in treating the contaminant mixture. The best operating performance was achieved in the bioaugmented leg with about over 90%, 80%, 60 % removal for 1, 1-DCE, 1, 1-DCA, and 1, 1, 1-TCA, respectively. Molecular methods were used to track and enumerate the bioaugmented culture in the test zone. Real Time PCR analysis was used to on enumerate the bioaugmented culture. The results show higher numbers of the bioaugmented microorganisms were present in the treatment zone groundwater when the contaminants were being effective transformed. A decrease in these numbers was associated with a reduction in treatment performance. The results of the field tests indicated that although bioaugmentation can be successfully implemented, competition for the growth substrate (butane) by the indigenous microorganisms likely lead to the decrease in long-term performance.

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