AU2005201412B2 - Method For Determining Biocides Within Wood - Google Patents

Method For Determining Biocides Within Wood Download PDF

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AU2005201412B2
AU2005201412B2 AU2005201412A AU2005201412A AU2005201412B2 AU 2005201412 B2 AU2005201412 B2 AU 2005201412B2 AU 2005201412 A AU2005201412 A AU 2005201412A AU 2005201412 A AU2005201412 A AU 2005201412A AU 2005201412 B2 AU2005201412 B2 AU 2005201412B2
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wood
solvent
extraction
filtration
extraction solvent
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AU2005201412A1 (en
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Peter James Hayward
Jarett Malcolm-Black
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Arxada NZ Ltd
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Arxada NZ Ltd
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  • Chemical And Physical Treatments For Wood And The Like (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Description

Title of the Invention Method of Determining Biocides Within Wood 5 Background to the Invention The invention relates to a method for determining biocides within wood. 10 Presently, chemical preservatives are added to wood, wood-based products and engineered wood products to protect them. The preservatives can include insecticides, fungicides, mouldicides and bacteriocides. For example, the preservatives can be insecticides to protect the products from attack 15 by insect species. In order to ensure that such wood-based materials are treated with sufficient insecticide to inhibit attack, it must be possible to quantify the amount of insecticide that is infused into the material. As is known in the art, the families of insecticide presently used are neonicotinoids, 20 synthetic pyrethroids, organophosphates and phenyl pyrazoles. As the potency of the compounds within these families has increased, the quantity of the compounds required for protection of timber related materials has decreased remarkably. The resultant low dosages have made it difficult to detect the active ingredients in the timber related materials, let alone quantify them. 25 1 Conventional methods of analysis for determination of the active content of wood based materials rely on chromatography, specifically gas chromatography (GC) and high performance liquid chromatography (HPLC). Although such chromatographic techniques yield excellent results for the analysis of the above mentioned insecticides in 5 water or sap extractions from vegetable based materials, they give reduced recoveries when applied to the analysis of preservatives such as insecticides in wood and wood based materials. This is especially true in the presence of plastic or resin based adhesives or coatings in or on the wood and wood-based materials. 10 Immunoassays such as enzyme linked immunosorbent assays are presently used for the analysis of pesticides on wood, vegetables and surface treated seeds. The use of immunoassay techniques using conventional immunoassay methodology in an effort to analyse the amount of insecticide infused into wood-based materials has revealed that these techniques are inadequate for infused insecticides due to the low recoveries. 15 The term "preservative" is understood to include not only the original active ingredient contained within the timber treatment formulation but also biologically active derivatives of the active ingredient that may arise during the timber treatment process or during the lifetime of the treated product. 20 2 Summary of the Invention It is therefore an object of the present invention to provide a method for determining biocides within wood wherein a high recovery is achieved. 5 Broadly in one aspect the invention consists of a method for quantitation of chemical preservatives used to treat wood, wood-based products and engineered wood products characterised in the use of an enzyme linked immunosorbent assay. 10 The present invention is based on the surprising discovery that when aspects of the immunoassay technology are combined with aspects of chromatography analyses when applied to the analysis of preservatives in wood and wood-based materials, a very high recovery is obtained. 15 In a preferred form of the invention high performance liquid chromatography extraction methodology is applied to an enzyme linked immunosorbent assay ("ELISA") technique. Surprisingly, the invention in this form yields high rates of extraction of imidacloprid relative to conventional methodology for both high performance liquid chromatography 20 and immunoassay techniques. 3 Description of the Preferred Embodiment of the Invention According to a preferred form of the invention the method includes the steps of reducing the particle size, through mechanical means of the wood, wood-based material 5 or engineered wood product to be extracted, using organic liquids that can penetrate the wood matrix as extraction solvents, filtration of extracts and preparation of standards using appropriate untreated wood extracts. Preferably the particle size of the wood-based material being analysed is reduced to 10 between 1000 and 10 micrometres diameter or less. The reduction in particle size can be carried out before extraction solvent using a Wiley mill or after extraction solvent is added using an industrial type high shear blender like a Silverson_. The extraction solvent is generally an organic solvent such as, but not restricted to, 15 methanol, dichloromethane, acetone, or toluene. Non water miscible solvents are then stripped from the extract by evaporation under reduced pressure and taken into either water or methanol prior to analysis with ELISA techniques. Water miscible solvents can be used providing they are diluted into water by a ratio of 400:1 or greater. 20 The filtration of extracts can involve filtration with .02 micrometre pore filters. 4 The preparation of standards of known concentration using extracts from appropriate untreated wood-based materials can be in accordance with techniques known within the art. 5 According to a further embodiment of the invention accelerated solvent extraction, and soxhlet extraction can also be used. More particularly a preferred embodiment of the invention includes the following steps. 10 a) Primary mechanical fragmentation of the treated wood, wood-based product or engineered wood product. b) Suspension of the wood particles in an appropriate extraction solvent. 15 c) Secondary fragmentation of the suspended wood particles in order to reduce particle size. d) Incubation of the suspended wood particles. 20 e) Separation of the extraction solvent from suspended wood solids. f) Dilution of the extraction solvent into a suitable aqueous sample buffer. 5 g) Determination of the concentration of extracted and diluted preservative using an enzyme-linked immunosorbent assay directed at the preservative of interest. Primary mechanical fragmentation of the wood product (step a above) may include but is 5 not restricted to milling, grinding, shaving, shredding, or sanding or a combination of the above, and may be achieved using equipment such as a Willey mill, or a Silverson as disclosed above. The extraction solvent used in step b above in the preferred form of the inventions is 10 able to both penetrate the wood matrix and solubilise the preservative of interest. Accordingly the solvent may include various organic solvents, water, or aqueous mixtures containing solubility aids such as salts, detergents, or a combination thereof. Secondary mechanical fragmentation of the suspended wood particles (step c above) 15 may include but is not restricted to shearing in an in-line or batch type high shear mixer, treatment with ultrasound, rapid pressurisation and depressurisation as in a French press or a Manton Gaulin mixer, other methods of homogenisation known to those skilled in the art, or a combination of treatments. 20 Incubation of the suspended wood particles (step d above) is in the preferred form of the invention sufficient to allow equilibration of the preservative of interest between the solid and liquid phases of the extraction mixture. It is to be noted that during the incubation the mixture may be stirred, heated, treated with ultrasound, subjected to one 6 or more freeze-thaw cycles, evacuated, pressurised, or a combination of these treatments, in order to facilitate diffusion of the preservative. Separation of the extraction solvent and soluble compounds from the wood solids (step 5 e above) may be achieved by centrifugation, screening, sieving, decantation, or filtration. Preferably the extraction solvent is separated by centrifugation and filtration through a 0.2 micron filter. In order to be quantitated by ELISA technique the extracted preservative is preferably 10 presented in a predominantly aqueous phase. The extraction solvent containing the preservative of interest may be diluted directly into a suitable aqueous sample buffer in the case of a water miscible solvent (step f above), or in the case of a non-water miscible solvent, the solvent may be removed by evaporation and the preservative re dissolved in the sample buffer. 15 Suitable aqueous sample buffers include water or, preferably, a buffered saline solution containing additional compounds as "blocking agents" intended to reduce non-specific binding reactions in the ELISA. Examples include phosphate-buffered saline or tris buffered saline, containing in addition one or more detergents, proteins, or synthetic 20 polymers, in concentrations and combinations known to those skilled in the art. It should be noted that these blocking agents may be included in the aqueous sample buffer or may be added at a later stage during execution of the ELISA. 7 Examples The following example relates to the analyses of commercially manufactured and treated sheets of laminated veneer lumber made of pinus radiata veneers and cured with phenol-formaldehyde resin for the amount of imidacloprid present per one hundred 5 grams of oven dried wood sample. 8 Example 1. Sample Method of extraction and Method of analysis Average Average code cleanup recoveries <40% recoveries >40% w/w w/w ETS 295 Presently accepted method. HPLC with UV detection. X ETS 295 Acidified alcohol with dual HPLC with MS column SPE cleanup. detection. X ETS 295 Ester extraction with GPC HPLC with MS cleanup. detection. X ETS 295 Accelerated solvent HPLC with UV extraction with acidified detection. X alcohol. ETS 285#1 Soxhlet extraction with HPLC with UV ketone. detection. X ETS 285#1 Soxhlet extraction with HPLC with UV haloalkane. detection. X ETS 295 Extraction with acetonitrile GC with MS detection. and derivitised with NBTA. X ETS 295 Extraction with haloalkane GC with MS detection. and derivitised with NBTA. X ETS 295 Extraction with acetonitrile GC with MS detection. and derivitised with HFBA. X ETS 295 Extraction with haloalkane GC with MS detection. and derivitised with HFBA. X ETS 295 Extracted with alcohol enzyme linked immunosorbent assay 9 Thus the present invention provides a method of determining biocides in wood, wood based products and wood engineered products where the method links extraction techniques most commonly employed in GC and HPLC type techniques for analysing 5 compounds in wood, then using ELISA techniques to analyse these extractants. Thus there is a solvent (non-water miscible) extraction technology followed by the water miscible analysis technology. In the past there has been no attempt to employ ELISA form of analysis to wood extractions due to the nature of the solvents used in extraction. The present invention, therefore, employs the more powerful extraction 10 techniques of solvents, but then by evaporating the solvent moving to a water system where the extractant is analysed via a water miscible system. According to the present invention therefore a recovery higher than normal methods of analysis is achieved when immunoassay technology is combined with aspects of 15 chromatographic analyses. Thus, analysis of insecticides in wood and wood-based materials when low dosages of insecticide have been applied can nevertheless yield surprisingly high rates of extraction. Therefore, the present invention provides a method for determining biocides within wood which represents a useful and surprising advance in the art. 20 10

Claims (11)

  1. 2- The method according to claim 1 wherein the pardcle sina of the wood-bised material being analysed Is reduced to between 1000 and 10 micrometres dIamweter.- -
  2. 3. the method according to dalm 1 or 2 wherein the extraction solvent is a Vsubutantialy organic solvent.
  3. 4. The metiad according to any one of claims I to 3 whereln the fitration of extcts Includes filtration with 0.02 micrornetre pore filters.
  4. 5. The method according to any one of daims I to 4 whereld accelerated solvent etcon, and sOxhlat enraction is used, C. Tie RMth aMMMg is am OW d dwis 1 to 5 wMi a dhe pitden~ 4f piid st i by a ~ = nmial neno- a*E CtrlEd ani brow@ Wr mome of WMlng grkintrAouf sh d w ieili C or sang
  5. 7. The method Accoding idan on of dahms I ID 6 Wurhe inUding a secondry mechanical fragment step raded nut in-the presence of orgmnic solvent by one or mre of laring in an hi-line or batch type high shear mxer, umsound. rapid pressudsatlon or depressulstlon or other metids d homogntsadun, prEferably-In-line hich shear mixing. S. The me according to any one of daims 1 to 7 further induding incubaton of suspended wood particles resulting from partide size - reducdon, sufficiem to achieve equilibration of the preservatve of Interest - between solid and lquid phases.
  6. 9. The method according to daimn 8 wherein during incubation the mixture is stirred, heated, treated with ultrasound, subjected to one or more of freeze thaw cydeg, pressurised or a cornbhtation of any of these treatments in order to fadltate diffusion of the preiervatv.
  7. 10. The method awarding to any one of claims 1 to 9 wherein sepaurdon of the extraned solvent and soluble compounds from wood solids is achieved by .centrlfugaton, screening, sierg, decantatipn or filtration. S11. The method according to dalm 10 wherein the extraction solvent Is separated by cennifugatim and filtr n trough a fiter of substntanfy 0.2 'uleron, S 12. The medd acordug to any one of dalms I to 1 Iwherein the 0krce preservat is presented in a pr domanty aeos phase for execon of the presemalve is dMaud div mmiie am aquos m lir
  8. 14. lhM medwd accorn to daim 19wei the assous buer Is a bWHOrM uans solution containing bhiddng ayes.
  9. 15. The metd wardbngto dlaIm .14 wherein the bu-er is phosphatebuffred sine mr td-bul- d saline and One or more detergents, proteins or synthetic pglymers as bloking eeM
  10. 16. The method accordng to caIm 14 or 15 whemrIn the blocking agents are induded in the aqueoMd sample buffer or added at a later ta during execuon of the ELJ5A
  11. 17. The method according to any one of claims 1 to 16 substantially as herein described wth freernce to the Example. *13 9.E4oF CLMMl&
AU2005201412A 2004-04-06 2005-04-05 Method For Determining Biocides Within Wood Active AU2005201412B2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
NZ53215104A NZ532151A (en) 2004-04-06 2004-04-06 A method of quantitation of chemical preservatives within wood using enzyme linked immunosorbent assay technology combined with chromatographic analysis
NZ532151 2004-04-06

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AU2005201412A1 AU2005201412A1 (en) 2005-10-20
AU2005201412B2 true AU2005201412B2 (en) 2010-03-04

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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0592127A1 (en) * 1992-09-28 1994-04-13 Rohm And Haas Company Immunoassay for isothiazolones

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0592127A1 (en) * 1992-09-28 1994-04-13 Rohm And Haas Company Immunoassay for isothiazolones

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NZ532151A (en) 2007-09-28
AU2005201412A1 (en) 2005-10-20

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