Source: PURDUE UNIVERSITY submitted to
DEVELOPMENT OF METHODS OF ANALYSIS FOR FEEDS, FERTILIZERS AND PESTICIDES
Sponsoring Institution
State Agricultural Experiment Station
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
0095718
Grant No.
(N/A)
Project No.
IND053054S
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Oct 1, 2000
Project End Date
Sep 30, 2005
Grant Year
(N/A)
Project Director
Hanks, A. R.
Recipient Organization
PURDUE UNIVERSITY
(N/A)
WEST LAFAYETTE,IN 47907
Performing Department
STATE CHEMIST
Non Technical Summary
(N/A)
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
90172992000100%
Goals / Objectives
1. Develop methods for analysis of pesticides in formulated products of varied sample matrix. 2. Develop methods to extract phenoxy acid herbicides from soil and vegetation. 3. Develop alternate methods of analysis for antibiotics and drugs. 4. Develop and validate modified Kjeldahl for calcium and phosphorous in feeds. 5. Develop methods for vitamin A, lactose and other sugars, methoprene, vitamin D, melengestrol acitate, lasalocid and decoquinate in feeds.
Project Methods
1. Apply various extraction procedures in conjunction with liquid and gas chromatography to formulations. 2. Optimize parameters for use of "Accelerated Solvent Extraction". 3. Employ solid phase columns to separate mixtures and ELISA for difficult to analyze products like bacitracin. 4. Simplify and modify Kjeldahl digestion parameters and optimize for calcium and phosphorous in animal feeds sans rigor required for protein in same digest; and analyze Ca & P by interfaced, automatic colorimetric instrumentation. 5. Apply combinations of liquid chromatography, solid phase extraction and accelerated solvent extraction using appropriate detection systems for analyte determination.

Progress 10/01/00 to 09/30/05

Outputs
The corrections needed to calculate antibiotic values was investigated. Because of variations among standard reference zone readings, it is necessary to correct by plotting the log of the concentration versus the zone diameter to obtain a correction point. Further experiments will be conducted to determine all the factors that influence estimation of potency. Work continues on an LC method for Vitamin A. Development of an LC method for sugars in animal feeds is progressing. There is work being done on development of a quantitative method for chromium tripicolinate. Currently there is no method. Using ion chromotography, the laboratory is developing a method for choline chloride. The current method is gravimetric and non-specific. Improvements have been made to the taurine method, resulting in shorter assay time and improved separations. The laboratory is participating in collaborative studies for the drugs lasalocid sodium and oxytetracycline. This is in addition to the collaborative study for zearalenone, which is a mycotoxin. A method for non-nutritive metals in fertilizers, employing microwave digestion and ICP-OES instrument detection has been finalized, and an AOAC collaborative study is near completion. A formal report of the collaborative study, Analysis of Arsenic, Cadmium, Cobalt, Chromium, Lead, Molybdenum, Nickel, and Selenium in Fertilizers by Microwave Digestion and ICP-OES Detection: Collaborative Study, is being prepared for submission to JAOAC. A publication, Development of a Method to Determine Arsenic, Cadmium, Cobalt, Chromium, Lead, Molybdenum, Nickel, and Selenium in Fertilizers, describing the background development work establishing the collaborative method, has been prepared, and is in process of submission to JAOAC.

Impacts
New, faster, safer and more efficient analytical methods affords regulatory and industry laboratories opportunities for less-costly compliance assurance.

Publications

  • No publications reported this period


Progress 10/01/04 to 09/29/05

Outputs
The tylosin-urea adduct is a common component of cattle feeds. There are some problems with the assay as presented by Eli-Lilly (GP 7038). Long incubation periods are required and there is uneven diffusion in the plate assay. Adjusting the methanol buffer content to 20% improves the method performance. Bacitracin precision continues to be a problem. It may require a dilution of the sample to a specific reference concentration on the standard curve. More work is required. The OISC Laboratory successfully set up the PCR technique for determining prohibited material in animal feeds. The laboratory participated in an interlaboratory study with FDA to validate the method. An LC method for Vitamin A is being validated and an AOAC collaborative study is anticipated. Work on an LC method for Vitamin D in feeds is in progress. A more specific method for determining sugars as sucrose, glucose and fructose in feed products has been developed and is in the process of being validated. Other work in this area includes the determination of specific fatty acids and chromium tripicolinate. A method has been finalized for the determination of non-nutritive metals in fertilizer. It uses microwave digestion and determination by ICP-OES. Nutritive metals can also be screened at the same time. Selenium in animal feeds can also be routinely analyzed by the same procedure. The laboratory is looking into the possibility of transferring the phosphorus and potassium assays to ICP-OES. This will simplify the laboratory operation and enable us to remove some dated continuous flow equipment. Existing analytical methods for pesticide formulations were adapted for use in the OISC laboratory and evaluated for usefulness in regulatory enforcement. The following methods were adapted as satisfactory enforcement methods: Linalool by GC-FID, Nylar by GC-FID in a shampoo matrix, Phenols by LC-PDA in a towelette/wipe product matrix, Rotenone by LC-PDA in a shampoo matrix, Permethrin in plastic ear tags, Sulfluramid in cardboard baits, Sodium bromide by LC-IC in a large tablet and Naled in a liquid matrix. A method for Stabilene by LC-RI (refractive index detection) was found unsuitable for regulatory enforcement.

Impacts
New, faster, safer and more efficient analytical methods affords regulatory and industry laboratories opportunities for less costly compliance assurance.

Publications

  • No publications reported this period


Progress 10/01/02 to 09/30/03

Outputs
Bacitracin has multicomponent forms. Data shows sample diluation exerts a significant effect on results. The overall potency of the sample may show a negative or positive bias in results. Another factor is that different batches of feed grade bacitracin may vary in composition. Experiments are being conducted to determine what is the optimum dilution range that gives more valid results. Sample load has been high in the formulation laboratory during the last twelve months. The chemists have been fully occupied with routine analysis of official OISC samples and region 5 EPA backup Lab samples. A liquid chromatographic method for the determination of the drug, sulfathiazole, in swine feeds was developed, validated, and published (J. AOAC Int.) A liquid chromatographic method for the determination of riboflavin in feeds was developed, validated, and published (J. AOAC Int.) Work has begun on a liquid chromatographic method for determination of fatty acids in animal feeds, including linoleic acid, linolenic acid, arachidonic acid, omega-3 fatty acids, and omega-6 fatty acids. Work has continued on a liquid chromatographic method for the determination of vitamin D3 in feeds which employs solid-phase extraction for sample cleanup. Validation work is continuing on a liquid chromatographic method for determination of vitamin A in animal feeds. Work has started on the development of a method for determination of choline chloride in animal feed using ion chromatography. Development has resumed on a method for determination of chromium as chromium tripicolinate in animal feed at typical use levels (200 ppb). Non nutritive metals in fertilizers remains a priority regulatory concern. Using microwave digestion procedures previously developed, ICP-OES instrument procedures have been refined. This laboratory is currently collaborating with other state regulatory and fertilizer industry laboratories in establishing a rigorous instrument procedure which will be applicable to various ICP-OES instrument designs and models. A study is also underway to verify fertilizer ICP-OES instrument results relative to ICP-MS results. For nutritive metals in fertilizers, ICP-OES instrumentation has been successfully validated relative to traditional Flame Atomic Absorption instrumentation. For selenium in animal feeds, a new procedure has been developed using microwave digestion of samples, and ICP-OES analysis. The procedure has been favorably compared to a more laborious manual digestion procedure, followed by hydride generation Flame Atomic Absorption.

Impacts
New, faster, safer and more efficient analytical methods affords regulatory and industry laboratories opportunities for less costly compliance assurance.

Publications

  • Britton, N.L., Riter, K.L., Smallidge, R.L., Hillebrandt, J. 2002. Reverse-phase liquid chromatographic determination of riboflavin in feeds. J. AOAC Int., 86 (2), 197-201.
  • Albert, K., Riter, K.L., Smallidge, R.L. 2002. Determination of sulfathiazole in swine feed by reverse- phase liquid chromatography with post-column derivatization. J. AOAC Int., 86 (4), 623-630.
  • Newlon, N., Comparison of the Ammonium Oxalate Extraction of Fertilizer for K20, Method 983.02, with the Ammonium Citrate/EDTA Extraction, Method 993.31, Journal of AOAC International, Vol 86, Nos 4, 2003, p. 640-642.


Progress 10/01/01 to 09/30/02

Outputs
The analysis of heavy metals in fertilizers continues to be a regulatory priority. An inter-laboratory study was conducted to evaluate various acid digestions on preparing fertilizer materials for analysis of heavy metals. A hot plate and microwave procedure have been selected for use with ICP-MS and ICP-OES detection. An ICP instrument has been ordered for the OISC Laboratory to be used for this purpose. A method to determine sulfathiazole in animal feeds has been developed and submitted for publication in the JAOAC. Another drug method, Tiamulin in Feed, has been published in JAOAC. Progress has been made on LC methods for the determination of total sugars and lactose in feeds. This makes the determination much more specific and accurate. Work on Vitamin D3, Biotin and Vitamin A methods of analysis continue with great progress being made. An antibiotic in feed, Monensin, has a very lengthy incubation period as part of its assay. Work is progressing to limit the incubation time to about half of the current seven hours. The antibiotic method for Bambermycin can give erratic results at times. Experiments are being conducted to determine the cause. In the pesticide formulation laboratory there are several active ingredients being investigated. 2,2-dibromo-3-nitrilopropionamide (DBNPA) is found in antimicrobial products and a method using normal phase LC is being developed. In addition, a spectrophotometric method for boron in wood stakes, an ion chromatography method for sodium bromide are also being developed. Propylene glycol and rabon in feeds are two pesticide feed ingredients for which GC-FID methods are being developed. In paint products, a method for methylene bis isothiocyanate (MBTC) and chlorothalonil which is a contaminant are being looked at by use of LC. Finally, a GC-ECD method for contaminant permethrin in methoprene in formulated pet insecticidal products is being developed.

Impacts
New, faster, safer and more efficient analytical methods affords regulatory and industry laboratories opportunities for less costly compliance assurance.

Publications

  • No publications reported this period


Progress 10/01/00 to 09/30/01

Outputs
The importance of heavy metal analysis has increased recently. A method survey study was conducted to determine method variations. Development of a standardized method using microwave digestion is being pursued. The analysis will be by high resolution ICP-MS. An improved HPLC method for analysis of the drug tiamulin was developed. This method allows for increased sample throughput, decreased solvent useage and improved chromatography. A new method for the determination of lactose in animal feeds was developed using liquid chromatography with a refractive index detector. It allows the analysis to be done in the presence of whey which is not possible by the current official method. Improvements were made in the liquid chromatography method for carbadox and pyrantel tartrate drugs. The method increases sensitivity and reproducibility of the results. Vitamin A method efficiency was improved by adding new dispensers and automatic pipettes. Analysis by chromatography is being improved to reduce analysis time. The effect of methanol concentration, pH and feed concentration is being studied for two different strains of M. Luteus which is used in the assay of bacitracin in animal feeds. An analytical method was developed for ammoniated salts of fatty acids which are used as home use weed killers. These are examples of the newer safe pesticides using natural ingredients. The method uses methylation of acids followed by extraction and determination by gas chromatography. A new method is being developed to determine individual specific quaternary ammonion compounds. These are used as sanitizers and cleaning agents in hospitals, schools and homes. Current methodology does not allow the determination of mixtures of these compounds. The approach that will be followed is suppressed ion conductivity detection with ion pairing.

Impacts
New, faster, safer and more efficient analytical methods affords regulatory and industry laboratories opportunities for less costly compliance assurance.

Publications

  • Hanks, A.R. 2001. General Referee Reports, Pesticide Formulations: CIPAC Studies, J AOAC
  • Newlon, N. 2001. Comparison of Ammonium Oxalate Extraction of Fertilizer for K20, Method 983.02, with Ammonium Citrate/EDTA Extraction, Method 993.31
  • Kane, P. 2001. Regulation of Heavy Metals in Fertilizer: The Current State of Analytical Methodology, ACS
  • Kane, P. 2001. General Referee Report, Fertilizers and Agricultural Liming Materials, JAOAC
  • Moore, D.B., Britton, N.L., Smallidge, R.L., Riter, K.L. 2001. Determination of Tiamulin in Type C Medicated Swine Feeds Using High Throughput Extraction with Liquid Chromatography. J. AOAC Int'l
  • Lin, H., Riter K. 2001. Improved Chromatography for the HPLC Determination of Carbadox and Pyrantel Tartrate. Forum on Methods for Antibiotics and Drugs in Feeds. AOAC Int'l 115th Annual Meeting
  • Moore, D.B., Britton, N.L., Smallidge, R.L., Riter K.L. 2001. Further Development and Validation of a Method for the Determination of Tiamulin Type C Medicated Swine Feed Using HPLC. Forum on Methods for Antibiotics and Drugs in Feeds. AOAC Int'l 115th Annual Meeting


Progress 10/01/99 to 09/30/00

Outputs
Analytical progress has been reported for determination of calcium and phosphorous in feeds. Shifting protein analysis to combustion methodology has allowed use of less rigorous Kjeldahl block digestion while continuing the use of autoanalyzer methodology for analysis of calcium and phosphorous. Using a lower digestion block temperature, decreased salt/acid ratio and decreased digestion time, results were comparable to calcium by atomic absorption and gravimetric phosphorous, while essentially using a safer, faster and more efficient method. Methoprene is used in feeds as a pass through growth regulator to control horn flies in manure. A method of analysis for methoprene in feeds has been developed using hexane extraction, Florisil solid phase cartridge cleanup, followed by reversed phase HPLC and quantitation by UV absorption. The method is being optimized and subjected to within laboratory validation. The effect of pH has been explored and compared for impact on the determination of tylosin by both plate and turbometric, microbiological methods. Tylosin B activity varied with pH and method of analysis. Other experiments revealed difficulty in obtaining valid results for tylosin when tylosin B exceeds 20%. Modification to the tylosin method has been made to eliminate the positive bias. Work has begun on trace metals in fertilizers using different instrumental methods combined with various test sample digestion alternatives. Progress continues on method development for numerous other drugs, vitamins, pesticides, and total sugars and lactose.

Impacts
New, faster, safer and more efficient analytical methods affords regulatory and industry laboratories opportunities for less costly compliance assurance.

Publications

  • Ragheb, H.S. 2000. General Referee Report: Antibiotics in feeds. J. AOAC Int'l 83, 503-505
  • Kane, P.F. 2000. General Referee Report: Fertilizers and agriculture liming materials. J. AOAC Int'l 83, 509-510
  • Hanks, A.R. 2000. General Referee Report: CIPAC studies. J. AOAC Int'l 83, 431-433
  • Patel, R. and Kane, P. 2000. New, safer and more efficient block digestion procedure for the determination of calcium and phosphorus in animal feeds by autoanalyzer. Abstract. AOAC Int'l 114th Annual Meeting
  • Moore, D.B., Britton, N.L., Smallidge, R.L. and Riter, K.L. 2000. Determination of tiamulin in type C medicated swine feeds using small-scale extraction with improved HPLC analysis. Forum on methods for antibiotics and drugs in feeds. AOAC Int'l 114th Annual Meeting
  • Newlon, N.F. 2000. Evaluation of AOAC performance parameters of K2O in fertilizers using method 983.02 with new autoanalyzer instrumentation. Abstract. AOAC Int'l 114th Annual Meeting
  • Moore, D.B. and Riter, K.L. 2000. Determination of Methoprene in cattle feeds using solid phase extraction with HPLC analysis. Abstract. AOAC Int'l 114th Annual Meeting
  • Newlon, N.F. and Kane, P.F. 2000. Improved autoanalyzer instrumentation for P2O5 in fertilizers using method 978.01: consistent and reliable results, trials and tribulations. Abstract. AOAC Int'l 114th Annual Meeting
  • Ragheb, H.S. and Troyan, T.A. 2000. Further studies on the microbiological assay of Tylosin and TUA in feeds. Forum on methods for antibiotics and drugs in feeds. AOAC Int'l 114th Annual Meeting


Progress 10/01/98 to 09/30/99

Outputs
Progress continues on changes to an automated instrument systems for the analysis of P2O5 and K2O in fertilizers. Work to validate these methods relative to existing standard methods is near completion. Newly modified sample digestion for Ca and P in feeds is encouraging for use in automated analysis of these mineral nutrients. Comparison data of the new digestion process and instrument systems is being generated. Non-nutritive (heavy metal) analyses have been developed covering Pb, Cd and As for possible validation in analysis of both feeds and fertilizer. Validation of a HPLC post column derivatization method for sulfamethazine in feeds has been completed while studies on sulfathiazole are near completion. Progress continues on methods for decoquinate and lasalocid where work includes overcoming difficulties with mineral feeds. A method study for riboflavin is near completion and developmental efforts for vitamin D3 have managed the elimination of CHCl3 in the mobile phase. The tylosin-urea adduct formed in feed must be hydrolyzed prior to assay. It has been found autoclaving was adequate for hydrolysis. Clean up of the extract on acidic alumina works for the plate assay while a pH adjustment will be needed for the turbidimetric assay of the extract after clean-up. Methods of analysis for feeds, fertilizers and pesticides are important for regulation of these agricultural production inputs to assure consumers receive what is claimed on product labels, provide a fair environment for trade, and to protect health and the environment. Methods have been developed for the analysis of vitamin A and the drug decoquinate in animal feeds using less solvent and toxic chemicals. Progress continues on rapid microwave digestion of feeds and fertilizers for analysis of important trace mineral nutrients using coupled online dilution and automated atomic absorption. Assay interferences of various drugs and antibiotics have been further defined.

Impacts
Faster and automated chemical analysis reduces cost per analysis while reduction of solvent and toxic chemical use reduces human exposure and costly disposal of toxic waste. These results add to economical analysis of samples in less time and more efficient regulatory programs protecting health, safety and preventing consumer fraud.

Publications

  • Ragheb, H.S. 1999. General Referee Report. Antibiotics in feeds. J. AOAC Int=l 82, 447-448
  • Kane, P.F. 1999. General Referee Report. Fertilizers and agricultural liming materials. J. AOAC Int=l 82, 453-454
  • Hanks, A.R. 1999. General Referee Report. Pesticide formulations: CIPAC studies. J. AOAC Int=l 82, 496-499


Progress 10/01/97 to 09/30/98

Outputs
New work on the analysis of vitamin A in animal feedstuffs by HPLC is progressing well and yields three advantages: avoids the use of disposal of toxic chemicals (chloroform and antimony trichloride) required by the traditional calorimetric method, increased sample throughout/productivity over the calorimetric method, and decreased use and disposal of solvents over other available liquid chromatographic methods. Efforts continue on the HPLC analysis of decoquinate in feed using fluorescence detection. Cation coextractives suspected of influencing both recovery and fluorescence are being examined. Experiments to test potential interferences by coextractive drugs in feeds with the hygromycin assay indicates amprolium and bacitracin do not cause interference, while sodium chloride can be antagonistic. Further testing of salt, penicillin and chlorotetracycline which appear to be removed by cation exchange column cleanup, is in process. Automated microwave digestion in place of a manual hotplate approach has been found effective for fertilizers, but not for feeds, in trace metal analysis by atomic absorption coupled with automatic online digest dilution and aspiration. Instrument modification appears needed to accommodate feeds and a new modified bata instrument is expected soon. Testing and perfecting of methods for pesticides in ground water have been completed: solid phase extraction of herbicides and insecticides with analysis by GC/MSD, analysis of phenoxy herbicides by HPLC and analysis of carbamates by HPLC with post-column derivatization. Plans include modification of HPLC procedures to permit analysis by LC/MSD in the future.

Impacts
(N/A)

Publications

  • Kane, P. F. and Newlon, N. F. 1997. A comparison of continuous flow microwave digestion vs traditional hot plate digestion of fertilizers and animal feeds for trace element analysis by atomic absorption. Abstract. AOAC Int'l Central Section 10th Annual Meeting
  • Hanks, A. R., 1998. Liquid chromatographic method for determining thiodicarb in technical products andformulations: CIPAC collaborative study. J. AOAC Int'l 81, 341-343
  • Ragheb, H. S. 1998. General Referee Report. Antibiotics in feeds. J. AOAC Int'l 81, 203-204
  • Smallidge, R. L. 1998. General Referee Report. Drugs in feeds. J. AOAC Intl 81, 204-207
  • Kane, P. F. 1998. General Referee Report. Fertilizers and agricultural liming materials. J. AOAC Int'l 81, 211-212.
  • Hanks, A. R. 1998. General Referee Report. Pesticide formulations: CIPAC studies. J. AOAC Int'l 81, 115-117
  • Ragheb, H. S., Herrick, C. A. and Miles, D. A. 1997. Microbiological assay of tylosin in combination with monensin in feeds. Abstract. AOAC Int'l Central Section 10th Annual Meeting
  • Reeve, J. S. and Smallidge, R. L. 1997. Aerosol sampling in pesticide formulations. Abstract. AOAC Int'l Central Section 10th Annual Meeting


Progress 10/01/96 to 09/30/97

Outputs
A method for pesticide residues in soils and vegetation has been developed using accelerated solvent extraction - solid sample extraction using pressurized solvents. Semi-automated extraction of pesticide residues from water with recovery on solid phase extraction disks has been developed for use in a baseline study of pesticides in ground water. These extraction methods reduce both solvent and personnel time, yet maintain or improve extraction capabilities. Work is in progress to develop automated microwave digestion procedures to replace the current manual hotplate approach to digestion. Coupled with automated, online instrumental dilution of digests, this process will provide a completely automated sequence for sample analysis after weighing of samples, resulting in significant saving of hands-on personnel time. Sampling from pesticide aerosol products has been investigated for two types of aerosols by two different subsampling approaches. Both single phase aerosols (organic) and dual phase aerosols (aqueous/organic) have been sampled by spraying off the top and acetone/dry ice freezing followed by total propellent exhaustion. While organic based products appear amenable to both sampling approaches, mixed phase products need to be subjected to the freeze-exhaust approach. Chlorotetracycline and monensin have been found to interfere with assay of bacitracin in feeds at low contentrations of bacitracin. At high levels of bacitracin, interference effects are minimized at greater dilution of extracts before analysis. For turbidimetric assays, but not diffusion plates, acidic alumina has been found to eliminate interference from monensin in the assay for tylosin.

Impacts
(N/A)

Publications

  • Ragheb, H. S. 1997. General Referee Report. Antibiotics in Feeds. J. AOAC Intl 80, 184-185.
  • Smallidge, R. L. 1997. General Referee Report. Drugs in Feeds. J. AOAE Intl 80, 188-189.
  • Riter, K. L. 1997. Troubleshooting Analytical Methods: Estimating Sources of Error. Proc. AOAC Antibiotic and Drug Forum. 111th AOAC Intl Annual Meeting.
  • Herrick, C. A., Miles, D. A. and Ragheb, H. S. 1997. Amprolium Progress Report. Proc. AOAC Antibiotic and Drug Forum. 111th AOAC Intl Annual Meeting.
  • Ragheb, H. S., Herrick, C. A. and Miles, D. A. 1997. Assay Interferences by Some Antibiotic Combinations. Proc. AOAC Intl Antibiotic and Drug Forum. 111th AOAC Intl Annual Meeting.
  • Hanks, A. R. 1997. Liquid Chromatographic Method for Determination of Thiodicarb in Technical Products and Formulations: CIPAC Collaborative Study. J. AOAC Intl 80, in press.
  • Kane, P. F. 1997. General Referee Report. Fertilizers and Liming Materials. J. AOAC Intl 80, 186-187.
  • Hanks, A. R. 1997. General Referee Report. CIPAC Studies. J. AOAC Intl 80, 107-109.


Progress 10/01/95 to 09/30/96

Outputs
A method for taurine in cat food has been developed. After automated precolumn derivatization, it is separated from other amino acids using reversed phase HPLC. Complete validation of the method should be achieved by late Spring of 1997. Work is underway for vitamin A analysis by HPLC. Efforts are for improved extraction and recovery agreeing with standard methods. HPLC analysis of vitamin A greatly reduces solvent wastes. Further solvent savings are sought through solid phase extraction (SPE). Other HPLC methods are being developed for vitamin E and decoquinate in feeds and phenoxy acids in fertilizers. Enzyme linked immuno assays are being explored in analysis of hygromycin and tylosin in feeds using this technique in a quantitative mode, while SPE is being found to improve recovery and precision in analysis of apromycin. Combustion analysis of sulfur in fertilizer is proving to be accurate while reducing labor over standard methods. Publication and collaboration study validation of the method are planned. Research on automated microwave digestion preparation of feeds and fertilizers for trace element analysis is in process. Linkage to automated, online instrumentation for dilution and sample introduction in atomic absorption analysis is planned for greater system efficiency. HPLC analysis of 12 phenoxy herbicide trace residues in ground water has been validated with an added bonus of reduced analyst exposure to chemical hazards. SPE is being explored to reduce solvent use and extraction time.

Impacts
(N/A)

Publications

  • Kane, P.F. and Newlon, N.F. 1995. Improved precision in the automated analysis of P2O5 in fertilizer. J. AOAC Int'l. 78:110-114.
  • Kane, P.F. 1996. General referee report. Fertilizer and agricultural liming materials. J. AOAC Int'l. 79:267-268.
  • Ragheb, H.S. 1996. General referee report. Antibiotics in feed. J. AOAC Int'l. 79:264.
  • Ragheb, H.S. 1996. Apromycin progress report. Proc. AOAC Antibiotic and Drug Forum. 110th AOAC Int'l. Annual Meeting.
  • Smallidge, R.L. 1996. A discussion of standard addition and internal standard techniques as used in analysis of drugs in feeds. Proc. ISO/IUPAC/AOAC Symposium - Use of recovery factors.
  • Smallidge, R.L. 1996. Summary guidelines for significant figure determination. Proc. AOAC Antibiotic and Drug Forum. 110th AOAC Int'l. Annual Meeting.
  • Smallidge, R.L. 1996. General referee report. Drugs in feeds. J. AOAC Int'l. 79:266-267.
  • Thiex, N., Smallidge, R.L. and Beine, R. 1996. General comments on sources of error in vitamin A analysis. J. AOAC Int'l., in press.
  • Hanks, A.R. 1995. Liquid chromatographic method for determination of dithianon in technical products and formulations: CIPAC collaborative study. J. AOAC Int'l. 78:1131-1133.


Progress 10/01/94 to 09/30/95

Outputs
A collaborative study has been initiated for the study of a method covering sulfamethazine in swine and cattle feeds. The method was developed in this laboratory and perfected over the last few years. It is more specific, accurate, sensitive and precise than currently accepted methods for sulfamethazine. Rapid solid phase extraction methodologies for pesticide residues in water and soils have been developed and are under further evaluation and characterization. Clear advantages in sample preparation time are evident while solvent savings provide both economic and environmental benefits. The laboratory has become a beta test site for new atomic absorption equipment and doing evaluations of software and automated sample dilution and introduction hardware. The new atomic absorption systems are allowing the discovery of inadequacies in existing methodologies for plant and animal, mineral nutrients in fertilizers and feeds. Work continues on improving precision in the automated analysis of P2O5 and on hydride generation atomic absorption for analysis of selenium as a replacement for current wet chemistry. Steps in the assay of apramycin in feeds are being examined as the source of large variations in this analysis. Minor variations in sample volumes used for zone inhibition analysis have been found to be a source of error and use of ion exchange is being examined for elimination.

Impacts
(N/A)

Publications


    Progress 10/01/93 to 09/30/94

    Outputs
    A collaborative study has been initiated for the study of a method covering sulfamethazine in swine and cattle feeds. The method was developed in this laboratory and perfected over the last few years. It is more specific, accurate, sensitive and precise than currently accepted methods for sulfamethazine. Rapid solid phase extraction methodologies for pesticide residues in water and soils have been developed and are under further evaluation and characterization. Clear advantages in sample preparation time are evident while solvent savings provide both economic and environmental benefits. The laboratory has become a beta test site for new atomic absorption equipment and doing evaluations of software and automated sample dilution and introduction hardware. The new atomic absorption systems are allowing the discovery of inadequacies in existing methodologies for plant and animal, mineral nutrients in fertilizers and feeds. Work continues on improving precision in the automated analysis of P(subscript 2)O(subscript 5) and on hydride generation atomic absorption for analysis of selenium as a replacement for current wet chemistry. Steps in the assay of apramycin in feeds are being examined as the source of large variations in this analysis. Minor variations in sample volumes used for zone inhibition analysis have been found to be a source of error and use of ion exchange is being examined for elimination.

    Impacts
    (N/A)

    Publications


      Progress 10/01/92 to 09/30/93

      Outputs
      Autoanalyzers (Traacs) have been reconfigured from original instrumentation to maximize precision in determining phosphate in fertilizers. Tube pump platens were machined to higher tolerances to improve performance and sampler wash reagent modified to match the sample matrix. Insulated manifold tubing was used to reduce heating and cooling effects. The combination of changes results in a 66% reduction of the standard deviation for replicate analyses. Fluorometric methods (for food and plant materials), a graphite furnace atomic absorption spectraphotometric method, and a hydride generation method we developed are being compared for the analysis of selenium in animal feed premixes. Research continues on a method for apromycin in feeds with the goals of understanding the role of cations in analytical sensitivity and the elimination of the current ion exchange clean up prior to analysis. An HPLC method for three common herbicides (2,4-D, MCPP and dicamba) is being refined for the analysis of weed and feed lawn fertilizers. Methanol/acetic acid/water as well as methanol/citrate buffer mobile phases show great promise with an amino column. Solid phase extraction is being applied to recovery of pesticides extracted from soils. Chemically bonded C-18 silica enmeshed in an inert matrix of polytetrafluorethylene in disk form appears well suited for a faster, reduced solvent, less labor intensive extraction/pesticide recovery step in sample preparation.

      Impacts
      (N/A)

      Publications


        Progress 10/01/91 to 09/30/92

        Outputs
        Study of a method for analysis of the antibiotic apramycin in animal feeds indicates a need for improved precision. Progress is being made and we anticipate completion and publication of this work soon. The cup-agar method of assay for neomycin developed in this lab has been compared to the standard cylinder method in a multiple laboratory collaborative study in which this laboratory participated. In this study the cup-agar method proved superior to the cylinder method. 2,4-Dichlorophenoxy acetic acid analysis using separation on an amino liquid chromatographic column in the ion exchange mode provides better resolution of several components not commonly resolved with the standard reversed phase method, especially with low level 2,4-D formulations and use dilution fertilizers. This approach should lead to better analysis of this product. Efficacy testing of sporicidal disinfectants has been completed and methods for tuberculocidal products are being initiated. The need for improved methods is obvious. Intensive wet chemistry methodology is being replaced by an instrumental approach for the determination of selenium in animal feeds. Graphite furnace atomic absorption is being explored. Analysis of the mycotoxin fumonosin is being automated. Use of automation for the o-phthaladehyde derivatization and liquid chromatographic analysis of this mycotoxin is proving successful. The laboratory should be in position to rapidly respond to future demands for the analysis of this toxin.

        Impacts
        (N/A)

        Publications


          Progress 10/01/90 to 09/30/91

          Outputs
          A method for the determination of the antibiotic neomycin in animal feeds has been examined for sources of variation. The majority of the method variation has been found associated with variation between plates. By using agar wells cut in the plates in place of stainless steel cylinders for adding feed extracts to plates, precision is significantly improved. This technique is being readied for publication. The previously developed double extraction procedure for the antibiotic bacitracin has been explored through a multi-laboratory study. Data is still being evaluated. A fertilizer nitrogen method using combustion technology and equipment has been finalized and is ready for multi-laboratory testing to determine within and between laboratory variation. Flame photometric analysis of potassium in fertilizers is being updated and validated with lab on new, third generation, automated equipment. Similar efforts for fertilizer phosphorous analysis is also being advanced for collaborative study of performance in several laboratories. Post column derivatization (PCD) following HPLC is used in the analysis of sulfa drugs in animal feeds. Preliminary studies on an HPLC-PCD method have been completed in preparation for analysis of samples in several laboratories. This same methodology is being compared with commercial immuno-assay kits. Results show some promise that the kit could be used for screening, or even confirmation, in the analysis for contamination of sulfas in feeds.

          Impacts
          (N/A)

          Publications


            Progress 10/01/89 to 09/30/90

            Outputs
            Testing of methods for nitrogen in fertilizers continues with a variety of instrumentation. Combustion technology gives good results for solid fertilizers, but changes are required for analysis of solutions. Extractant modification is being explored in the analysis of sulfathiazole and sulfamethazine residues in feeds with detection by post-column derivatization after HPLC. Similar final determination is being applied to feeding levels with method enhancement for sulfathiazole using sulfamethylthiazole, provided by the FDA, as an internal standard. Plans are being made to pursue the negative bias found in the analysis of feeds of lasalocid and pyrantel tartrate through multiple standard addition experiments. Extraction of the antibiotic ardacin from feeds is difficult and requires a strong polar solvent. A mix of solvents and buffer and varying pH has been tried. Extraction is found to be good with 20% acetonitrile: 20% dimethyl sulfoxide: 60% pH 8 buffer. While successfully applied to premixes and concentrates, at complete feed levels, it is necessary to minimize interferences and research is so directed. Classically, the analysis of bacitracin MD in feeds has presented problems. Recovery improves on complete feeds with double extraction. Variation in plate assay zone diameters of inhibition remains a problem targeted for further research.

            Impacts
            (N/A)

            Publications


              Progress 10/01/88 to 09/30/89

              Outputs
              Pesticides: methodology for nitrate and nitrite is being investigated for application to both pesticides and animal feeds. Fertilizers: The traditional Kjeldahl nitrogen method is both resource and labor intensive and requires a large amount of operational space. In addition, the current method presents several environmental drawbacks. Investigation of a replacement method is underway. Feeds: (drugs, vitamins and amino acids) - Work continues toward improving methods for analysis of fat soluble vitamins D and E by HPLC. An interlaboratory study is being prepared on a post-column reaction method for determining sulfa drugs. Feeds: (antibiotics) - The glycopeptide antibiotic, ardacin, is added to feeds at low levels (5-10 ppm). Affinity chromatography is being explored to eliminate co-extraction interferences in analysis. Feeds: (other) - Studies are underway to evaluate the heavy metal contents of various mineral feed ingredients, especially phosphate sources. Atomic absorption with graphite furnace techniques is being used in these studies.

              Impacts
              (N/A)

              Publications


                Progress 10/01/87 to 09/30/88

                Outputs
                Pesticides (formulations): Work has been completed on the development of a HPLC method providing for quantitative analysis of organic arsenical herbicides. The method gives greater specificity in the analysis of this class of pesticides than previously available. A method for separation of the herbicides benefin and trifluratin has also been perfected and a method for fenoxapropethyl (Whip) analysis validated. Pesticides (residues): Work continues on the comparison of immunoassay and gas chromatograph for the determination of chlordane in soils prior to an interlaboratory study for method validation. Feeds (drugs, vitamins and amino acids): Vitamin D methodology continues in development along with vitamin E, while the analysis of vitamin A by HPLC is still being pursued. Although a residue method for sulfa drugs has been developed and reported, work is ongoing for further improvement. Feeds (antibiotics): cygro and stenerol have been shown to not interfere significantly in the analysis of virginiamycin, while the separation and assay of virginiamycin and monensin have been achieved using solid phase extraction. Feeds (other): A collaborative study is underway to validate feed protein analysis using CuSO(subscript 4), catalyst and modified block digestor conditions. Fertilizers: Combustion techniques and equipment are being studied for the analysis of nitrogen. Success has been achieved with solids while liquids and suspensions require additional research.

                Impacts
                (N/A)

                Publications


                  Progress 10/01/86 to 09/30/87

                  Outputs
                  Pesticides (formulations): methods for four new herbicide formulations (Reflex,Banner, Lorox Plus and Tendem) have been developed. Boron and ammonium sulfamate methods using ion-exchange chromatography have been completed, while the determination of organic arsenicals by LC is underway. Pesticides (residues): various chlordane in soil methods are being reviewed for us in an AOAC collaboration study. LC with post column fluorescence detection is being examined for glyphosate and N-methyl carbamates. Feeds (antibiotics): analysis of virginiamycin in feeds containing cygro or stenerol is in the planning stage. Reasons for the observed depression of virginiamycin activity by monensin in the plate assay are also to be investigated. Feeds (drugs, vitamins and amino acids): methods for the determination of sulfa drug residues and amprolium in feeds have been completed. Method development for lysine analysis is nearing completion, while method work for vitamin D and riboflavin is underway. Feeds (other): Iodide, iodate and periodate by LC is being developed and a mercury free Kjeldahl method for protein is ready for collaborative study. Third generation automation equipment is being fine tuned for the analysis of protein, calcium and phosphorus. Fertilizers: extensive experience with efforts to determine nitrogen by block digestion is being described for publication. The new automation equipment is also being described for publication.

                  Impacts
                  (N/A)

                  Publications


                    Progress 09/01/85 to 10/30/86

                    Outputs
                    Pesticides: routine gas chromatographic methods have been developed for cypermethrin, fenitrothion, famphur, hydroprene and parinol. Seven pesticide methods have been adopted to megabore column gas chromatography and low level pyrethrin analysis developed on capillary. Liquid chromatograph has been applied to the analysis of six pesticides. An assay for cholecalciferol is being prepared for publication. Feeds (antibiotics): an ion exchange separation has been applied to the analysis of feeds containing both chlortetracycline and virginiamycin. A manuscript on ion exchange removal of chlortetracycline prior to virginiamycin assay is in preparation. Feeds (drugs and vitamins): a finalized method for amprolium is being readied for publicaton while work continues on methods for lysine, lasalocid sodium, phenylarsonic acids, riboflavin and sulfa drugs. Methodology for sulfa drug residue analysis will soon be finalized. Feeds (other): a collaborative study has been completed comparing Hg0 and CuS0(4)/Ti0(2) as catalysts for determination of protein. Method development for the quantitation of iodine, iodate and periodate by liquid chromatography is underway. Fertilizer: a study to develop a method for determining nitrogen in fertilizer by use of a block digester and steam distillation has been started.

                    Impacts
                    (N/A)

                    Publications


                      Progress 01/01/85 to 09/30/85

                      Outputs
                      Pesticides: efforts to chromatographically resolve three closely related compounds (Trifluralin, Benfluration and Ethofluralin) are being made using capillary GLC and a variety of columns and solvent systems with HPLC. Feeds (antibiotics): the neomycin assay has been improved with a resultant increase in sensitivity and minimization of interferences. Elimination of lasalocid interferences in the determination of virginiamycin has been demonstrated using hexane partitioning, however, monensin and narasin interferences remain a problem. Analysis of lasalocid by HPLC is being found to compare favorably with the microbial method. Feeds (drugs & vitamins): a new method for amprolium analysis has been developed using CaCl(2) and an ion pairing reagent in the extractant. Addition of the ion pairing reagent to the HPLC solvent improved chromatographic performance. Vitamin D is found to be a difficult analyte for development of a comprehensive method. DMSO and DMF water base extractants work on some but not all feeds. Fat content of feeds effects the recovery by influence on the partition coefficient. Feed (other): a reverse phase HPLC method for lysine has been developed and found applicable to a variety of feeds. Significant time savings in sample digestion has been achieved with a mercury free catalyst, CuSO(4)-TiO(2), for protein analysis. Fertilizers: nitrogen analysis using a block digestor and steam distillation is being explored to save time and reagents.

                      Impacts
                      (N/A)

                      Publications