Grantee Research Project Results
2004 Progress Report: FRIENDS Analytical Toxicology Core Facility
EPA Grant Number: R829390C003Subproject: this is subproject number 003 , established and managed by the Center Director under grant R829390
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
Center: Water Innovation Network for Sustainable Small Systems
Center Director: Reckhow, David A.
Title: FRIENDS Analytical Toxicology Core Facility
Investigators: Kostyniak, Paul J. , Olson, James
Current Investigators: Kostyniak, Paul J.
Institution: The State University of New York at Buffalo
EPA Project Officer: Aja, Hayley
Project Period: October 17, 2001 through October 16, 2002
Project Period Covered by this Report: October 17, 2003 through October 16, 2004
RFA: Centers for Children's Environmental Health and Disease Prevention Research (2001) RFA Text | Recipients Lists
Research Category: Human Health , Children's Health
Objective:
The objective of the Fox River Environment and Diet Study (FRIENDS) Analytical Toxicology Core Facility is to provide analytical support to both the epidemiological (R829390C002) and laboratory research projects (R829390C001 and R829390C004) of the FRIENDS Children’s Environmental Health Center. This core facility support involves the development of new and improved analytical techniques and analysis of human and animal study samples. All procedures are controlled by a strict QA/QC program. The core facility has extensive experience in analyzing specific congeners of polychlorinated biphenyls (PCBs), pesticides, and heavy metals at levels normally encountered in biological and experimental samples. The core facility also provides analysis of dioxin-like activity in biological samples using a reporter gene bioassay for quantitation of total dioxin (i.e., TCDD) toxic equivalents.
Progress Summary:
Advancements in Analytical Methods
Methods for analysis of specific PCB congeners and pesticides are being improved constantly to enhance sensitivity and efficiency of sample analysis. The following method improvements were accomplished during the current grant year:
Accelerated Solvent Extraction (ASE). A Dionex Accelerated Solvent Extraction system was purchased for the automated extraction and cleanup of serum samples, fish tissue samples, and rat tissues (brain, liver). Methods have been developed that have resulted in much less sample preparation time. In addition, sample extracts are much cleaner (lower PCB background) because less glassware is being used and the ability to preclean the adsorbents/filters used to extract samples is improved.
Solid Phase Extraction. We evaluated the use of disposable solid phase extraction (SPE) columns (500 mg) for the cleanup of sample extracts. Previous methods required use of chromatographic glass columns and larger volumes (10 g) of Florisil. Cartridges from different manufacturers where evaluated for phthalate/contamination and elution patterns were determined for PCBs. The SPE cartridges also were incorporated into the method used for the cleanup of rat tissue samples.
New Laboratory Glassware Washer. A Miele laboratory glassware washer was installed, which greatly reduces the amount of time spent on cleaning glassware and extraction cells. Previously, all glassware was cleaned by hand. There now is less variability in the cleanliness of glassware (i.e., PCB/phthalate ester background levels), which is a common problem for environmental laboratory analysis.
ChemStation Software for Data Acquisition and Processing. ChemStation software was installed on a new computer system, which will allow for complete control of instrumental parameters for the HP6890 Gas Chromatograph, data acquisition, calibration, and processing of chromatographic data. In addition, we are evaluating the use of Retention Time Locking software, which allows for automated adjustments in the column head pressure to maintain constant retention times. This is critical in the analysis of PCB congeners, where retention times are used for identification and any shifts require recalibration. It is common for retention times to shift over time and with routine maintenance of the system.
Zymark TurboVap II - Concentration Workstation. The Zymark system was purchased and provides an automated alternative to nitrogen blow-down evaporation for sample concentration prior to gas chromatography analysis. This instrument allows for the unattended concentration of samples, improves throughput, frees up resources, and improves quality control.
Automated Lipid Analysis. Methods for the determination of total serum lipids were established using automated standard enzymatic procedures. Total serum lipids are calculated by summation of the individual lipid components from the measurement of total cholesterol, free cholesterol, triglycerides, and phospholipids.
Analysis of Center Samples
A total of 1,500 analyses were performed during Year 3 of the project for specific PCB congeners and pesticides. This includes actual samples (human serum, rat brain tissues, and fish tissues) received from Center projects, as well as samples for the development of new methods, evaluation of new sample matrices and new equipment, and quality control/calibration samples. The number of study samples analyzed for PCB congeners and pesticides was 56 human serum samples, 6 rat brain samples, and 20 fish samples. Additionally, the 56 human serum samples were analyzed for methylmercury. One additional human serum sample, and 31 rat brain, 31 rat liver, and 24 rat serum samples will be analyzed for PCB congeners and pesticides by the end of the current grant year.
Future Activities:
Sample Analysis
Analyze samples in support of Center projects including human serum and rat brain, liver, and serum samples for specific PCB congeners, DDE, Mirex, and methylmercury.
Method Improvements
The primary focus of method development will be for assessing the concentration of polybrominated diphenyl ethers (PBDEs) in human serum samples. We will evaluate current methods to determine extraction efficiency, chromatographic cleanup elution profiles, and gas chromatographic conditions for PDBEs. Methods then will be validated for specific sample matrices.
Evaluation of a new High Volume Injection System for GC Analysis of PCB Congeners. A high volume injection system will be installed, which will allow for a higher sample volume of extracts to be injected into the gas chromatograph. The objective is to increase the size of the sample extract being analyzed, resulting in lower detection limits for samples. This is particularly important for rat brain tissue samples, where sample weights are only in the mg range. This also may reduce sample preparation time by decreasing the amount of solvent evaporation necessary for final sample extracts.
Development of Methodologies for the Determination of Hydroxylated PCBs. Methods for the extraction and determination of the major hydroxylated PCB metabolites will be developed as time permits.
Supplemental Keywords:
children’s health, disease and cumulative effects, ecological risk assessment, susceptibility, sensitive population, toxicology, Fox River, PCBs, exposure assessment, heavy metals, methylmercury, neurotoxicity, pesticides, fish consumption,, RFA, Scientific Discipline, Health, ENVIRONMENTAL MANAGEMENT, Geographic Area, POLLUTANTS/TOXICS, Midwest, Toxicology, Health Risk Assessment, Epidemiology, Chemistry, Chemicals, Risk Assessments, Susceptibility/Sensitive Population/Genetic Susceptibility, Disease & Cumulative Effects, Ecological Risk Assessment, Molecular Biology/Genetics, Children's Health, genetic susceptability, Biology, Risk Assessment, human data, neurotoxic, behavioral assessment, PCBs, pesticides, animal model, electrochemical detection, children, neurotoxicity, motor development, behavioral deficits, methylmercury, PCB, cognitive development, human exposure, Wisconsin (WI), animal studies, reproductive health, exposure assessment, biomedical research, heavy metalsRelevant Websites:
http://www.cvm.uiuc.edu/vb/friends_center/ Exit
Progress and Final Reports:
Original AbstractMain Center Abstract and Reports:
R829390 Water Innovation Network for Sustainable Small Systems Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
R829390C001 Neurobehavioral Effects of PCBs and Methylmercury in Rats
R829390C002 Perinatal PCB Exposure and Neuropsychological/Auditory Function
R829390C003 FRIENDS Analytical Toxicology Core Facility
R829390C004 Developmental Effects of PCBs and Methylmercury
The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.