High Performance Dioxin/Furan Immunoassay Kit Insert (IN-DF1)
Last Revised: 12/4/98
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E.

Precautions

Important: Please read the following precautions carefully.

Follow precautions and instructions in this insert to achieve the best results.
Safety

*

This kit should only be used by properly trained personnel in an appropriate laboratory environment.

*

Treat PCDD/Fs, solutions that contain PCDD/Fs, and potentially contaminated samples as hazardous materials.
* Use gloves, proper protective clothing, and means to contain and handle hazardous material where appropriate.
* Obtain (if appropriate) permits pertaining to the handling, analysis and transport of dioxin containing materials.
* Stop solution is 1N hydrochloric acid. Handle carefully.
Storage and General Use of Kit
* Do not freeze test kit components or expose them to temperatures greater than 37°C (99°F).
* If desiccant in tube bag is not blue, do not use kit; contact CAPE Technologies.
* Do not expose substrate to direct sunlight.
* Store all test kit components at 2°C to 6°C (36 °F to 43°F) when not in use.
* Storage at ambient temperature (20°C to 27°C or 68°F to 81°F) on the day of use is acceptable. Do not store at ambient temperature for extended periods.
* Allow all reagents to reach ambient temperature (20°C to 27°C or 68°F to 81°F) before beginning the test. This typically requires at least 60 minutes at ambient temperature to warm from recommended storage conditions. Warming will occur faster if bottles are removed from the kit box.
* Do not use test kit components after the expiration date.
* Do not use reagents or test tubes from one test kit with reagents or test tubes from a different test kit.
Sample Preparation
* Because this EIA recognizes many of the congeners in mass labeled internal standard mixtures designed for GC-MS methods, EIA samples must contain either immunoassay internal standards or no internal standards. Consult CAPE Technologies Technical Note TN-001 for specific recommendations regarding mass labeled internal standards.
* Sample extracts must be evaporated to dryness before redissolving in methanol for the EIA. Residual solvents or excessive oil may cause clouding or milkiness upon addition to the sample diluent. If this occurs, the test result will be invalid and the cause should be corrected before repeating the analysis.
EIA Protocol
* When adding standards and samples to the EIA tubes, the methanol solutions must be dispensed directly into the liquid and not above the liquid surface or onto the side of the tube.
Interpretation of Results
* Consult the appropriate CAPE Technologies Application Note for proper interpretation of results.
* Use approved methodologies to confirm any positive results and a portion of the negative results.
* Distribution of PCDD/Fs in samples may vary greatly. The analyst is responsible for determining sampling frequency and distribution.
* Proper quality assurance is the responsibility of the analyst and is strongly encouraged.

 

F.

Materials Provided

*

The contents of this kit are described in a separate Materials List (ML-DF1-60).

G.

Materials Supplied by the User

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Sample preparation supplies and equipment (the SP1 Sample Preparation Kit contains vials and other supplies for rapid preparation of soil or other solid samples; consult the relevant Application Notes or contact CAPE Technologies if additional technical assistance is required).

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HPLC or analytical grade methanol for solvent exchange

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Sample evaporation system for solvent exchange (nitrogen or other gas source)

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Glass tubes or vials for solvent exchange

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 Glass vials with Teflon lined caps for storage of standards after opening ampoules
* 1 glass capillary positive displacement pipettor for dispensing 10 µL samples (Drummond 210 or equivalent)*
* 1 Eppendorf Repeater or Repeater Plus Pipettor and minimum of 6 tips (one 0.5 or 1 mL, four 10 or 12.5 mL, and one 50 mL), for pipetting 50 µL to 1.0 mL volumes (or equivalent repeating pipettor)*
* Artel Differential Photometer or other means for measuring OD of finished immunoassay tubes at 450 nm (alternatives include other tube readers, conventional spectrophotometers, or microplate readers)*
* Marking pen
* Watch or timer
* Reagent grade or bottled distilled water for sample dilution and tube washing
* Basin or other system for capture and disposal of wash water and other waste liquids.

*

See Recommended Equipment List (EL-001) for more information

H.

Suggestions for Pipettor Use
* Please read these suggestions carefully before performing your first EIA.
* Use empty tubes and extra tips to practice your pipettor technique before analyzing samples. For the glass capillary positive displacement pipettor, use methanol; for the Repeater pipettor, use water.
* Use a different tip for each reagent dispensed with the Eppendorf Repeater Plus Pipettor to avoid reagent cross-contamination, especially between conjugate and substrate. Label four 10 or 12.5 mL tips "water", "Conjugate", "Substrate", and "Stop".
* Draw the desired reagent volume into the Repeater pipettor and dispense one portion of reagent back into the container to properly engage the ratchet mechanism. If this is not done, the first volume delivered will be inaccurate.
* To add reagents to the EIA tubes using the Repeater pipettor, pipet down the side of the tube just below the rim to avoid splashback.
* When using the glass capillary positive displacement pipettor for adding samples or standards to EIA tubes, the solution must be dispensed directly into the water in the tube, not above the liquid level or onto the side of the tube.
* The carryover volume of most positive displacement capillaries is minimal, but may affect results if you are going from a high to low PCDD/F concentration. Use a new pipettor tip each time you pipet a new unknown. Rinse or wipe the wire plunger when changing tips to minimize carryover. If capillaries are not changed after each standard, water on the outside of the capillary must be removed by wiping dry to avoid contamination of the standard or sample with water.

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