Microfluidic Automation in Analytical Chemistry

Column switching, automation of SPE-based sample conditioning, and post-column derivatization are forms of fluidic automation familiar to many analytical chemists. Automation of SPE sample cleanup has been employed extensively when large numbers of samples must be processed, such as in the analysis of pesticides in foods and, more recently, has facilitated adoption of modern methodology for paralytic shellfish toxins (PST’s) in UK shellfish monitoring. Similarly, post-column derivatization has long been a mainstay in the study of the PST’s and will soon form the basis for a new official method for shellfish toxin control. These approaches will be covered in our external workshop on LC detection of PST’s.

Sequential Injection Analysis

At the Annual Meeting, we will also offer laboratory sessions on another form of fluidic automation that is a less familiar but very capable technology. Sequential injection analysis (SIA) is a very high throughput analytical tool that is well suited to miniaturization. SIA is a microfluidic approach to automating chemistries that has been employed extensively in the development of unattended monitoring systems such as in process control and environmental monitoring. It is the successor to flow injection analysis (FIA) and one of our keynote speakers, Jaromir Ruzicka, is a co-inventor of both of these approaches to automating chemistries. In our laboratory workshop we will show that SIA is also well suited to the development of assays using expensive reagents such as enzymes, and is a green approach, in that very little waste is generated. SIA systems also have a small footprint, and could easily be mounted in portable enclosures. More information is available at www.flowinjection.com
SIA will be demonstrated for the microfluidic automation of an enzymatic assay for histamine and the high speed analysis of other targets. Admission to the SIA laboratory workshop is included in the registration fee for the Annual Meeting.

Instructor Hugo Oliveira

Hugo Oliveira obtained his degree in Pharmaceutical Sciences at University of Porto (Portugal) in 2003. This included a final training period in Analytical Chemistry area as student of the “Erasmus” exchange program (2003). He worked in the development of optical sensors under supervision of Dr. Julian Alonso at the Sensors and Biosensors Group of Autonomous University of Barcelona (Spain). In the same year he began the European Master Course in Environmental Analytical Chemistry at the University of Porto in collaboration with the University of Aberdeen (United Kingdom). During the case-study module of this master, he was integrated in the Air Quality Survey project at Aberdeen Harbour managed by Dr. Iain Marr, participating in the development and application of analytical methodologies for atmospheric pollutants analysis. In January of 2005 received his Master in Environmental Analytical Chemistry after a research project dealing with automatic flow based method for the determination of phenolic compounds in water samples, carried out at the University of Porto. During 2005 he worked as a teacher in technical training courses in Analytical Chemistry and obtained a PhD grant from Portuguese Science Foundation. Since 2006 he has been working at his PhD program centered in the development of new environmentally friendly automatic methodologies for sample preparation coupled to liquid chromatography (LC), supervised by Prof. Marcela Segundo and Prof. José Luís Costa Lima. During this period he visited the University of Balearic Islands (Spain) and worked with Prof. Manuel Mirò and Prof. Victor Cerdà. As result of this collaboration, new strategies for automatic sample preconcentration and cleanup hyphenated with LC applied to environmental and foodstuff samples were recently published.

His present research interests are focused in the development of new automation strategies for renewable solid-phase extraction and chemical assays based on microfluidic systems. Hugo is currently a visitor of the Department of Oceanography of the University of Hawai’i at Mañoa, working in collaboration with Prof. Jaromir Ruzicka and Prof. Christopher Measures in the development of new microfluidic based methods for the determination of trace metals in seawater.

Molecular Methods in Food Microbiology

Microbiology goes Molecular

Schedule Here

PCR Pathratrix Training (On-site at Annual Meeting)

June 22-23, 2010

Jinxin Hu, Ph.D., FDA, Santos Camara, FDA

Marcie van Wart, Matrix MicroScience

Pathatrix is a rapid detection method for pathogens, spoilage bacterium and viruses in food, environmental and bio-threat samples. It uses re-circulating immuno-magnetic separation (RIMS) technology that relies on the use of antibody coated paramagnetic particles to selectively bind and purify the target organism from a comprehensive range of complex matrices. By re-circulating the sample over a “capture phase,” comprised of immobilized antibody coated magnetic beads, this increases the sensitivity of the capture. A high volume wash enables the efficient removal of the sample matrices, non-specific microorganisms and PCR inhibitors. Pathatrix instrument shown here and also diagrams of the steps attaching the capture phase and inserting consumable into the catridge.

In conjunction with real time PCR Pathatrix can provide results within hours. It is also used to improve the performance of other rapid methods such as PCR, lateral flow, ELISA and chromgenic media by reducing or eliminating the need for lengthy pre-enrichment and/or selective enrichment steps.

In this training course, we will discuss the approaches of using antibody coated magnetic particles as well as cationic beads to aid the capture of pathogens from food. After the lecture and discussion, participants will perform a test using Pathatrix involving 4 major steps including sample preparation, pre-incubation, placing the sample in the Pathatrix, and washing and elution. Real time PCR and culture methods will be used to identify the pathogens retrieved from Pathatrix system.

Applied Biosystems Pathogen Detection System for Real-Time PCR Detection of E. coli O157:H7

(On-site at Annual Meeting)

June 23, 2010

Instructors: Sharron Henne, Justin Lao

Applied Molecular Testing Business, Molecular Biology Division

This workshop will provide an overview of the Applied Biosystems Pathogen Detection System and its use for detecting E. coli O157:H7 in food and environmental samples.  The workshop will feature:

- An introduction to real-time PCR and its use for pathogen detection

- Manual and automated sample preparation methods for nucleic acids extraction

- A real-time PCR assay for detection of E. coli O157:H7

- Software that fully automates amplification, detection, data collection and analysis

- An onsite demo of the full workflow from sample preparation to data analysis

Click Here for Photo of Applied Biosystems instrument.

A Pragmatic Approach for Ultra Trace Mercury Analysis

Are You Getting Value from Your Analyzer Workspace and Laboratory Environment?

Jeff Forsberg, CETAC Technologies

Getting the best business value from an instrument requires knowledge of the instrument and its operating abilities, any attachments, the sample including sampling procedure and the software. All of these elements must be pulled together by a skilled and knowledgeable analyst. Unfortunately, this is not always the case in many organizations.

This workshop will not assist the analyst with knowing their specific instrument, but we will focus on setting up a laboratory for low-level analysis, which will include discussions on lab preparations, personnel skill levels and training.

There will also be discussions of EPA 1631 and 245.7 methodologies and implementation using a no-nonsense approach. Most, if not all, of the considerations that must be taken into account prior to diving into low-level mercury analysis will be discussed. This low-level approach will also assist in collecting precise data at the ppb levels. Most labs can accomplish this on a shoestring budget—they just need to be pointed in the right direction. The workshop will also include hands-on training for building purging apparatus and purging stations to assist in contaminant-free reagents.

So let’s get geared up for low-level analysis—come find out how to…

Purge Apparatus Supplies

Sample Pretreatment Automation

UPS, Tacoma – Pac NW AOAC, and Shoreline Laboratory

“Rapid Testing” is an important theme in this year’s Annual Meeting and, of course, rapid turnaround of analytical results is long sought after in the food industry and by regulators. As many of us in food safety realize, although a determinative step may be rapid enough, there are often sample handling needs that add significantly to the overall time required for analysis. When timely results are critical automation can be the answer. At this year’s meeting we will see automated sample pretreatment assume many forms, ranging from the immunomagnetic capture and purification used in pathogen detection, to automated solid phase extraction (SPE) cleanup prior to chromatographic analysis, and even the integrated components of a “Lab-On-Valve” sequential injection analyzer.

The following lectures and hands-on laboratory sessions (both onsite and in workshop following the Annual Meeting) will address automated sample pretreatment and/or fluidics automaton.

Immunomagnetic Capture:

PCR Pathatrix Training – Hands-on course on this immunomagnetic capture and PCR technology by Jinxin Hu and Santos Camara, FDA, and Marcie van Wart, Matrix Microscience. (On-site lab training as part of Microbiology goes Molecular)

Flow Injection, Sequential Injection:

From Beakers to Microfluidics – Lecture by Jaromir Ruzicka, University of Hawaii, USA (Keynote Talk)

Lab-On-Valve Technology and Sequential Injection Analysis – Hands-on course in our onsite laboratory training, including introductory lecture by Hugo Oliveira, University of Hawaii, USA (Onsite lab training, Microfluidic Automation in Analytical Chemistry) and introduced in Dr. Ruzicka’s lecture.

Automation of Solid Phase Extraction Sample Pretreatment:

Hands-on Lab Sessions on SPE Sample Pretreatment Automation – (Paralytic Shellfish Toxins by HPLC) Laboratory Workshop – June 23 and 24, at the Washington State Public Health Laboratory (Separate registration required).

High Throughput Quantitation of Saxitoxin in Human Urine Using 96-well Solid Phase Extraction. Rudy Johnson, CDC, Georgia, USA. (Marine Toxins Seminar)

Automation of Sample Preparation. -John de Kanel, Caliper LS, USA (Marine Toxins Seminar)

Automation Solutions for the Extraction and Clean-up of Residues from Food and Water – Lecture by Michael Halvorson, Gilson, Inc. (Pesticides and Industrial Contaminants Seminar)

Semi Automated, Pre-Column Oxidation Method 2005.06 for PSTs – Monitoring of UK Shellfish – Lecture by Robert Hatfield, Cefas, UK (Marine Toxins Seminar)

Marine and Freshwater Toxins – Validations, Implementations, Fluidics Automation, and the Oxidations behind the LC Fluorescence Methods for the Paralytic Shellfish Toxins
Lecture by Jim Hungerford, FDA/ORA, USA (Marine Toxins Seminar)

Principles of Pre-column Oxidation Method 2005.06 for the Paralytic Shellfish Toxins, Refinements Made at FDA, and Course Overview- (Overview lecture on 2005.06) – Lecture by Stacey (Etheridge)
DeGrasse, FDA/CFSAN (Marine Toxins Seminar)