This work was funded by the European Union Seventh Framework Program FP7/2007-2013 under grant agreement no. 265721. The authors thank the RIKILT Institute for Food Safety (NL) for their support in this project.

참고 : 다음 프로토콜은 SPE가 50 % v / V를 메탄올로 C18 흡착제 및 용출 100 ml의 물 샘플에서 수행에 대해 설명합니다. 농축 샘플 (ELISA) 키트를 분석 면역 연결 효소 분석하기 전에 10 % v / V를 메탄올에 도달하기 위해 희석한다. 1. 시약 준비 <ol><li> 물 샘플을 준비 <ol><li> 이전의 임의의 다른 단계에, 0.2 ㎛의 공극 크기의 필터를 각각 100 ml의 물 샘플을 필터링. </li><li> 물의 부피 기준으로 용액의 적절한 용적을 희석하여 목적하는 농도를 갖는 샘플 스파이크. 예를 들어, 300 μg의 / L의 농도에서 E2 참조 용액 3.3 μL 희석 E2 100 겨 / L의 물 샘플을 100㎖를 준비한다. 샘플 10 NG / L (E2)와 아군 얻기 위해이 솔루션을 10 번을 희석. 1 NG / L E2와 100 ㎖의 샘플을 얻기 위해 후자의 또 다른 열 배 희석. </li><li> G와 유리 병 (수정되지 않은 또는 아군) 필터 샘플을 배치L45 스레드. 같은 날 샘플을 사용합니다. 작은 부분을 가지고하는 초기 농도를 추정하기 위해 ELISA로 분석된다. </li></ol></li><li> 타이트한 튜브에 50 % v / V를 탈 이온화 (디) 물에 메탄올을 희석하여 리제 300 μl를 준비합니다. </li></ol> SPE 열 2. 준비 <ol><li> 첫번째 메탄올 1,600 μL 후 역상 흡착제의 40 mg의 디 물 400 μl를 첨가하여 옥타 데실 실리카 흡착제 입자의 20 ㎎ / ㎖ 현탁액을 제조. 단단히 뚜껑을 닫고 소용돌이로 교반. </li><li> 열에서 바닥 막 설치 <ol><li> 기공 크기 11 μm의 한 나일론 막을 선택 및 안티 먼지 조직의 더블 레이어에 놓습니다. 3mm 직경의 펀치로, 막에서 두 개의 작은 부분을 절단. </li><li> 평면 엔드 필터 집게와 작은 막 중 하나를 잡고 컬럼의 한쪽에 놓습니다. </li><li> 튜브 평면 하단 커넥터 나사 조이. 멤브레인은 N이고아우 제자리에 안전하게 흡수제 첨가 할 수있다. 막 배치 된 곳 끝으로 열 가리키는 몸에 화살표를 그립니다. </li></ol></li><li> 포장 흡수제 열 준비 <ol><li> 바닥을 향해 가리키는 화살표와 홀더에 열을 고정합니다. 열은 수직으로 가능한 설정해야합니다. </li><li> 루어 락 커넥터를 사용함으로써, 컬럼 튜브의 단부에 빈 10 ㎖를 일회용 주사기를 첨부. </li><li> , 20 ~ 200 μL 팁과 마이크로 피펫을 준비 100 μL에 볼륨을 설정합니다. 이 피펫 팁 포맷이 제조 될 흡착제 컬럼의 크기로 구성된다. 절차는 1 ml의 피펫에 사용되는 것과 같은 큰 피펫 팁을 더 어렵다. </li><li> 소용돌이와 흡착제 서스펜션을 선동하고, 빠르게 열에서 중앙에 100 μl를 피펫. 주입하는 동안, 모든 흡 부드럽게 피펫 팅 용액을 다른 손으로 주사기를 사용하여 멤브레인을 물마루. 본 보안 목표 명세서에서연령, 주사기를 채우는 용액 입자의 명확해야하고, 입자 베드 결국 열에서 관찰 될 수있다. </li><li> 용액에서 입자의 균일 한 현탁액을 보장하기 위해 모든 피펫 팅 단계 사이의 주식 현탁액을 교반하여이 과정을 2 번 이상 반복합니다. 그 결과 열은 흡착제의 6 밀리그램이 포함되어 있습니다. </li><li> 현탁액 300 ㎕를 로딩하고 주사기로 흡입하여 건조되었을 때, 위치에 주사기를 유지하고 다른 손의 집게를 이용하여 상부에 제 나일론 막을 배치했다. </li><li> 튜브와 제 2 커넥터를 나사와 주사기를 폐기하십시오. SPE 열을 사용할 준비가되었습니다. </li></ol></li></ol> 3. 시스템 준비 <ol><li> 루어 잠금 커넥터를 사용하여 장치의 SPE 열을 조입니다. 화살표는 장치에 표시된 것과 같은 방향을 가리키고 있어야한다. </li><li> 나사 결합 장치로 샘플을 함유하는 병을 연결그것에 GL45 안전 캡을 제공했다. </li><li> &#39;리제&#39;저수지에서 리제 200 μl를로드합니다. </li><li> 로드 &#39;희석&#39;저수지에서 디 물 800 μL. </li><li> 추출 단계에서 처리 된 물을 수집하는 폐기물 출구에 병을 놓습니다. 형식은 중요하지 않지만 볼륨 샘플 량에 대해서는 충분히 크게 할 필요가있다. </li><li> 1.5 ml의 최소 볼륨 용량 센서 출구에 작은 유리 병을 놓습니다. 풍부한 출제 및 희석 버퍼를 수집 할 때 거품이 출구에 형성되므로 1 ml의 최대 볼륨이 너무 작은 것입니다. </li><li> 압력 조정기가 수동으로 설정하여 폐쇄 위치에 확인한 역 시계 방향으로 더 이동이 불가능해질 때까지. </li></ol> 프로토 타입 4. SPE <ol><li> 뒷면의 버튼을 눌러 프로토 타입 전환합니다. </li><li> 사용자 인터페이스를 준비하고 프로그램을 선택 <ol><li> 사용자 인터페이스를 엽니 다. 고른컴퓨터의 통신 포트는 장치 목록에서 접속되고, 버튼 &#39;다음&#39;을 클릭하도록한다. </li><li> 값 PSET 580 및 dpset 30을 입력합니다. 펌프는 실행시 580 ± 30 mbar에서 가압 시스템 및 저수지의 압력을 유지하기 위해 조정됩니다. </li><li> 자동 모드를 선택합니다. </li><li> 자동 모드 구석에, 상자 &#39;구성 파일 경로&#39;의 프로그램 파일을로드. </li></ol></li><li> 압력 조정기를 조정 <ol><li> 펌프를 시작합니다. </li><li> 레그에 대한 읽기 값이 열등하지만 320 밀리바에 가까운 때까지 수동 압력 조절기를 돌립니다. </li><li> 펌프를 중지합니다. </li></ol></li><li> SPE의 절차를 시작하기 <ol><li> 자동 모드 모서리를 눌러 &#39;시작&#39;. 펌프에 전환하고 추출, 용출 및 희석 단계는 자동으로 서로를 따를 것이다. 100 ml의 시료에 대한 전체 공정 50 분에서 수행된다. </li><li>레그의 값을 확인합니다. 최적의 유량을 보장하기 위해 추출 공정 동안 350 밀리바 - 그것은 320의 범위에 있어야한다. </li><li> 분석 할 때까지 어둠 속에서 4-5 ° C에있는 작은 유리 병 및 저장을 닫습니다. 분석 물질의 열화를 방지하기 위해 다음 30 시간에서 분석을 수행한다. </li><li> 처리 된 물을 폐기하십시오. </li></ol></li><li> 시스템 청소 주의 : 시스템이 필요한 각 추출 과정은 교차 오염을 방지하기 위해 세정되어야 할 후에. <ol><li> GL (45) 유리 병에 메탄올 70 %의 v / V의 10 ml의 솔루션을 준비합니다. </li><li> SPE 열을 분리 및 커넥터와 튜브를 연결합니다. </li><li> 자동 모드에서, &#39;청소&#39;파일을 선택하고 동일한 압력 설정으로 시작합니다. </li></ol></li></ol> ELISA와 에스트라 디올 농도의 5 검출 <ol><li> 사용되는 ELISA 키트와 함께 제공되는 프로토콜에 표시된 농도 교정 샘플을 준비합니다. 하나 CALIBRA 준비메탄올 10 %의 v / v를 설정 한 것과 같은 샘플로 매트릭스, 한 교정 설정 기. </li><li> 제조업체의 지시에 따라, 플레이트에 웰에서 샘플의 필요한 양을 분배. 교정 샘플을 여과하지만 메탄올 10 % 절에서 샘플을 SPE로 처리하고, 농축되지 않은 수정되지 않은 및 아군 물 샘플을 사용하여 / V. 분석과 관련된 오류를 줄이기 위해 샘플 당 3 우물을 사용합니다. </li><li> 효소와 반응물의 첨가 용 키트, 배양 시간, 세정 및 상기 반응 구비 프로토콜의 표시를 따르. </li><li> 플레이트 리더 악기와 장비 제조업체의 지침에 따라 각 웰에 광 신호를 읽고 데이터를 수집합니다. </li><li> 메탄올 10 % v를 농축 시료를 검량선과 맞 같은 매트릭스와 초기 시료 E2의 농도를 측정하고,하기에서 플레이트 판독기 기기의 소프트웨어를 사용 / V. </li></ol>

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The authors have nothing to disclose.

면역 분석을 사용하여 분석 하였다 물 샘플의 제조를위한 새로운 방법이 제안되었다. 악기가 자동 사용자 친화적 방식으로 고체상 추출을 수행 할 수있다. 종래의 시스템에 주입 물 샘플의 여과는 매우 중요하다. 용액에 존재하는 임의의 미립자는 여전히 잠재적으로 유체 망의 막힘을 일으키고 SPE 칼럼을 방해한다. 다른 중요한 단계는 SPE 칼럼의 제조이다. 열에있는 입자의 ...

샘플은 분석 준비 과정에서 중요한 단계이다. 특히, 매트릭스 효과, 간섭의 감소, 및 분석 물의 농축 제거하여 정확한 결과를 획득하고 검출 하한에 도달하는 데 필요한. 내분비 방해 화합물 (EDCs)은 인해 생물에 대한 자신의 행동에 특별한 관심의 경우에도 환경에서 현재 매우 낮은 수준. 천연 호르몬 17β-에스트라 디올은 유럽 물 기본 지침에 의해 규제되는 우선 순위 물질 목록에 추가 할 수있는 유럽 연합 (EU)의 수질 오염 감시 목록에 현재와 경향이있다. 고체 상 추출 (SPE)는 일반적으로 화학 양 1-5 (크로마토 그래피, 질량 분광 분석) 6-9 및 면역 학적 검출 방법으로, 작은 물 오염 물질의 분석에 적용된다. 후자는 면역이 형식의 대형 다양한에서 사용할 수있는 TA에 고유 한, 환경 모니터링 분야에 대한 관심을 얻었다피검 rget 및 검출 하한에 도달한다. 6, 7, 10, 11, 각종 효소 결합 면역 분석법 (ELISA)은 시판되고 멀티 웰 플레이트에 한 번에 여러 개의 시료를 분석 가능하게한다. 절차는 몇 시간이 걸릴 수있는 연속 반응 단계에있다. 반응의 최종 생성물은 검량선에 기초하여 상기 표적 분자의 농도를 결정하기 위해 광학적으로 검출 할 수있다. 고전 SPE 절차는 용출액의 증발에 의해 흡착 프리 컨디셔닝 (pre-conditioning), 샘플 추출, 세척, 용출 및 농도를 포함한다. 이 추출물을 희석에 사용되는 용제는 검출 방법에 따라 선택된다. 면역 학적 방법에 대해서는, 강하게 유기 용매의 영향에있어서의 감도의 양. 12 회수 및 미리 농도 성능 외에도, 방법은 간단하고 효율적인 비용이 필요하다. procedur의 자동화전자는 인간의 관련 오류를 줄일 수 있습니다. 우리의 이전 연구 13에서 우리는 자동화 된 SPE에 대한 우리의 프로토 타입을 소개하고, 우리의 방법은 해수 시료에서 자연 호르몬 17β-에스트라 디올의 분석에 적용 하였다. 본 동영상으로 우리는 SPE 및 면역 반응에 의해 감지와의 특별한 호환성 기존의 오프라인과 온라인에 비해 우리의 방법의 기술적 인 장점을 강조하고 싶습니다. 우리는 17β-에스트라 디올의 검출 물 샘플에 적용된 프로토콜을 설명한다. SPE는 옥타 데실 실리카 (C18) 흡수제 및 용출 단계가 희석 된 메탄올 행한다 행한다.

<table><tbody><tr><td>Filter membrane 0.2 &amp;mu;m pore size</td><td>Merck Millipore</td><td>GNWP04700</td><td>For sample filtration</td></tr><tr><td>Nylon membrane 11 &amp;mu;m pore size</td><td>Merck Millipore</td><td>NY1104700</td><td>For SPE column</td></tr><tr><td>Disposable biopsy punch 5 mm</td><td>Medical Budget</td><td>39302439</td><td></td></tr><tr><td>Nucleodur C18 ec&amp;nbsp;</td><td>Macherey Nagel</td><td>713550.01</td><td>50 &amp;mu;m particle diameter</td></tr><tr><td>Synthetic sea water</td><td>Sigma Aldrich</td><td>SSWS500-500ML</td><td></td></tr><tr><td>Methanol</td><td>VWR</td><td></td><td></td></tr><tr><td>17beta-estradiol standard</td><td>Enzo Life Science</td><td></td><td>300 ng/ml</td></tr><tr><td>17beta-estradiol ELISA kit</td><td>Enzo Life Science</td><td>ADI-900-008</td><td>96 wells, range 30 - 3,000 ng/L</td></tr></tbody></table>

a simple method for automated solid phase extraction of water samples for immunological analysis of small pollutants

환경 물 샘플의 고체 상 추출 (SPE)을위한 새로운 방법을 제안한다. 개발 된 프로토 타입은 비용 효율적이고 사용자 친화적이며, 신속한 자동화 및 간단한 SPE을 수행 할 수 있습니다. 예비 농축액 낮은 유기 용매 함량, 면역 측정법에 의한 분석과 호환 가능하다. 방법은 100 mL의 물을 샘플에 천연 호르몬 17β-에스트라 디올의 추출 및 미리 농도를 설명한다. 역상 SPE는 옥타 - 실리카 흡착제로 수행되고 용출 메탄올 50 % V / V를 200 μL로 이루어집니다. 용출액은 메탄올의 양을 낮추는 디 물을 첨가하여 희석한다. 수동 SPE 칼럼을 준비한 후, 전체 절차는 1 시간 이내에 자동으로 수행된다. 프로세스의 끝에서, 에스트라 디올 농도 상업용 효소 결합 면역 흡착제 분석법 (ELISA)을 이용하여 측정된다. 100 배 사전 농도를 달성하고 10 %의 V / V에서 메탄올 내용. 분자의 전체 복구1 NG로 달성된다 / L은 드 이온화 및 합성 해수 샘플을 아군.

흡착재 충전부의 재현성이 건조에 의해 평가하고 유리 바이알에 피펫 흡착제 가중치 및 초기 및 사전에도 2. 농도로 나타낸 바와 같이 결과는, 100 mL의 시료에 대해 시험 하였다 주입시의도 1. 재현성에 도시 된 농축 된 샘플은 17β-에스트라위한 상용 ELISA 키트를 사용하여 측정하고,도 3에 도시되어 아군. <p class="jove_content" fo:keep-toget...

Watch this Scientific Journal Video about A Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants at JoVE.com

A Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants

A protocol for the extraction and pre-concentration of estradiol from water samples by using an automated and miniaturized system is presented.

작은 오염 물질의 면역 학적 분석을위한 물 샘플의 자동화 된 고체상 추출을위한 간단한 방법

A new method for solid phase extraction (SPE) of environmental water samples is proposed. The developed prototype is cost-efficient and user friendly, and ...

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13.6K 조회수.  Centre Suisse d'Electronique et de Microtechnique. A protocol for the extraction and pre-concentration of estradiol from water samples by using an automated and miniaturized system is presented.

비디오: A Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants

Automated Gel Size Selection to Improve the Quality of Next-generation Sequencing Libraries Prepared from Environmental Water Samples

Next-generation sequencing of environmental samples can be challenging because of the variable DNA quantity and quality in these samples. High quality DNA libraries are needed for optimal results from next-generation sequencing. Environmental samples such as water may have low quality and quantities of DNA as well as contaminants that co-precipitate with DNA. The mechanical and enzymatic processes involved in extraction and library preparation may further damage the DNA. Gel size selection enables purification and recovery of DNA fragments of a defined size for sequencing applications. Nevertheless, this task is one of the most time-consuming steps in the DNA library preparation workflow. The protocol described here enables complete automation of agarose gel loading, electrophoretic analysis, and recovery of targeted DNA fragments. 
In this study, we describe a high-throughput approach to prepare high quality DNA libraries from freshwater samples that can be applied also to other environmental samples. We used an indirect approach to concentrate bacterial cells from environmental freshwater samples; DNA was extracted using a commercially available DNA extraction kit, and DNA libraries were prepared using a commercial transposon-based protocol. DNA fragments of 500 to 800 bp were gel size selected using Ranger Technology, an automated electrophoresis workstation. Sequencing of the size-selected DNA libraries demonstrated significant improvements to read length and quality of the sequencing reads.

This manuscript describes an automated gel size selection approach for purifying DNA fragments for next-generation sequencing. The Ranger Technology provides complete automation of the entire process of agarose gel loading, electrophoretic analysis, and recovery of targeted DNA fragments allowing for high-throughput and high quality next-generation sequencing libraries.

자동 젤 사이즈 선택은 환경 물 샘플에서 준비 차세대 시퀀싱 라이브러리의 질을 향상시키기 위해

Next-generation sequencing of environmental samples can be challenging because of the variable DNA quantity and quality in these samples. High quality DNA ...

연구

환경과학

Preparation and Testing of Impedance-based Fluidic Biochips with RTgill-W1 Cells for Rapid Evaluation of Drinking Water Samples for Toxicity

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity sensor. A monolayer of RTgill-W1 cells forms on the sensing electrodes enclosed within the biochips. The biochips are then used for testing in a field portable electric cell-substrate impedance sensing (ECIS) device designed for rapid toxicity testing of drinking water. The manuscript further describes how to run a toxicity test using the prepared biochips. A control water sample and the test water sample are mixed with pre-measured powdered media and injected into separate channels of the biochip. Impedance readings from the sensing electrodes in each of the biochip channels are measured and compared by an automated statistical software program. The screen on the ECIS instrument will indicate either "Contamination Detected" or "No Contamination Detected" within an hour of sample injection. Advantages are ease of use and rapid response to a broad spectrum of inorganic and organic chemicals at concentrations that are relevant to human health concerns, as well as the long-term stability of stored biochips in a ready state for testing. Limitations are the requirement for cold storage of the biochips and limited sensitivity to cholinesterase-inhibiting pesticides. Applications for this toxicity detector are for rapid field-portable testing of drinking water supplies by Army Preventative Medicine personnel or for use at municipal water treatment facilities.

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial cells for use in a field portable electric cell-substrate impedance sensor. The protocol for running a rapid drinking water toxicity test with the sensor is also described.

준비 및 시험 독성에 대한 신속한 평가 음주의 물 샘플에 대한 RTgill-W1 세포와 유체 바이오칩을 임피던스 기반

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity ...

Characterization and Application of Passive Samplers for Monitoring of Pesticides in Water

Five different water passive samplers were calibrated under laboratory conditions for measurement of 124 legacy and current used pesticides. This study provides a protocol for the passive sampler preparation, calibration, extraction method and instrumental analysis. Sampling rates (RS) and passive sampler-water partition coefficients (KPW) were calculated for silicone rubber, polar organic chemical integrative sampler POCIS-A, POCIS-B, SDB-RPS and C18 disk. The uptake of the selected compounds depended on their physicochemical properties, i.e., silicone rubber showed a better uptake for more hydrophobic compounds (log octanol-water partition coefficient (KOW) &#62; 5.3), whereas POCIS-A, POCIS-B and SDB-RPS disk were more suitable for hydrophilic compounds (log KOW &#60; 0.70).

A protocol about the characterization and application of five different passive sampling devices is presented.

물에 농약의 모니터링을위한 특성화 및 패시브 샘플러의 응용 프로그램

Five different water passive samplers were calibrated under laboratory conditions for measurement of 124 legacy and current used pesticides. This study ...

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and detritus, and excreting particulate and dissolved compounds, they serve as important agents for benthic-pelagic coupling. Accurately measuring the compounds removed and excreted by suspension feeders (such as sponges, ascidians, polychaetes, bivalves) is crucial for the study of their physiology, metabolism, and feeding ecology, and is fundamental to determine the ecological relevance of the nutrient fluxes mediated by these organisms. However, the assessment of the rate by which suspension feeders process particulate and dissolved compounds in nature is restricted by the limitations of the currently available methodologies. Our goal was to develop a simple, reliable, and non-intrusive method that would allow clean and controlled water sampling from a specific point, such as the excurrent aperture of benthic suspension feeders, in situ. Our method allows simultaneous sampling of inhaled and exhaled water of the studied organism by using minute tubes installed on a custom-built manipulator device and carefully positioned inside the exhalant orifice of the sampled organism. Piercing a septum on the collecting vessel with a syringe needle attached to the distal end of each tube allows the external pressure to slowly force the sampled water into the vessel through the sampling tube. The slow and controlled sampling rate allows integrating the inherent patchiness in the water while ensuring contamination free sampling. We provide recommendations for the most suitable filtering devices, collection vessel, and storing procedures for the analyses of different particulate and dissolved compounds. The VacuSIP system offers a reliable method for the quantification of undisturbed suspension feeder metabolism in natural conditions that is cheap and easy to learn and apply to assess the physiology and functional role of filter feeders in different ecosystems.

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

We introduce the VacuSIP, a simple, non-intrusive, and reliable method for clean and accurate point sampling of water. The system was developed and evaluated for the simultaneous collection of the water inhaled and exhaled by benthic suspension feeders in situ, to cleanly measure removal and excretion of particulate and dissolved compounds.

VacuSIP, 개선 INEX 방법에 대한<em&gt; 현장에서</em&gt; 미립자의 측정 및 용존 화합물은 액티브 서스펜션 피더에 의해 처리

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and ...

A Modified EPA Method 1623 that Uses Tangential Flow Hollow-fiber Ultrafiltration and Heat Dissociation Steps to Detect Waterborne Cryptosporidium and Giardia spp.

Cryptosporidium and Giardia species are two of the most prevalent protozoa that cause waterborne diarrheal disease outbreaks worldwide. To better characterize the prevalence of these pathogens, EPA Method 1623 was developed and used to monitor levels of these organisms in US drinking water supplies 12. The method has three main parts; the first is the sample concentration in which at least 10 L of raw surface water is filtered. The organisms and trapped debris are then eluted from the filter and centrifuged to further concentrate the sample. The second part of the method uses an immunomagnetic separation procedure where the concentrated water sample is applied to immunomagnetic beads that specifically bind to the Cryptosporidium oocysts and Giardia cysts allowing for specific removal of the parasites from the concentrated debris. These (oo)cysts are then detached from the magnetic beads by an acid dissociation procedure. The final part of the method is the immunofluorescence staining and enumeration where (oo)cysts are applied to a slide, stained, and enumerated by microscopy.
Method 1623 has four listed sample concentration systems to capture Cryptosporidium oocysts and Giardia cysts in water: Envirochek filters (Pall Corporation, Ann Arbor, MI), Envirochek HV filters (Pall Corporation), Filta-Max filters (IDEXX, Westbrook, MA), or Continuous Flow Centrifugation (Haemonetics, Braintree, MA). However, Cryptosporidium and Giardia (oo)cyst recoveries have varied greatly depending on the source water matrix and filters used1,14. A new tangential flow hollow-fiber ultrafiltration (HFUF) system has recently been shown to be more efficient and more robust at recovering Cryptosporidium oocysts and Giardia cysts from various water matrices; moreover, it is less expensive than other capsule filter options and can concentrate multiple pathogens simultaneously1-3,5-8,10,11. In addition, previous studies by Hill and colleagues demonstrated that the HFUF significantly improved Cryptosporidium oocysts recoveries when directly compared with the Envirochek HV filters4. Additional modifications to the current methods have also been reported to improve method performance. Replacing the acid dissociation procedure with heat dissociation was shown to be more effective at separating Cryptosporidium from the magnetic beads in some matrices9,13 .
This protocol describes a modified Method 1623 that uses the new HFUF filtration system with the heat dissociation step. The use of HFUF with this modified Method is a less expensive alternative to current EPA Method 1623 filtration options and provides more flexibility by allowing the concentration of multiple organisms.

A Modified EPA Method 1623 that Uses Tangential Flow Hollow-fiber Ultrafiltration and Heat Dissociation Steps to Detect Waterborne Cryptosporidium and Giardia spp.

This protocol describes the use of a tangential flow hollow-fiber ultrafiltration sample concentration system and a heat dissociation as alternative steps for the detection of waterborne Cryptosporidium and Giardia species using EPA Method 1623.

Waterborne를 검색하는 접선 흐름 빈 섬유 Ultrafiltration 및 열 분해 단계를 사용 수정일 EPA 방법 1,623<em&gt; Cryptosporidium</em&gt;과<em&gt; Giardia SPP.</em

Cryptosporidium and Giardia species are two of the most prevalent protozoa that cause waterborne diarrheal disease outbreaks worldwide. To better ...

면역학 및 감염병학

Identifying Per- and Polyfluorinated Chemical Species with a Combined Targeted and Non-Targeted-Screening High-Resolution Mass Spectrometry Workflow

Historical and emerging per- and polyfluoroalkyl substances (PFASs) have garnered significant interest from the public and government agencies from the local to federal levels. The continuing evolution of PFAS chemistries presents a challenge to the environmental monitoring, where ongoing development of targeted methods necessarily lags the discovery of new chemical compounds. There is a need, therefore, to have forward-looking methodologies that can detect emerging and unexpected compounds, monitor these species over time, and resolve details of their chemical structure to enable future work in human health. To this end, non-targeted analysis by high-resolution mass spectrometry offers a broad base detection approach that can be combined with almost any sample preparation scheme and provides significant capabilities for compound identification after detection. Herein, we describe a solid-phase extraction (SPE) based sample concentration method tuned for shorter chain and more hydrophilic PFAS chemistries, such as per fluorinated ether acids and sulfonates, and describe analysis of samples prepared in this fashion in both targeted and non-targeted modes. Targeted methods provide superior quantification when reference standards are available but are intrinsically limited to expected compounds when performing analysis. In contrast, a non-targeted approach can identify the presence of unexpected compounds and provide some information about their chemical structure. Information about chemical features can be used to correlate compounds across sample locations and track abundance and occurrence over time.

Here, we present a protocol for the sequential targeted quantification and non-targeted analysis of fluorinated compounds in water by mass spectrometry. This methodology provides quantitative levels of known fluorochemical compounds and identifies unknown chemicals in related samples with semi-quantitative estimates of their abundance.

당 식별-및 결합된 대상 및 비 대상 심사 고해상도 질량 분석 워크플로 Polyfluorinated 화학 종

Historical and emerging per- and polyfluoroalkyl substances (PFASs) have garnered significant interest from the public and government agencies from the ...

Fabrication of a Dipole-assisted Solid Phase Extraction Microchip for Trace Metal Analysis in Water Samples

This paper describes a fabrication protocol for a dipole-assisted solid phase extraction (SPE) microchip available for trace metal analysis in water samples. A brief overview of the evolution of chip-based SPE techniques is provided. This is followed by an introduction to specific polymeric materials and their role in SPE. To develop an innovative dipole-assisted SPE technique, a chlorine (Cl)-containing SPE functionality was implanted into a poly(methyl methacrylate) (PMMA) microchip. Herein, diverse analytical techniques including contact angle analysis, Raman spectroscopic analysis, and laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) analysis were employed to validate the utility of the implantation protocol of the C-Cl moieties on the PMMA. The analytical results of the X-ray absorption near-edge structure (XANES) analysis also demonstrated the feasibility of the Cl-containing PMMA used as an extraction medium by virtue of the dipole-ion interactions between the highly electronegative C-Cl moieties and the positively charged metal ions.

The fabrication protocol of a dipole-assisted solid phase extraction microchip for the trace metal analysis is presented.

물 샘플에서 추적 금속 분석을위한 쌍극자를 이용한 고체 단계 추출 마이크로 칩의 제작

This paper describes a fabrication protocol for a dipole-assisted solid phase extraction (SPE) microchip available for trace metal analysis in water ...

생체공학

Electrowetting-based Digital Microfluidics Platform for Automated Enzyme-linked Immunosorbent Assay

Electrowetting is the effect by which the contact angle of a droplet exposed to a surface charge is modified. Electrowetting-on-dielectric (EWOD) exploits the dielectric properties of thin insulator films to enhance the charge density and hence boost the electrowetting effect. The presence of charges results in an electrically induced spreading of the droplet which permits purposeful manipulation across a hydrophobic surface. Here, we demonstrate EWOD-based protocol for sample processing and detection of four categories of antigens, using an automated surface actuation platform, via two variations of an Enzyme-Linked Immunosorbent Assay (ELISA) methods. The ELISA is performed on magnetic beads with immobilized primary antibodies which can be selected to target a specific antigen. An antibody conjugated to HRP binds to the antigen and is mixed with H2O2/Luminol for quantification of the captured pathogens. Assay completion times of between 6 and 10 min were achieved, whilst minuscule volumes of reagents were utilized.

Electrowetting-based digital microfluidic is a technique that utilizes a voltage-driven change in the apparent contact angle of a microliter-volume droplet to facilitate its manipulation. Combining this with functionalized magnetic beads enables the integration of multiple laboratory unit operations for sample preparation and identification of pathogens using Enzyme-linked Immunosorbent Assay (ELISA).

자동화된 효소 연계 면역흡착제 분석기용 전기습식 기반 디지털 미세유체 분석 플랫폼

Electrowetting is the effect by which the contact angle of a droplet exposed to a surface charge is modified. Electrowetting-on-dielectric (EWOD) exploits ...

Solid-Liquid Extraction

Source: Laboratory of Dr. Jay Deiner &#8212; City University of New York
Extraction is a crucial step in most chemical analyses. It entails removing the analyte from its sample matrix and passing it into the phase required for spectroscopic or chromatographic identification and quantification. When the sample is a solid and the required phase for analysis is a liquid, the process is called solid-liquid extraction. A simple and broadly applicable form of solid-liquid extraction entails combining the solid with a solvent in which the analyte is soluble. Through agitation, the analyte partitions into the liquid phase, which may then be separated from the solid through filtration. The choice of solvent must be made based on the solubility of the target analyte, and on the balance of cost, safety, and environmental concerns.

Solid-Liquid Extraction: Principle, Procedure, Applications

고체 액체 추출

Source: Laboratory of Dr. Jay Deiner &#8212; City University of New York
Extraction is a crucial step in most chemical analyses. It entails removing the ...

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화학

유기 화학

Extraction of Biomarkers from Sediments - Accelerated Solvent Extraction

Source: Laboratory of Jeff Salacup&#160;- University of Massachusetts Amherst
The distribution of a group of organic biomarkers called glycerol-dialkyl glycerol-tetraethers (GDGTs), produced by a suite of archaea and bacteria, were found in modern sediments to change in a predictable manner in response to air or water temperature1,2. Therefore, the distribution of these biomarkers in a sequence of sediments of known age can be used to reconstruct the evolution of air and/or water temperature on decadal to millennial timescales (Figure 1). The production of long high-resolution records of past climates, called paleoclimatology, depends on the rapid analysis of hundreds, possibly thousands of samples. Older extraction techniques, such as sonication or Soxhlet, are too slow. However, the newer Accelerated Solvent Extraction technique was designed with efficiency in mind.
<img alt="Figure 1" src="/files/ftp_upload/10097/10097fig1.jpg" /> 
Figure 1. An example of a paleoclimate record showing changes in sea surface temperature (SST) in the eastern Mediterranean Sea during the past ~27,000 years3. This record comprises ~115 samples and is based on the isoprenoidal GDGT-based TEX86 SST proxy.

Accelerated Solvent Extraction of Total Lipid from Sediments

침전물에서 생물지표 추출-가속 용매 추출

Source: Laboratory of Jeff Salacup&#160;- University of Massachusetts Amherst
The distribution of a group of organic biomarkers called glycerol-dialkyl ...

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