U.S. Fish & Wildlife Service Emblem
U.S. Fish & Wildlife Service
Analytical Control Facility
A Branch of Ecological Services

Contents:
Who We Are
Services We Provide
Staff & Who to
Contact
Contract
Laboratories
How we assign Labs
Quality Control
Inorganic Lab
Statement of Work
Organic Lab
Statement of Work
Catalog Information
Pricing Guides
Analytical Methods
Contaminant Database

How to Use the Pricing Guides

 
Home
 
Links:
DOI Home Page
USFWS Home Page
DEQ Home Page
Contact us at: chemistry@fws.gov
Privacy & Disclaimer
FOIA
 

Analytical Services, Ltd. (AXYS) Laboratory Methods

Select one of the links below to display the method descriptions associated with AXYS.

Method Code

Method Title

001

Dissection of Tissue Samples

002

Homogenization of Tissue Samples

003

Homogenization of Vegetation

004

Decantation of Water from Sediments

005

Homogenization of Soil and Sediment

006

Homogenization by Stirring

007

Subsampling of Aqueous Samples

008

Homogenization of Blood Samples

011

Soxhlet Extraction of Tissue, Sediment and Soil Samples

012

Solvent Extraction of Water Samples

013

Solvent Extraction of Blood Samples

014

Soxhlet Extraction of XAD Columns

015

Soxhlet Extraction of Particulate Filters

021

Gel Permeation Column Chromatography

022

Chromatographic Cleanup of PCBs, Pesticides and Toxaphene on Florisil

023

Chromatographic Cleanup of Non-ortho-substituted PCBs on Carbon/Celite and Alumina

024

Chromatographic Cleanup of PCB Congeners on Florisil

025

Chromatographic Separation and Cleanup of Dioxins, Furans, Non-ortho-substituted PCBs, Ortho-substituted PCBs, Pesticides and Toxaphene on Carbon/Celite

026

Chromatographic Cleanup of Dioxins, Furans and Non-ortho-substituted PCBs on Alumina

027

Chromatographic Cleanup on Layered Silica

028

Chromatographic Cleanup on PCB Congeners on Florisil, Alumina and Layered Silica

029

Chromatographic Cleanup of PAHs on Layered Silica

030

Chromatographic Cleanup of PCDD/F on Florisil and Alumina Columns

031

HRGC/MS Analysis of PCB Congeners

032

HRGC/MS Analysis of Non-ortho-substituted PCB Congeners

033

HRGC/MS Analysis of Chlorinated Pesticides

034

LRGC/MS Analysis of Chlorinated Pesticides, PCBs (Aroclors), Toxaphene, and PCB Congeners

035

GC/MS/ECNI Analysis of Toxaphene

036

GC/ECD Analysis of Polar Chlorinated Pesticides

037

HRGC/MS Analysis of Dioxins, Furans and Non-ortho-substituted PCBs

038

HRGC/MS/ECNI Analysis of Toxaphene

039

HRGC/MS analysis of PCB Congeners (EPA Method 1668)

040

LRGC/MS Analysis of PAHs

041 HRGC/HRMS Analysis of Polybrominated Diphenylethers (PBDE)

051

Determination of Percent Moisture

052

Determination of Percent Lipid

053

Determination of Grain Size

054

Determination of Total Organic Carbon

055

Determination of Percent Lipid

056 LRGC/MS Analysis of Chlorinated Pesticides, PCBs Congeners, PCB Aroclors, and Toxaphene
057 GC/ECD Analysis of Polar Chlorinated Pesticides
058 HRGC/MS Analysis of Polychlorinated Biphenyl (PCB) Congeners and non-ortho substituted congeners/WHO Congeners
059 HRGC/MS Analysis for Dioxins and Furans
060 HRGC/MS Analysis of Chlorinated Pesticides and Technical Toxaphene
062 HRGC/MS Analysis of Toxaphene Parlars
063 HRGC/MS Analysis of Multi-Residue Pesticides
064 HRGC/MS Analysis of Polychlorinated Naphthalenes
065 HRGC/HRMS Analysis for Polybrominated Diphenylethers (PBDEs)
066 LC-MS/MS Analysis for Hexabromocyclododecane
067 LC-MS/MS Analysis of Tetrabromobisphenol A
068 LRGC/MS Analysis of Parent and Alkylated Aromatic Hydrocarbons
073 LC-MS/MS Analysis of Pharmaceuticals and Personal Care Products
074 LC-MS/MS Analysis of Perfluorinated Organic Compounds (PFCs)
075 LC-MS/MS Analysis for Hormones
085 Homogenization of Tissue Samples

 

 

 

 

 

 

Lab Name: Analytical Services, Ltd.                     Method Code: 001

Dissection of Tissue Samples

The animal is dissected frozen.  Muscle tissue is carefully removed from the bones and skin of the animal with a clean, solvent-rinsed scalpel. Organs are excised from the animal using a solvent rinsed scalpel.  Dissected tissue and/or organ is stored frozen.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 002

Homogenization of Tissue Samples

Tissue samples are thawed then homogenized using clean, solvent rinsed homogenization apparatus suitable to the size of the sample. Small quantities of sample, including organs and bivalves, are homogenized with a Virtis blender.  Muscle tissue is homogenized with an Oster blender.  Whole samples are ground by passing the sample three times through a commercial meat grinder.  Homogenized samples are stored frozen.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 003

Homogenization of Vegetation

Vegetation samples are homogenized frozen.  Grass, leaves, and stalks are cut into small pieces with solvent rinsed scissors and homogenized with a clean Waring Blender.  Large quantities of vegetation are homogenized using a solvent rinsed meat grinder.  Homogenized samples are stored frozen.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 004

Decantation of Water from Sediments

Samples are thawed.  Free standing water is carefully decanted without disturbing the sediment.  If fine particles are decanted with the free water, they are separated by centrifugation and added back to the sample.  Decanted water is discarded.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 005

Homogenization of Soil and Sediment

Samples Soil and sediment samples are thawed.  The sample is transferred to a clean solvent rinsed stainless steel bowl and manually broken up. Rocks larger than 0.5 cm are removed and discarded.  If dry, the sample is passed through a 0.4 cm sieve to remove rocks.  The sample is stirred with a clean spatula until homogeneous.   The homogenized sample is stored frozen.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 006

Homogenization by Stirring

Samples which are received as homogenates are homogenized by stirring with a clean, solvent rinsed spatula just prior to subsampling.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 007

Subsampling of Aqueous Samples

Aqueous samples which need to be subsampled for analysis are homogenized by shaking vigorously and immediately poured into a graduated cylinder.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 008

Homogenization of Blood Samples

Blood samples are thawed and mixed thoroughly by shaking prior to subsampling.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 011

Soxhlet Extraction of Tissue, Sediment and Soil Samples

Animal and plant tissue, soil and sediment samples are extracted by grinding the sample with anhydrous sodium sulphate, spiking with surrogate standards, and refluxing in a soxhlet apparatus for 16 to 20 hours.  The cooled extract is concentrated by rotary evaporation.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 012

Solvent Extraction of Water Samples

Aqueous samples are spiked with surrogate standards and extracted by shaking three times with dichloromethane.  The extract is dried over anhydrous sodium sulphate and concentrated in a Kuderna-Danish concentrator.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 013

Solvent Extraction of Blood Samples

Blood samples, to which surrogate standards have been added are extracted by shaking with an ethanol/ammonium sulphate/hexane mixture, followed by shaking with hexane.  The combined hexane extracts are washed with water and dried over anhydrous sodium sulphate.  The extract is filtered and the solvent removed by rotary evaporation.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 014

Soxhlet Extraction of XAD Columns

XAD resin, from XAD water sampling columns, is dried by filtration through a Millipore filtration apparatus.  The resin is placed in a soxhlet thimble and spiked with surrogate standards.  The sample is soxhlet extracted with dichloromethane.  The extract is concentrated by rotary evaporation.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 015

Soxhlet Extraction of Particulate Filters

Particulate filters from XAD water sampling columns are air-dried.  The filters are placed in a soxhlet thimble and spiked with surrogate standards.  The sample is soxhlet extracted with dichloromethane.  The extract is concentrated by rotary evporation.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 021

Gel Permeation Column Chromatography

Concentrated extracts are loaded onto a calibrated gel permeation column (Biobeads SX-3) to remove high molecular weight interferences.  The column is eluted  with 1:1 dichloromethane and the second fraction collected.  This fraction is concentrated by rotary evaporation, prior to additional chromatographic cleanup procedures.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 022

Chromatographic Cleanup of PCBs, Pesticides and Toxaphene on Florisil

The extract is loaded onto a Florisil column (2.1% deactivated) which is eluted with hexane followed by 15:85 dichloromethane:hexane.  The eluates are collected together (F1).  This fraction contains chlorinated pesticides, toxaphene, and PCB congeners. The fraction is concentrated, an aliquot of recovery standard added and the extract transferred to an autosampler vial in preparation for instrumental analysis.  However, for the analysis of non-ortho-substituted congeners, the fraction is first split, and one half is subject to additional cleanup on carbon/Celite to isolate the non-ortho-substituted PCBs. The Florisil column is then eluted with 1:1 dichloromethane:hexane and the eluate collected (F2).  This fraction contains polar chlorinated pesticides.  The fraction is concentrated, an aliquot of recovery standard added and the extract transferred to an autosampler vial in preparation for instrumental analysis.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 023

Chromatographic Cleanup of Non-ortho-substituted PCBs on Carbon/Celite and Alumina

The first fraction from the Florisil cleanup procedure is eluted through a 4.75% carbon on Celite chromatography column.  The column is eluted with 1:1 cyclohexane:dichloromethane (discard) followed by ethylacetate (discard).  The non-ortho-substituted PCB congeners are eluted with 1:1 toluene:ethylacetate.  The concentrated extract is loaded onto an alumina column (1% deactivated) which is eluted with hexane (discard) followed by 1:1 dichloromethane:hexane (retain).  The eluate is concentrated and an aliquot of recovery standard added in preparation for instrumental analysis.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 024

Chromatographic Cleanup of PCB Congeners on Florisil

The extract is loaded onto a Florisil column (2.1% deactivated) which is eluted with hexane followed by 15:85 dichloromethane:hexane. The eluates are collected together (F1). This fraction contains PCB congeners. The fraction is concentrated, an aliquot of recovery standard added and the extract transferred to an autosampler vial in preparation for instrumental analysis.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 025

Chromatographic Separation and Cleanup of Dioxins, Furans, Non-ortho-substituted PCBs, Ortho-substituted PCBs, Pesticides and Toxaphene on Carbon/Celite

The extract is loaded onto a pre-eluted 4.5% carbon on Celite chromatography column which is eluted with 1:1 cyclohexane:dichloromethane followed by 10:1 ethylacetate:toluene.  The eluates are collected together and concentrated (Fraction E1).  This fraction contains ortho-substituted PCBs, pesticides and toxaphene and requires additional cleanup on Florisil.  The column is inverted, eluted with toluene and the eluate collected (Fraction E2). This fraction contains dioxins, furans and non-ortho-substituted PCBs and requires further cleanup on alumina.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 026

Chromatographic Cleanup of Dioxins, Furans and Non-ortho-substituted PCBs on Alumina

The concentrated extract is loaded onto an alumina column (1% deactivated) which is eluted with hexane.  This fraction is added to the F1/F2 fraction from a Florisil column, if pesticides are part of the analysis.  Otherwise, this fraction is discarded.  The column is then eluted with 1:1 dichloromethane:hexane (retain). The eluate is concentrated and an aliquot of recovery standard added in preparation for instrumental analysis.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 027

Chromatographic Cleanup on Layered Silica

The concentrated extract is loaded onto a layered silica column (layers: neutral, basis, neutral, acidic, acidic).  The column is eluted with hexane.  The eluate is collected and concentrated in preparation for additional chromatographic cleanup procedures.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 028

Chromatographic Cleanup on PCB Congeners on Florisil, Alumina and Layered Silica

The extract is loaded onto a Florisil column (2.1% deactivated) which is eluted with hexane followed by 15:85 dichloromethane:hexane.  The eluates are collected together (F1).  The concentrated fraction is loaded onto an alumina column (1% deactivated) which is eluted with hexane (discard) followed by 1:1 dichloromethane:hexane (retain).  The concentrated fraction is loaded onto a layered silica column (layers: neutral, basic, neutral, acidic, acidic).  The column is eluted with hexane.  The eluate is concentrated and an aliquot of recovery standard added in preparation for instrumental analysis.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 029

Chromatographic Cleanup of PAHs on Layered Silica

The concentrated extract is loaded onto a layered silican column (layers: neutral, basic, neutral, acidic, acidic).  The column is eluted with pentane, and the eluate discarded.  The column is eluted with dichloromethane.  The eluate is collected and concentrated in preparation for additional chromatographic cleanup procedures.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 030

Chromatographic Cleanup of PCDD/F on Florisil and Alumina Columns

The extract is loaded onto a Florisil column (2.1% deactivated) which is eluted with hexane followed by 15:85 dichloromethane:hexane.  The eluates are collected together (F1).  The concentrated fraction is loaded onto an alumina column (1% deactivated) which is eluted with hexane (discard) followed by 1:1 dichloromethane:hexane (retain).  The column is eluted with hexane.  The eluate is concentrated and an aliquot of recovery standard added in preparation for instrumental analysis.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 031

HRGC/MS Analysis of PCB Congeners

High resolution analysis of PCB congeners is carried out using a either a VG 70SE or VG Ultima mass spectrometer.  Each instrument is equipped with a Hewlett Packard 5890 GC, a CTC autosampler and a VAX 3100 data system.  Data are acquired in the Selected Ion Monitoring mode to enhance sensitivity.  The MS is operated at mass resolution 10000.  Chromatographic separation is achieved with a DB-5 capillary column (60 m, 0.25 mm i.d., 0.1 æm film thickness).  A splitless/split injection sequence is used. A calibration solution is run every 12 hours and response factors of PCB congeners are determined. Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 032

HRGC/MS Analysis of Non-ortho-substituted PCB Congeners

High resolution analysis of non-ortho-substituted PCBs is carried out using a VG 70SE mass spectrometer equipped with a Hewlett Packard 5890 GC, a CTC autosampler and a VAX 3100 data system.  Data are acquired in the Selected Ion Monitoring mode to enhance sensitivity.  The MS is operated at mass resolution 10000.  Chromatographic separation is achieved with a DB-5 capillary column (60 m, 0.25 mm i.d., 0.25 æm film thickness).  A splitless/split injection sequence is used. A calibration solution is run every 12 hours and response factors of PCBs are determined.  Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 033

HRGC/MS Analysis of Chlorinated Pesticides

High resolution analysis of pesticides is carried out using a VG 70SE mass spectrometer equipped with a Hewlett Packard 5890 GC, a CTC autosampler and a VAX 3100 data system.  Data are acquired in the Selected Ion Monitoring mode to enhance sensitivity.  The MS is operated at mass resolution 10000.  Chromatographic separation is achieved with a DB-5 capillary column (60 m, 0.25 mm i.d., 0.25 æm film thickness).  A splitless/split injection sequence is used. A calibration solution is run every 12 hours and response factors of pesticides are determined.  Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 034

LRGC/MS Analysis of Chlorinated Pesticides, PCBs (Aroclors), Toxaphene, and PCB Congeners

Analysis of pesticides, PCBs (Aroclors), Total PCBs, toxaphene and individual PCB congeners is carried out using a Finnigan INCOS 50 mass spectrometer equipped with a Varian 3400 GC, a CTC A200S autosampler and a DG10 data system running Incos 50 (Rev 11) software.  The MS is operated at unit mass resolution in the Multiple Ion detection mode.  Chromatographic separation is achieved with a DB-5 capillary column (60 m, 0.25 mm i.d., 0.10 æm film thickness).  A splitless/split injection sequence is used. A calibration solution is run every 12 hours and response factors are determined.  Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 035

GC/MS/ECNI Analysis of Toxaphene

Toxaphene is analyzed using a Finnigan INCOS 50 mass spectrometer equipped with a Varian 3400 GC, a CTC A200S autosampler and a DG10 data system running Incos 50 (Rev 11) software.  The MS is operated in the electron capture negative ionization mode.  Chromatographic separation is achieved with a DB-5 capillary column (60 m, 0.25 mm i.d., 0.10 æm film thickness).  A splitless/split injection sequence is used. A calibration solution is run every 12 hours and response factors are determined.  Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 036

GC/ECD Analysis of Polar Chlorinated Pesticides

The most polar chlorinated pesticides are analyzed using a Hewlett Packard 5890A gas chromatograph equipped with a 63Ni electron capture detector. Chromatographic separation is achieved using a DB-5 capillary column (60 m, 0.25 mm i.d., 0.10 æm film thickness). A splitless/split injection sequence is used. A calibration solution is run every 12 hours and response factors of pesticides are determined.  Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 037

HRGC/MS Analysis of Dioxins, Furans and Non-ortho-substituted PCBs

High resolution analysis of polychlorinated dioxins and furans and non-ortho-substituted PCBs is carried out using a VG Ultima mass spectrometer equipped with a Hewlett Packard 5890 GC, a CTC autosampler and a VAX 3100 data system. Data are acquired in the Selected Ion Monitoring mode to enhance sensitivity.  The MS is operated at mass resolution 10000.  Chromatographic separation is achieved with a DB-5 capillary column (60 m, 0.25 mm i.d., 0.1 æm film thickness).  A splitless/split injection sequence is used. A calibration solution is run every 12 hours and response factors of authentic dioxins, furans and non-ortho-substituted PCBs are determined.  Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 038

HRGC/MS/ECNI Analysis of Toxaphene

Toxaphene is analyzed using a VG Autospec mass spectrometer equipped with a Hewlett Packard 6890 GC, a A200S CTC autosampler and an Alpha data station.  Data are acquired in the Selected Ion Monitoring mode to enhance sensitivity.  The MS is operated at mass resolution 10000 and in the electron capture negative ionization mode.  Chromatographic separation is achieved with a BC-5 capillary column (60 m, 0.25 mm i.d., 0.10 (m film thickness).  A splitless/split injection sequence is used.  A calibration solution is run every 12 hours and response factors are determined.  Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 039

HRGC/MS analysis of PCB Congeners (EPA Method 1668)

High resolution analysis of PCB congeners is carried out using a VG 70SE mass spectrometer equipped with a Hewlett Packard 5890 GC, a CTC autosampler and an Alpha workstation.  Data are acquired in the Selected Ion Monitoring mode to enhance sensitivity.  The MS is operated at mass resolution 10000.  Chromatographic separation is achieved with a SPB-Octyl capillary column (30 m, 0.25 mm i.d., 0.25 um film thickness).  A splitless/split injection sequence is used.  Initial calibration is established using a series of calibration solutions encompassing the working concentration range. Calibration is verified every 12 hours by analysis of a mid-range calibration solution.  Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 040

LRGC/MS Analysis of PAHs

Analysis of PAHs is carried out using a Finnigan INCOS 50 mass spectrometer equipped with a Varian 3400 GC, a CTC A200S autosampler and a Pentium workstation running manufacturer software.  The MS is operated at unit mass resolution in the Multiple Ion detection mode.  Chromatographic separation is achieved on a Restek Rtx-5 chromatography column (30 m, 0.25 mm i.d., 0.25m film thickness), coupled directly to the MS source.  A splitless/split injection sequence is used.  A calibration solution is run every 12 hours and response factors are determined.  Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 041

HRGC/HRMS Analysis of Polybrominated Diphenylethers (PBDE)

Polybrominated diphenylethers are analyzed by high resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS) using a Micromass Ultima or VG70 mass spectrometer (MS) equipped with a Hewlett Packard 5890 or 6890 gas chromatograph, running Micromass software. A DB-5HT capillary chromatography column (30 m, 0.25 mm i.d. x 0.1 um film thickness) is coupled to the MS source. The mass spectrometer is tuned to have a static mass resolution of 5,000. Data are acquired in the Selected Ion Monitoring mode to enhance sensitivity. Initial calibration is established using a series of calibration solutions encompassing the working concentration range. Calibration is verified every 12 hours by analysis of a mid-range calibration solution. Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 051

Determination of Percent Moisture

A weighed subsample is dried for at least 16 hours at 105øC and then reweighed.  Percent moisture is calculated as the percent difference between the dry weight and the wet weight.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 052

Determination of Percent Lipid

Percent lipid determination is carried out on extracts by drying two weighed subsamples of extract at 105øC for 30 minutes.  The extracts are re-weighed.  The percent lipid is calculated as the percent lipid of wet sample weight.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 053

Determination of Grain Size

To determine particle size distribution, samples are pre-treated to remove organic matter, carbonates, soluble salts, and iron oxides.  The percent gravel, sand, silt and clay is determined by a combination of dry sieving, wet sieving and pipetting techniques.

Reference: "Methods of Soil Analysis, Part 1 - Physical and Mineralogical Methods" (Gee and Bauder, 1986) Method 15-4.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 054

Determination of Total Organic Carbon

Total organic carbon is determined by high temperature oxidation of carbon to carbon dioxide which is then measured by means of a nondispersive infrared analyzer.  Total organic carbon is calculated as the percent difference between total and inorganic carbon.

Reference: USEPA Method 9060A

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 055

Determination of Percent Lipid

The concentration of triglycerides, cholestrol and phospholipids are determined enzymatically using Technicon RA-500 Analyser.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 056

LRGC/MS Analysis of Chlorinated Pesticides, PCBs Congeners, PCB Aroclors, and Toxaphene

LRGC/MS Analysis of Chlorinated Pesticides, PCBs Congeners, PCB Aroclors, and Toxaphene

The method utilizes carbon labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Samples are extracted by liquid-liquid extraction (waters) or soxhlet extraction (sediment/soil, tissue, plants). Tissue extracts are first eluted through a gel permeation column to remove lipids. Gel permeation column is an optional cleanup for waters and sediments/soil extracts. All extract are separated into two fractions using Florisil. The final extracts are spiked with an aliquot of recovery standard solution containing 13C12-labelled PCB's prior to instrumental analysis.

The first fraction, E1, is analyzed for low and medium polarity organochlorine pesticides, DDT series, 209 PCB congeners and Technical Toxaphene by gas chromatography with quadrupole (low-resolution) mass spectrometric detection (GC/LRMS). The MS is operated at unit mass resolution in the Multiple Ion detection mode. Chromatographic separation is achieved with a DB-5 capillary. A splitless/split injection sequence is used. A calibration solution is run every 12 hours and response factors are determined. Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

All 209 PCB congeners, with some co-elutions, PCB Homolog groups, Total PCB can be determined. The individual PCB congeners determined can be converted to Aroclor values.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 057

GC/ECD Analysis of Polar Chlorinated Pesticides

The method utilizes carbon labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Samples are extracted by liquid-liquid extraction (waters) or soxhlet extraction (sediment/soil, tissue, plants). Tissue extracts are first eluted through a gel permeation column to remove lipids. Gel permeation column is an optional cleanup for waters and sediments/soil extracts. All extract are separated into two fractions using Florisil. The final extract is spiked with an aliquot of recovery standard solution containing 13C12-labelled PCB's prior to instrumental analysis.

The second fraction, E2, is analyzed for high polarity organochlorine pesticides using gas chromatography with electron capture detection (GC/ECD). Chromatographic separation is achieved using a DB-5 capillary column as the primary column and a DB-17MS capillary column for confirmation. A splitless/split injection sequence is used. A calibration solution is run every 12 hours and response factors of pesticides are determined. Analytes are quantified using the internal standard method of quantification, comparing the area of analyte peak to that of the corresponding surrogate standard and correcting for response factors.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 058

HRGC/MS Analysis of Polychlorinated Biphenyl (PCB) Congeners and non-ortho substituted congeners/WHO Congeners

Two hundred and nine (209) Polychlorinated Biphenyl congeners in a variety of matrices, including tissue, blood/serum/plasma, soil/sediment and water can be measured according to the protocols described in EPA Method 1668A and 1668C and conducted by AXYS MLA-010. Any subset of PCB congeners can be measured including the twelve PCB congeners designated as "toxic' by the World Health Organization (WHO). When all 209 PCBs are measured, Total PCBs, PCB Homologs and PCB Aroclor equivalent concentrations can also be determined. Typical reporting limits range from 0.1 to 0.2 pg/sample.

The method utilizes carbon labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Samples are extracted by liquid-liquid extraction using a separatory funnel or magnetic stirring (waters), soxhlet extraction (sediment/soil, tissue, plants) or by either shaking or solid phase extraction (blood/serum/plasma). Sediments/soils may also be extracted by a soxhlet/Dean-Stark procedure, as an option. The extract is spiked with an aliquot of cleanup surrogate solution containing 13C12-labelled PCBs prior to chromatographic cleanup procedures. Tissue extracts are first eluted through a gel permeation column to remove lipids. Gel permeation column is an optional cleanup for other matrix extracts. The extract is cleaned up on a series of chromatographic columns: layered acid/base silica, Florisil and alumina columns. Sulphur is removed from the extracts by using activated copper. Further cleanup options may be applied in the event of matrix effects. The final extract is spiked with an aliquot of recovery standard solution containing 13C12-labelled PCBs prior to instrumental analysis.

The analysis of sample extracts for PCB congeners is performed by high resolution gas chromatography/high resolution mass spectrometry detection. Two masses from the molecular ion cluster are used to monitor each of the target analytes and 13C12-labelled surrogate standards.

A five point calibration is used to establish linearity. The calibration solutions contain selected analytes of interest covering the working range of the instrument together with the labeled surrogate, cleanup and recovery standards. Alternatively, upon request, a six point, high sensitivity calibration, may be performed. An additional, single point calibration solution containing all target PCB analytes is used to determine the relative response factor and retention times of PCB congeners that are not present in the initial calibration solutions. The single point solution is also used as a calibration verification (CAL-Ver) solution, which is analyzed every 12 hours, to demonstrate stability of calibration.

Target analytes are quantified using the internal standard method of quantification, comparing the area of the quantification ion to that of the 13C-labelled standard and correcting for response factors. The recovery of the surrogate standard is calculated (by internal standard quantification against the recovery standard using an average RRF) and monitored as an indication of overall data quality. Final target concentrations are recovery corrected by this method of quantification.

For the analysis of PCB congeners, sample concentrations are reported using Sample Specific Detection Limit (SDL) as the lower reporting limit.

Aroclor equivalent concentrations may be calculated by converting the summed concentrations of a suite of characteristic PCB congeners to concentrations using empirical factors determined from the analysis of technical Aroclor mixtures.

An optional carbon column cleanup may be required to adequately resolve all toxic PCB congeners from interferences. A 4.5% Carbon/Celite column may be used as part of the initial extract cleanup where it is used just prior to the alumina column. Alternatively, it may be used only after the successful HRGC/HRMS analysis of all 209 PCB congeners has been carried out.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 059

HRGC/MS Analysis for Dioxins and Furans

Tetra- through octa- polychlorinated dibenzodioxins and dibenzofurans in a variety of matrices, including tissue, serum, soil/sediment and water, can be measured according to the protocols described in EPA Method 1613B and conducted by AXYS Method MLA-017. Individual congener and total homologue concentrations as well as the toxicological equivalent (TEQ) for each dioxin and furan congener can be reported. As an option, only 2,3,7,8-TCDD and 2,3,7,8-TCDF maybe reported. Typical reporting limits are 0.5 pg/sample for each congener

The method utilizes carbon labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Samples are extracted by liquid-liquid extraction using either a separatory funnel or magnetic stirring (waters), soxhlet extraction (sediment/soil, tissue, plants) or by liquid-liquid extraction by shaking (blood/serum/plasma). Sediments/soils may also be extracted by a soxhlet/Dean-Stark procedure, as an option. Tissue samples to large (>75 g wet weight) to be extracted by Soxhlet, are extracted by base digestion. The extracts, except blood/serum/plasma extracts, are spiked with an aliquot of cleanup surrogate solution containing 13C4-2,3,7,8-TCDD prior to chromatographic cleanup procedures. The extract is cleaned up on a series of chromatographic columns: layered silver nitrate/acid/base silica (non-tissue extracts) layered acid/base/silica (tissue/blood/serum/plasma extracts), Florisil (tissue/blood/serum/plasma extracts) and alumina/carbon/Celite columns. Further cleanup options may be applied in the event of matrix effects as well as a gel permeation column to remove lipids from tissue extracts. The final extract is spiked with an aliquot of recovery standard solution containing 13C12-labelled TCDDs prior to instrumental analysis.

The analysis of sample extracts for tetra- through octa- polychlorinated dibenzodioxins and dibenzofurans is performed by high resolution gas chromatography/high resolution mass spectrometry detection. Two masses from the molecular ion cluster are used to monitor each of the target analytes and 13C12-labelled surrogate standards. Five additional ions are monitored to check for interference from chlorinated diphenylethers. A second GC column is used for confirmation of 2,3,7,8-TCDF identification.

A five point calibration solutions is used. The calibration solutions contain the analytes of interest covering the working range of the instrument together with the labeled surrogate, cleanup and recovery standards. Alternatively, upon request, a six point, high sensitivity calibration, may be performed. Calibration is verified at least once every 12 hours by analysis of a mid-level calibration solution.

Target analytes are quantified using the internal standard method of quantification, comparing the area of the quantification ion to that of the 13C-labelled standard and correcting for response factors. The recovery of the surrogate standard is calculated (by internal standard quantification against the recovery standard using an average RRF) and monitored as an indication of overall data quality. Final target concentrations are recovery corrected by this method of quantification.

For the analysis of dibenzodioxins and dibenzofurans, sample concentrations are reported using Sample Specific Detection Limit (SDL with a minimum reporting limit of 0.5 pg/sample.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 060

HRGC/MS Analysis of Chlorinated Pesticides and Technical Toxaphene

A suite of 28 organochlorinated pesticide and Technical Toxaphene in a variety of matrices, including tissue, blood/serum/plasma, soil/sediment and water can be measured according to the protocols described in AXYS in house method MLA-028. AXYS MLA-028 has been developed from the best practices in published literature. Typical reporting limits range from 0.01 to 1 ng/sample.

The method utilizes carbon labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Samples are extracted by liquid-liquid extraction using either a separatory funnel or magnetic stirring (waters), soxhlet extraction (sediment/soil, tissue, plants) or by liquid-liquid extraction with shaking (blood/serum/plasma). Tissue extracts are first eluted through a gel permeation column to remove lipids. Gel permeation column is an optional cleanup for other matrix extracts. All extracted are cleaned and separated into two fractions using Florisil. The first fraction, E1, contains non-polar and moderately polar chlorinated pesticides and Toxaphene. Technical Toxaphene (E1) is an optional analysis. The second fraction, E2, contains the more polar chlorinated pesticides. Some extracts may require additional cleanup procedures to remove interference. The final extracts are spiked with an aliquot of the appropriate recovery standard solution containing 13C-labelled pesticides prior to instrumental analysis

The analysis of sample extracts for organochlorine pesticides is performed by high resolution gas chromatography/high resolution mass spectrometry (HRGC/MS) detection. Two masses from the molecular ion cluster are used to monitor each of the target analytes and 13C-labelled surrogate standards. Separate HRCG/MS instruments runs are performed for the E1 and E2 extracts.

A five point calibration is used to establish linearity. The calibration solutions contain suite of analytes of interest covering the working range of the instrument together with the labelled surrogates and recovery standards. Calibration is verified at least once every 12 hours by analysis of a mid-level calibration solution. Separate calibrations are performed for the E1 and the E2 extracts.

Target analytes are quantified using the internal standard method of quantification, comparing the area of the quantification ion to that of the 13C-labelled standard and correcting for response factors. The recovery of the surrogate standard is calculated (by internal standard quantification against the recovery standard using an average RRF) and monitored as an indication of overall data quality. Final target concentrations are recovery corrected by this method of quantification. Technical Toxaphene is quantified by summing the responses of seven peaks and quantifying by internal standard quantification procedures.

For the analysis of organochlorine pesticides and technical toxaphene, sample concentrations are reported using the highest of the Sample Specific Detection Limit (SDL) or the Minimum Reporting Limit. Typical Minimum Reporting Limits in this method are to 2-4 ng/sample for E1 pesticides and 1.6-2 ng/sample for E2 pesticides. Minimum Reporting Limits are prorated based on sample size, final extract volumes, dilutions and splits.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 062

HRGC/MS Analysis of Toxaphene Parlars

The method utilizes a carbon labelled surrogate, 13C12-PCB-159, added at the beginning of the analytical process, prior to extraction, for quantification. Recovery correction of final concentrations in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Samples are extracted by liquid-liquid extraction (waters), soxhlet extraction (sediment/soil, tissue, plants) or by shaking (blood/serum). Tissue extracts are first eluted through a gel permeation column to remove lipids. Gel permeation column may be used as an optional cleanup for water and sediment/ soil extracts. All extracts are separated into two fractions using Florisil. The first fraction, E1, contains non-polar and moderately polar analytes including Toxaphene and Toxaphene congeners/Parlars. The final extract is spiked with an aliquot of recovery standard solution containing 13C12-labelled PCB-153 prior to instrumental analysis.

The extract of the E1 fraction is analyzed by high resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS). The MS is operated at a static (5000) mass resolution in the electron capture negative ionization mode (ECNI) using multiple ion detection (MID) acquiring two characteristic ions for each target analyte and surrogate standard. The linearity of the HRGC/MS is demonstrated using a series of technical Toxaphene solutions that encompass the working concentration range of the instrument. The solutions are prepared from technical Toxaphene, the surrogate standard, and the recovery standard. A calibration standard containing Toxaphene congeners/Parlars, the surrogate and the recovery standard is analyzed every 12 hours, at the beginning and end of a bracket of samples

Target analytes are quantified using the RRFs determined from the Toxaphene congener/Parlars calibration solution. Final concentrations are recovery corrected with respect to the 13C12PCB-159 labelled surrogate.

For the analysis of Toxaphene congeners/Parlars, sample concentrations are reported using Sample Specific Detection Limit (SDL) as the lower reporting limit.

Twenty six (26) Toxaphene congeners/Parlars in a variety of matrices can be measured by this method.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 063

HRGC/MS Analysis of Multi-Residue Pesticides

Organochlorine, organophosphorous, triazine pesticides and pyrethroids in a variety of matrices, including tissue, soil/sediment and water can be measured according to the protocols described in EPA Method 1699 and conducted by AXYS Method MLA-035. Typical reporting limits range from 0.01 to 5 ng/sample.

The method utilizes carbon and deuterium labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Samples are extracted by liquid-liquid extraction using either a separatory funnel or magnetic stirring (waters) or soxhlet extraction (sediment/soil, tissue, plants). Aqueous extracts are cleaned up on an aminopropyl bonded silica solid phase extraction (SPE) column and a micro-silica column. Sediment/soil, tissue and plant extracts are first eluted through a gel permeation column, followed by clean up on an aminopropyl bonded silica solid phase extraction (SPE) column and a micro-silica column. The final extracts are spiked with an aliquot of the recovery standard solution containing a 13C-labelled PCB prior to instrumental analysis

The analysis of sample extracts for multi-residue pesticides is performed by high resolution gas chromatography/high resolution mass spectrometry (HRGC/MS) detection. Two masses from the molecular ion cluster are used to monitor each of the target analytes and carbon and deuterium labelled surrogate standards.

Calibration for this analysis is by single solution bracketing calibration. The calibration solution contains the analytes of interest together with the labelled surrogates and recovery standard. The bracketing calibration solution is analyzed before and after the instrumental analysis of a batch of samples Optionally, a multi-level calibration may be performed using a minimum of 5 calibration solutions that contain the analytes of interest covering the working range of the instrument together with the labelled surrogates and recovery standard. Calibration is verified at least once every 12 hours by analysis of a mid-level calibration solution.

Target analytes are quantified using the internal standard method of quantification, comparing the area of the quantification ion to that of the carbon or deuterium labelled surrogate standard and correcting for response factors. The recovery of the surrogate standard is calculated (by internal standard quantification against the recovery standard using an average RRF) and monitored as an indication of overall data quality. Final target concentrations are recovery corrected by this method of quantification.

For the analysis of multi-residue pesticides, sample concentrations are reported using Sample Specific Detection Limit (SDL) as the lower reporting limit.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 064

HRGC/MS Analysis of Polychlorinated Naphthalenes

Seventy five (75) Polychlorinated Naphthalene congeners (PCNs) in a variety of matrices, including tissue, blood/serum/plasma, soil/sediment and water can be measured according to the protocols described in AXYS in house method MLA-030. AXYS MLA-030 has been developed from the best practices in published literature. Typical reporting limits range from 0.01 to 0.1 pg/sample.

The method utilizes carbon labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Samples are extracted by liquid-liquid extraction using a separatory funnel or magnetic stirring (waters), soxhlet extraction (sediment/soil, tissue, plants) or by liquid-liquid extraction with shaking (blood/serum/plasma). Sediments/soils may also be extracted by a soxhlet/Dean-Stark procedure and tissue may be extracted by base digestion followed by liquid-liquid extraction, as an option. The extract is spiked with an aliquot of cleanup surrogate solution containing prior to chromatographic cleanup procedures. Tissue extracts are first eluted through a gel permeation column to remove lipids. Gel permeation column is an optional cleanup for other matrix extracts. All other extracts are cleaned up on a series of chromatographic columns: layered acid/base silica, carbon/Celite and alumina columns. Further cleanup options such as Florisil or gel permeation columns may be applied in the event of matrix effects. The final extract is spiked with an aliquot of recovery standard solution containing 13C10-labelled PCNs prior to instrumental analysis.

The analysis of sample extracts for PCN congeners is performed by high resolution gas chromatography/high resolution mass spectrometry detection. Two masses from the molecular ion cluster are used to monitor each of the target analytes and 13C10-labelled surrogate standards.

A five point calibration is used to establish linearity. The calibration solutions contain suite of analytes of interest covering the working range of the instrument together with the labeled surrogate, cleanup and recovery standards. Calibration is verified at least once every 12 hours by analysis of a mid-level calibration solution.

Target analytes are quantified using the internal standard method of quantification, comparing the area of the quantification ion to that of the 13C10-labelled standard and correcting for response factors. The recovery of the surrogate standard is calculated (by internal standard quantification against the recovery standard using an average RRF) and monitored as an indication of overall data quality. Final target concentrations are recovery corrected by this method of quantification.

For the analysis of PCN congeners, sample concentrations are reported using Sample Specific Detection Limit (SDL) as the lower reporting limit.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 065

HRGC/HRMS Analysis for Polybrominated Diphenylethers (PBDEs)

Forty six (46) Polybrominated Diphenylethers (PBDEs) in a variety of matrices, including tissue, serum, soil/sediment and water, can be measured according to the protocols described in EPA Method 1614 and conducted by AXYS Method MLA-033. Typical reporting limits for PDBEs range from 1-2 pg/sample for the DiBDE through HpBDE and 10-20 pg/sample for NoBDE and DeBDE.

The method utilizes carbon labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Extraction and cleanup procedures are conducted under low light conditions, eliminating exposure of extracts to direct sunlight and minimizing the exposure to ambient light.

Samples are extracted by liquid-liquid extraction using a separatory funnel or magnetic stirring (waters), soxhlet extraction (sediment/soil, tissue, plants) or by either shaking or solid phase extraction (blood/serum). Sediments/soils may also be extracted by a soxhlet/Dean-Stark procedure, as an option. The extract is spiked with an aliquot of cleanup surrogate solution containing 13C12-labelled BDE prior to chromatographic cleanup procedures. The extract is cleaned up on a series of chromatographic columns: layered acid/base silica, Florisil and alumina columns. Further cleanup options may be applied in the event of matrix effects. The final extract is spiked with an aliquot of recovery standard solution containing 13C12-labelled PBDEs prior to instrumental analysis.

The analysis of sample extracts for PBDEs is performed by high resolution gas chromatography/high resolution mass spectrometry detection. Two masses from the molecular ion cluster are used to monitor each of the target analytes and 13C12-labelled surrogate standards.

A five point calibration solutions is used. The calibration solutions contain selected analytes on interest covering the working range of the instrument together with the labeled surrogate, cleanup and recovery standards. An additional, single point calibration solution containing all target PDBE analytes is used to determine the relative response factor and retention times of PDBE congeners that are not present in the initial calibration solutions. The single point solution is also used as a calibration verification (CAL-Ver) solution, which is analyzed every 12 hours, to demonstrate stability of calibration.

Target analytes are quantified using the internal standard method of quantification, comparing the area of the quantification ion to that of the 13C-labelled standard and correcting for response factors. The recovery of the surrogate standard is calculated (by internal standard quantification against the recovery standard using an average RRF) and monitored as an indication of overall data quality. Final target concentrations are recovery corrected by this method of quantification.

For the analysis of PBDEs, sample concentrations are reported using Sample Specific Detection Limit (SDL) as the lower reporting limit.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 066

LC-MS/MS Analysis for Hexabromocyclododecane

Hexabromocyclododecane, alpha-, beta- and gamma- forms, in solid, aqueous, blood and tissue samples, can be measured according to the protocols in AXYS in house method MLA-070. AXYS MLA-070 has been developed from the best practices in published literature. Typical reporting limits for HBCDD are 2.5 ng/sample for all matrices, exclusive of sample size adjustment.

The method utilizes carbon labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Aqueous samples may be centrifuged prior to analysis depending of solids content within the sample. Samples containing high solids (>1%) may have the particulate and aqueous fractions extracted separately, the extracts can then be cleaned up and analyzed either separately as 2 samples, or the extracts can be combined and cleaned up prior to instrumental analysis. Aqueous samples are liquid-liquid extracted with DCM; solids and tissues are Soxhlet extracted with DCM. Whole blood and serum samples are liquid-liquid extracted with ethanol:hexane:ammonium sulphate. All sample extracts are cleaned up using gel permeation and Florisil chromatography columns. The resulting cleaned up eluates are spiked with recovery standards and analyzed by LC-MS/MS.

The analysis of sample extracts for hormones is performed on a high performance liquid chromatograph reversed phase C18 column coupled to a triple quadrupole mass spectrometer (LC-MS/MS). The mass spectrometer is at unit mass resolution in the Multiple Reaction Monitoring (MRM) mode. Quantification is performed by recording the peak areas of the applicable daughter ion of specified parent ion/daughter transitions.

A series of seven calibration solutions is used to establish initial multi-level calibration. The calibration solutions contain the analytes of interest covering the working range of the instrument together with the labeled surrogate and recovery standards. A mid-level calibration solution is analyzed at least every 12 hours to demonstrate calibration stability.

Target analytes are quantified using the internal standard method of quantification, comparing the area of the quantification ion to that of the 13C-labelled standard and correcting for response factors. The recovery of the surrogate standard is calculated by internal standard quantification against the recovery standard, using an average RRF, and monitored as an indication of overall data quality. Final target concentrations are recovery corrected by this method of quantification.

For the analysis of HBCDD, sample concentrations are reported using the Lower Method Calibration Limit (LMCL) as the lower reporting limit. In cases where the Sample Specific Detection Limit (SDL) is higher than the LMCL, the SDL is used as the lower reporting limit.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 067

LC-MS/MS Analysis of Tetrabromobisphenol A

Tetrabromobisphenol A (TBBPA) analysis in solids, tissues, serum and aqueous samples can be measured according to the protocols in AXYS in house method MLA-079. AXYS MLA-079 has been developed from the best practices in published literature. Typical reporting limits for TBBPA are 10 ng/sample for solid and aqueous samples, and 5 ng/sample for blood, serum and tissue samples.

The method utilizes carbon labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Aqueous samples are filtered, blood are serum samples are sonicated prior to extraction using a solid phase extraction (SPE) cartridge. Solid samples are extracted by shaking; first with dilute acetic acid, then with methanolic ammonium hydroxide solution. Following each different solution solids shaking extraction, the supernatant is collected. Tissue samples are extracted by shaking with methanolic ammonium hydroxide solution. Solid and tissue sample extract cleanup is performed by solid phase extraction (SPE) using anion exchange cartridges. The resulting SPE eluates are spiked with recovery standards and analyzed by LC-MS/MS.

The analysis of sample extracts for TBBPA is performed on a high performance liquid chromatograph reversed phase C18 column coupled to a triple quadrupole mass spectrometer (LC-MS/MS). The mass spectrometer is run at unit mass resolution in the Multiple Reaction Monitoring (MRM) mode. Quantification is performed by recording the peak areas of the applicable daughter ion of specified parent ion/daughter transitions.

A series of six calibration solutions is used to establish initial multi-level calibration. The calibration solutions contain the analytes of interest covering the working range of the instrument together with the labeled surrogate and recovery standards. A mid-level calibration solution is analyzed at least after every 20 samples to demonstrate calibration stability. All calibration solutions for aqueous, solid and serum samples are processed through the same SPE extraction/cleanup procedures as the samples. Tissue samples use a series of solvent based calibration solutions that do not undergo SPE procedures.

Target analytes are quantified using the internal standard method of quantification, comparing the area of the quantification ion to that of the 13C-labelled standard and correcting for response factors. The recovery of the surrogate standard is calculated by internal standard quantification against the recovery standard, using an average RRF, and monitored as an indication of overall data quality. Final target concentrations are recovery corrected by this method of quantification.

For the analysis of hormones, sample concentrations are reported using the Lower Method Calibration Limit (LMCL) as the lower reporting limit. In cases where the Sample Specific Detection Limit (SDL) is higher than the LMCL, the SDL is used as the lower reporting limit.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 068

LRGC/MS Analysis of Parent and Alkylated Aromatic Hydrocarbons

Analysis of the Parent and Alkylated Aromatic Hydrocarbons (PAHs) in aqueous, solid, tissue and whole blood/serum/plasma samples can be measured according to the protocols described in EPA Methods 1625B or 8270C/D and conducted by AXYS method MLA-021. AXYS method MLA-021 is a modified version of EPA 8270C/D, modified by EPA method 1625B for the purposes of inclusion of isotope dilution / recovery correction. PAH analytes are broken down into 3 reporting lists; Lists 1, 2, 3. List 1 contains the Parent PAHs. List 2 contains C1-C4 Naphthalenes and C1-C4 Phenanthrenes/Anthracenes. List 3 contains C1-C4 Fluorenes, C1-C4 Dibenzothiophenes, C1-C4 Fluoranthenes/Pyrenes, C1-C4 Benzo(a)anthraces/Chrysenes and C1-C2 Benzofluoranthenes/Benzopyrenes. Each list also contains a number of other individual alkylated PAHs. Typical reporting limits for List 1-3 PAHs range from 5-10 ng/sample for all matrices.

The method utilizes deuterated labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Aqueous samples may be centrifuged prior to analysis depending of solids content within the sample. Samples containing high solids (>1%) may have the particulate and aqueous fractions extracted separately, the extracts can then be cleaned up and analyzed either separately as 2 samples, or the extracts can be combined and cleaned up prior to instrumental analysis. Aqueous samples are liquid-liquid extracted with dichloromethane (DCM). Solids are Soxhlet extracted with DCM. Optionally, solid samples can undergo base digestion and liquid-liquid extraction with hexane. Tissue samples are base digested and liquid-liquid extracted with hexane. Tissue samples can optionally be Soxhlet extracted with DCM, depending on sample lipid content. Whole blood, serum and plasma samples are liquid-liquid extracted with ethanol:hexane:saturated ammonium sulphate. All sample extracts are cleaned up using silica and gel permeation chromatographic columns and treated with activated copper for removal of excess sulphur. If required, additional extract clean up may be performed using any or all the following methodologies; base wash, alumina chromatographic column, gel permeation chromatographic column. The resulting cleaned up eluates are spiked with recovery standards and analyzed by high-resolution gas chromatography/low resolution mass spectrometry (HRGC/LRMS).

The analysis of sample extracts for PAHs is performed on a high-resolution gas chromatograph (GC) using an RtX-5 nonpolar capillary chromatographic column, coupled to a low resolution mass spectro¬meter (MS) (HRGC/LRMS). The mass spectrometer is run at unit mass resolution in the Multiple Ion Detection (MID) mode. Quantification is performed by recording the peak areas of the applicable daughter ion of specified parent ion/daughter transitions.

A series of at least five calibration solutions is used to establish initial multi-level calibration. The calibration solutions contain the analytes of interest covering the working range of the instrument together with the labeled surrogate and recovery standards. A mid-level calibration solution is analyzed at least once every 12 hours to demonstrate calibration stability. An additional calibration solution is required to be run for the reporting of List 3 analytes. This calibration solution is analyzed at the beginning and end of each batch of samples and is used to establish the relative response factors for the List 3 specific analytes.

Target analytes are quantified using the internal standard method of quantification, comparing the area of the quantification ion to that of the deuterated labelled standard and correcting for response factors. The recovery of the surrogate standard is calculated by internal standard quantification against the recovery standard, using an average RRF, and monitored as an indication of overall data quality. Final target concentrations are recovery corrected by this method of quantification.

For the analysis of PAHs, sample concentrations are reported using Sample Specific Detection Limit (SDL) as the lower reporting limit.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 073

LC-MS/MS Analysis of Pharmaceuticals and Personal Care Products

One hundred and forty one (141) pharmaceuticals and personal care products (PPCP) in aqueous, solid and tissues samples can be measured according to the protocols described in EPA Method 1694 and conducted by AXYS Method MLA-075. AXYS method MLA-075 is a modified version of EPA 1694 for the purposes of inclusion of an extended analyte list. The comprehensive analyte list is broken down into 6 separate analyte lists by extraction and instrumental analysis conditions. Typical reporting limits for PPCPs range from 0.3-60 ng/sample for List 1 and 5 analytes, 6-60 ng/sample for list 2 analytes, 1.5-500 ng/sample for list 3 analytes, 0.3-3.0 ng/sample for list 4 analytes, 0.4-80 ng/sample for list 6 analytes.

The method utilizes carbon and deuterated labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

The presence of free chlorine in aqueous samples may impact the stability of many analytes in MLA-075. If free chlorine is present in the sample, the chlorine is quenched using ascorbic acid.

PPCP analysis requires extraction at two different pH conditions: basic conditions for List 4 analytes; acidic conditions for Lists 1, 2, 3, 5 and 6. All samples are adjusted to the required pH prior to extraction. Solid and tissue samples are extracted by sonication with aqueous buffered acetonitrile and pure acetonitrile and diluted with ultrapure water. Solid and tissue acidic extracts are then treated with EDTA. Solids extracts are filtered and cleaned up by solid phase extraction (SPE) HLB reversed-phase cartridges. Tissue extracts are cleaned up by solid phase extraction (SPE) HLB reversed-phase cartridges and then filtered. All aqueous samples are filtered, EDTA added and cleaned up by solid phase extraction (SPE) HLB reversed-phase cartridges. Mixed phase aqueous/solids samples with significant solids and distinct aqueous and solid phases, such as wastewater influents or process streams, may either be analyzed as an aqueous phase only or as two separate samples, one aqueous and one solid.

The analysis of sample extracts for PPCPs is performed on a high performance liquid chromatograph reversed phase C18 or HILIC column, coupled to a triple quadrupole mass spectrometer (LC-MS/MS). The mass spectrometer is run either run in the positive or negative ionization mode, depending on the PPCP target lists being analyzed, at unit mass resolution in the Multiple Reaction Monitoring (MRM) mode. A total of six instrumental runs are needed to analyze the complete list of analytes, i.e. Lists 1 through 6. Quantification is performed by recording the peak areas of the applicable daughter ion of specified parent ion/daughter transitions.

A series of at least five calibration solutions is used to establish initial multi-level calibration. The calibration solutions contain the analytes of interest covering the working range of the instrument together with the labelled surrogate and recovery standards. A mid-level calibration solution is analyzed every 12 hours or after every 20 samples, whichever occurs first, to demonstrate calibration stability.

Target analytes are quantified using the internal standard method of quantification, comparing the area of the quantification ion to that of the corresponding carbon or deuterated labelled standard and correcting for response factors. The recovery of the surrogate standard is calculated by internal standard quantification against the recovery standard, using an average RRF, and monitored as an indication of overall data quality. Final target concentrations are recovery corrected by this method of quantification.

For the analysis of PPCPs, sample concentrations are reported using the Lower Method Calibration Limit (LMCL) as the lower reporting limit. In cases where the Sample Specific Detection Limit (SDL) is higher than the LMCL, the SDL is used as the lower reporting limit.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 074

LC-MS/MS Analysis of Perfluorinated Organic Compounds (PFCs)

The method utilizes carbon labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Samples are extracted by solid phase extraction (SPE) using weak anion exchange cartridges after filtering (waters), shaking with dilute acetic acid followed by shaking with methanolic ammonium hydroxide (sediment/soil), or shaking with methanolic potassium hydroxide (tissue). For sediment/soil, following extraction, the supernatants are combined, treated with ultra pure carbon powder followed by dilution with water and cleaned up by solid phase extraction (SPE) using weak anion exchange cartridges. For tissue, after centrifugation, an aliquot of the supernatant is diluted with water and cleaned up by solid phase extraction (SPE) using weak anion exchange cartridges. An optional cleanup for tissue supernatants may include ultra pure carbon powder. The resulting eluates are spiked with recovery standards and analyzed by LC-MS/MS.

The analysis of sample extracts for perfluorinated organics is performed on a high performance liquid chromatograph reversed phase C18 column coupled to a triple quadrupole mass spectrometer (LC-MS/MS). The mass spectrometer is run at unit mass resolution in the Multiple Reaction Monitoring (MRM) mode.

A series of at least six calibration solutions is used to establish initial multi-level calibration. The calibration solutions contain the analytes of interest covering the working range of the instrument together with the labeled surrogate and recovery standards. A mid-level calibration solution is analyzed at least after every 20th sample to demonstrate calibration stability. All calibration solutions are processed through the same SPE extraction/cleanup procedures as the samples.

Target analytes are quantified using the internal standard method of quantification, comparing the area of the quantification ion to that of the 13C-labelled standard and correcting for response factors. The recovery of the surrogate standard is calculated (by internal standard quantification against the recovery standard using an average RRF) and monitored as an indication of overall data quality. Final target concentrations are recovery corrected by this method of quantification.

For the analysis of PFCs, sample concentrations are reported using the Lower Method Calibration Limit (LMCL) as the lower reporting limit. In cases where the Sample Specific Detection Limit (SDL) is higher than the LMCL, the SDL is used as the lower reporting limit.

A suite of 9 Perfluorinated Carboxylic Acids (including PFOA), 3 Perfluorinated Sulphonic Acids (including PFOS) and Perfluorooctane Sulfonamide (PFOSA) in a variety of matrices can be measured by this method.

Back to the Top Back to the Top Arrow


Lab Name: Analytical Services, Ltd.                     Method Code: 075

LC-MS/MS Analysis for Hormones

Seventeen (17) hormones in solid, biosolid, and aqueous samples, including influents and effluents, can be measured according to the protocols in AXYS in house method MLA-072. AXYS MLA-072 has been developed from the best practices in published literature. Typical reporting limits for hormones range from 0.8-60 ng/sample for both solid and aqueous matrices.

The method utilizes carbon labelled surrogates, added at the beginning of the analytical process, prior to extraction, for quantification. The use of isotope dilution / recovery correction in each sample allows for compensation of loss of target analyte during the extraction and cleanup process of the analysis.

Aqueous samples are adjusted in pH and filtered and/or centrifuged, to remove solid particulate, prior to being extracted by solid phase extraction (SPE) HLB reversed-phase cartridges. Solid samples are adjusted in pH and extracted by sonication with buffered and pure acetonitrile and diluted with water. The resulting extract is cleaned up using solid phase extraction (SPE) HLB reversed phase cartridges. The aqueous phase, solids phase, or a combined extract from both phases can be selected for analysis. The resulting SPE eluates are spiked with recovery standards and analyzed by LC-MS/MS.

The analysis of sample extracts for hormones is performed on a high performance liquid chromatograph reversed phase C18 column coupled to a triple quadrupole mass spectrometer (LC-MS/MS). The mass spectrometer is run either run in the positive or negative ionization mode, depending on the hormone targets being analyzed, at unit mass resolution in the Multiple Reaction Monitoring (MRM) mode.

A series of at least six calibration solutions is used to establish initial multi-level calibration. The calibration solutions contain the analytes of interest covering the working range of the instrument together with the labeled surrogate and recovery standards. A mid-level calibration solution is analyzed at least after every 20 samples to demonstrate calibration stability. All calibration solutions are processed through the same SPE extraction/cleanup procedures as the samples.

Target analytes are quantified using the internal standard method of quantification, comparing the area of the quantification ion to that of the 13C-labelled standard and correcting for response factors. The recovery of the surrogate standard is calculated by internal standard quantification against the recovery standard, using an average RRF, and monitored as an indication of overall data quality. Final target concentrations are recovery corrected by this method of quantification.

For the analysis of hormones, sample concentrations are reported using the Lower Method Calibration Limit (LMCL) as the lower reporting limit. In cases where the Sample Specific Detection Limit (SDL) is higher than the LMCL, the SDL is used as the lower reporting limit.

Back to the Top Back to the Top Arrow


 

Lab Name: Analytical Services, Ltd.                     Method Code: 085

Homogenization of Tissue Samples

Tissue samples are thawed then homogenized using clean, solvent rinsed homogenization apparatus suitable to the size of the sample. Small quantities of sample, including organs and bivalves, are homogenized with a small blender or by scissors, if quantities are quite small. Larger tissue samples, such as fish Muscle tissue, are is homogenized with a grinder or large blender. Whole samples are ground by passing the sample three times through a commercial meat grinder. Homogenized tissue samples are stored frozen.

Vegetation samples are homogenized frozen. Grass, leaves, and stalks are cut into small pieces with solvent rinsed scissors and homogenized with a clean blender. Large quantities of vegetation are homogenized using a solvent rinsed meat grinder. Homogenized vegetation samples are stored frozen.

Blood/serum samples are thawed and mixed thoroughly by shaking prior to subsampling.

Back to the Top Back to the Top Arrow