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Envirogard™ Toxaphene in Soil Test Kit

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Envirogard™ Toxaphene in Soil Test Kit STRATEGIC DIAGNOSTICS INC. EnviroGard™ Toxaphene in Soil Test Kit 7420000, EPA Method 4040 Intended Use NOTE: Color development is inversely proportional to the Toxaphene concentration. The EnviroGard Toxaphene in Soil Test Kit is a semi- quantitative enzyme immunoassay for the detection of Darker color = lower concentration Toxaphene in soil. The Envirogard Toxaphene in Soil Lighter color = higher concentration Test Kit allows reliable and rapid screening for Toxaphene at 0.5, 2.0 and 10.0 parts per million (ppm) in soil. The determination of the Toxaphene level in an unknown Samples can be screened with a 95% confidence of no sample is interpreted relative to the assay calibrator levels false negatives at the specified action level. using visual comparison or by reading with a spectrophotometer. Test Principles Performance Characteristics The EnviroGard Toxaphene in Soil Test Kit is based on the use of polyclonal antibodies that bind either The EnviroGard Toxaphene in Soil Test Kit will not Toxaphene or Toxaphene-Enzyme Conjugate. These differentiate between Toxaphene and other structurally antibodies are immobilized on the walls of the test tubes. similar compounds, but will detect their presence to When Toxaphene is present in the sample, it competes differing degrees. The following table shows a number of with the Toxaphene-Enzyme Conjugate for a limited compounds and the approximate concentration of each number of Toxaphene binding sites on the immobilized required to yield a positive result at the low calibrator antibodies. (Method Detection Limit or MDL). It also shows the concentration required to inhibit one-half of the color • A sample containing Toxaphene is added to a test developed by the Negative Control (IC50). Concentration tube containing Assay Diluent. Toxaphene-Enzyme is in parts per million (ppm) , or parts per billion (ppb) in Conjugate is then added to the test tube. The soil. Toxaphene-Enzyme Conjugate competes with the Toxaphene for the antibody binding sites. • After the incubation, the unbound molecules are Compound MDL IC50 washed away. Toxaphene 0.5 ppm 2.8 ppm Endrin 3.9 ppb 22 ppb • A clear solution of chromogenic Substrate is then Endosulfan I 6.4 ppb 36 ppb added to the test tube. In the presence of bound Endosulfan II 5.0 ppb 28 ppb Toxaphene-Enzyme Conjugate, the clear Substrate is Dieldrin 7.5 ppb 42 ppb converted to a blue color. One enzyme molecule can convert many Substrate molecules. Heptachlor 6.1 ppb 34 ppb Aldrin 20.7 ppb 116 ppb Since every test tube has the same number of antibody Chlordane 17.9 ppb 100 ppb binding sites and receives the same number of Toxaphene- Gamma-BHC * 0.8 ppm 4.6 ppm Enzyme Conjugate molecules, a sample that contains a Alpha- BHC 3.4 ppm 19 ppm low concentration of Toxaphene allows the antibody to Delta-BHC 7.1 ppm 40 ppm bind many Toxaphene-Enzyme Conjugate molecules. *Gamma-BHC is Lindane Therefore, a low concentration of Toxaphene produces a dark blue solution. Conversely, a high concentration of Toxaphene allows fewer Toxaphene-Enzyme Conjugate molecules to be bound by the antibodies, resulting in a lighter blue solution. 2 EnviroGard Toxaphene in Soil Test Kit Precautions · Do not dilute or adulterate test reagents or use samples not called for in the test procedure; this may · Treat Toxaphene, solutions that contain Toxaphene, give inaccurate results. and potentially contaminated soil samples as hazardous materials. · Tightly recap the Toxaphene calibrator vials to prevent evaporative loss. · Use gloves, proper protective clothing, and methods to contain and handle hazardous material where appropriate. Materials Provided · Store all test kit components at 4°C to 8°C (39°F to · 20 Antibody coated test tubes, 12 X 75 mm 46°F) when not in use. Storage at ambient · 1 vial of Assay Diluent temperature (18°C to 27°C or 64°F to 81°F) on the day of use is acceptable. · 1 vial of Negative Control (Methanol) · Do not freeze test kit components or expose them to · 1 vial of 0.5 ppm Toxaphene Calibrator in methanol (actual concentration is 0.25 ppm) temperatures greater than 37°C (99°F). · 1 vial of 2.0 ppm Toxaphene Calibrator in methanol · Allow all reagents to reach ambient temperature (18°C (actual concentration is 1.0 ppm) to 27°C or 64°F to 81°F) before beginning the test. · 1 vial of 10.0 ppm Toxaphene Calibrator in methanol This typically requires at least 1 hour to warm from (actual concentration is 5.0 ppm) recommended storage conditions. · 1 vial of Toxaphene-Enzyme Conjugate · Do not use test kit components after the expiration · 1 vial of Substrate date. · 1 vial of Stop Solution · Do not use reagents or test tubes from one test kit with reagents or test tubes from a different test kit. · 1 20-Place test tube rack · Use approved methodologies to confirm any positive · 22 Pink (50-250 µL) Gilson Microman® positive results. displacement pipette tips · Soils obtained from areas adjacent to standing water, NOTE: To determine the toxaphene concentration in surface soils collected during or immediately after rain soil, a dilution factor of 2 has been calculated in. or snow, or any soils with relatively high amounts of This factor of 2 is derived from the extraction water (= 30% by weight) should be dried before of the 10 grams of soil with 20 mL of solvent. testing. Contact technical service for recommended methods. Materials Required and Ordered Separately · Distribution of Toxaphene in soils may be highly variable. This variability can be minimized through See “Ordering Information” for the appropriate catalogue numbers. use of a composite sampling technique. Adequate sample number and distribution are the responsibility SDI Sample Extraction Kit of the analyst. Use this kit for the extraction of Toxaphene from soil · Portable spectrophotometer battery must be fully samples. This kit contains enough devices to process 12 samples: charged prior to use. It will not run directly off of AC current. • 12 Extraction jars with screw caps, (each bottle contains 3 stainless steel mixing beads) · Do not expose substrate to direct sunlight. • 12 Filter modules (tops and bottoms) • 12 Ampule crackers • 12 Wooden spatulas EnviroGard Toxaphene in Soil Test Kit 3 • 12 Weigh Canoes Suggestions for Pipettor Use • 12 Disposable Transfer Pipettes · Practice using both pipettors (positive displacement • 12 Ampules containing 20 mL each of 90% Methanol and Repeater pipettor) with water and extra tips Ensys/Envirogard Field Soil Lab (Accessory Kit) before you analyze your samples. Accessory equipment may be rented or purchased from · Use a new tip each time you use the Repeater pipettor Strategic Diagnostics. See “Ordering Information” for the to pipette a different reagent to avoid reagent cross- appropriate catalogue numbers. contamination. Label three 12.5 mL tips "Diluent", "Substrate" and "Stop," and one 5.0 mL tip The accessory kit contains the following items: "Conjugate". Tips can be rinsed thoroughly in clean water and reused. By using the same tip to dispense • Gilson M-25 Microman Positive Displacement the same reagent each time you can avoid cross Pipettor contamination. • EppendorfTM Repeater® Pipettor · Draw the desired reagent volume into the Repeater • Electronic timer pipettor and dispense one portion of the reagent back • Polystyrene test tubes, 12 x 75 mm (for blanking into the container to properly engage the ratchet spectrophotometer) mechanism. If you do not do this, the first volume • Portable balance capable of weighing 10 g delivered may be inaccurate. • Wash bottle · To add reagents using the Repeater pipettor, pipette down the side of the test tube just below the rim. • 5.0 mL Combitips® for the Repeater pipettor -for 0.1 mL to 0.5 mL dispensing volumes (3) · When adding samples and calibrators using the positive displacement pipettor, always pipette below • 12.5 mL Combitips® for the Repeater pipettor the liquid level. Pipet liquid up and down in tip to -for 0.25 mL to 1.250 mL dispensing volumes (6) ensure complete volume transfer. • 50.0 mL Combitip® for the Repeater pipettor (with adapter)-for 1.0 mL to 5.0 mL dispensing volumes (1) · The carryover volume of the positive displacement tips is minimal, but may affect results if you are going • Thirty position foam racks (2) from a high to low Toxaphene concentration. Use a · Artel differential photometer - allows you to measure new pipettor tip each time you pipette a new results in the form of optical density values. These unknown. values can be used for objective record keeping and quality assurance. It is included in the Assay Procedure Ensys/Envirogard Field Soil Lab. Collect/Store the Sample ® NOTE: Order replacement Combitips and positive displacement tips separately. See the “Ordering The following steps explain how to properly collect and Information" section. store your samples. Materials Required but Not Provided 1. Collect soil in appropriately sized and labeled · Protective clothing (e.g., latex gloves) containers. · Absorbent paper for blotting test tubes 2. Take care to remove excess twigs, organic matter, · Liquid and solid waste containers and rocks or pebbles from the soil sample to be tested. · Tap or distilled water for test tube washes 3. Soils obtained from areas adjacent to standing water, · Marking pen surface soils collected during or immediately after rain or snow, or any soils with relatively high · Calculator (optional) amounts of water (= 30% by weight) should be 4 EnviroGard Toxaphene in Soil Test Kit dried before testing. Contact Technical Services for 5. Use the positive displacement pipettor to add the recommended methods. Negative Control (methanol), the Toxaphene Calibrators, and the Sample extracts to the 4. Store soil samples at 4°C (39°F). appropriate test tubes. Use a clean pipette tip for each addition.
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