The Hydrocodone Enzyme Immunoassay is an in vitro diagnostic test intended for the qualitative and semi‑quantitative determination of hydrocodone in human urine. The cutoffs for the assay are 100 ng/mL and 300 ng/mL when calibrated against hydrocodone. The assay is designed for prescription use on the Roche cobas c analyzers.

The semi‑quantitative mode is for purposes of (1) enabling laboratories to determine an appropriate dilution of the specimen for verification by a confirmatory method such as gas or liquid chromatography/mass spectrometry (GC‑MS or LC‑MS) or (2) permitting laboratories to establish quality control procedures.

The assay provides only a preliminary analytical result. A more specific alternative analytical chemistry method must be used in order to obtain a confirmed analytical result. Gas or Liquid Chromatography/Mass Spectrometry (GC‑MS or LC‑MS) are the preferred confirmatory methods.

The Hydrocodone Enzyme Immunoassay is a homogeneous enzyme immunoassay ready‑for‑use liquid reagent. The assay is based on competition between drug in the sample and drug labeled with the enzyme glucose‑6‑phosphate dehydrogenase (G6PDH) for a fixed amount of antibody in the reagent.

1. A preliminary positive result from the assay indicates only the presence of hydrocodone.

2. The test is not intended for quantifying this single analyte in patient samples.

3. A preliminary positive result does not necessarily indicate drug abuse.

4. A negative result does not necessarily mean a person did not take illegal drugs.

5. There is a possibility that other substances and/or factors not listed above may interfere with the test and cause incorrect results (e.g., technical or procedural error, fluid intake, endogenous or exogenous interferents).

6. Preliminary positive results should be confirmed by other affirmative, analytical chemistry methods (e.g., chromatography), preferably GC‑MS or LC‑MS.

7. The test is designed for use with human urine only.

8. The test is not for therapeutic drug monitoring.

ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on cobas c systems. All special wash programming necessary for avoiding carry-over is available via the cobas link. The latest version of the carry-over evasion list can be found with the NaOHD/SMS/SCCS Method Sheet. For further instructions, refer to the operator’s manual.

HYD1Q: ACN 20830: for qualitative assay, 100 ng/mL

HYD1S: ACN 20832: for semi‑quantitative assay, 100 ng/mL

HYD3Q: ACN 20831: for qualitative assay, 300 ng/mL

HYD3S: ACN 20833: for semi‑quantitative assay, 300 ng/mL

Ready for use

Filling the cobas c pack:

The HYD cobas c pack is now ready for use.

NOTE: Solutions must be at the reagent compartment storage temperature of the analyzer before performing assays.

Note
Always use a new cobas c pack when preparing fresh reagent. Never reuse accessories designed for single use, as this may result in reagent contamination and could affect test results. If the cobas c pack bottles are not filled correctly, this may result in faulty reagent pipetting and could cause erroneous results.

For further information about the assay test definitions refer to the application parameters setting screen of the corresponding analyzer and assay.

For the cutoff calibrator a value of “0” is encoded in the e‑barcode in order to ensure flagging of positive samples with >Test and negative absorbance values for negative samples.

Calibration interval may be extended based on acceptable verification of calibration by the laboratory.

Traceability: This method has been standardized against a primary reference method (GC‑MS or LC‑MS).

Representative performance data on a Beckman Coulter AU480 analyzer are given below. Data was validated on a Roche cobas c 501, cobas c 502, and cobas c 503 analyzer and did not show any significant differences between the analyzers. Results obtained in individual laboratories may differ.

Hydrocodone (100 ng/mL Cutoff)

The assay range from 0 ng/mL to 300 ng/mL was tested in qualitative (∆OD, mAU) and semi‑quantitative (ng/mL) mode using a modified NCCLS protocol. Results shown below were obtained by testing all samples in replicates of 2, 2 runs per day for 22 days on a Beckman Coulter AU480 automated clinical chemistry analyzer.

Qualitative analysis: Typical results (measured by ∆OD, mAU) are as follows:

Semi‑quantitative analysis: The following concentrations were determined with reference curves from 5 calibrators. Typical results were measured in ng/mL. Preliminary positive/negative results are as follows:

Hydrocodone (300 ng/mL Cutoff)

The assay range from 0 ng/mL to 600 ng/mL was tested in qualitative (∆OD, mAU) and semi‑quantitative (ng/mL) mode using a modified NCCLS protocol. Results shown below were obtained by testing all samples in replicates of 2, 2 runs per day for 22 days on a Beckman Coulter AU480 automated clinical chemistry analyzer.

Qualitative analysis: Typical results (measured by ∆OD, mAU) are as follows:

Semi‑quantitative analysis: The following concentrations were determined with reference curves from 5 calibrators. Typical results were measured in ng/mL. Preliminary positive/negative results are as follows:

Hydrocodone is an opioid compound derived from codeine. Hydrocodone acts on μ‑opioid receptors.

The hydrocodone metabolite, hydromorphone, is also a minor urinary metabolite of morphine. Initial studies suggest that hydromorphone may have advantages as an analgesic as compared to morphine and is 7‑10 times more potent than morphine.

For timing of reagent pipetting refer to the test definition section.

For USA: Caution: Federal law restricts this device to sale by or on the order of a physician.

Good laboratory practices recommend the use of at least two levels of control specimens (one positive and one negative control near the cutoff) to ensure proper assay performance. Controls should be run with each new calibration and after specific maintenance or troubleshooting procedures as detailed in the instrument manual. Each laboratory should establish its own control frequency. If any trends or sudden change in control value are observed, review all operating parameters, or contact Lin-Zhi International, Inc. (LZI) technical support for further assistance. Laboratories should comply with all federal, state, and local laws, as well as all guidelines and regulations.

For quality control, use control materials as listed in the \"Order information\" section.

In addition, other suitable control material can be used.

The control intervals and limits should be adapted to each laboratory’s individual requirements. It is recommended to perform quality control always after lot calibration and subsequently at least every 14 days.

Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.

Follow the applicable government regulations and local guidelines for quality control.

Urine samples may be collected in plastic or glass containers. Some plastics may adsorb drugs. Use of plastics such as polyethylene is recommended.

See the limitations and interferences section for details about possible sample interferences.

Sample stability claims were established by experimental data by the manufacturer or based on reference literature and only for the temperatures/time frames as stated in the method sheet. It is the responsibility of the individual laboratory to use all available references and/or its own studies to determine specific stability criteria for its laboratory.

Handle all urine specimens as if they were potentially infectious.

The Hydrocodone Enzyme Immunoassay is intended for the qualitative and semi‑quantitative determination of hydrocodone in human urine at a cutoff value of 100 ng/mL and 300 ng/mL. The assay is designed for prescription use with the Roche cobas c 501/502 automated clinical chemistry analyzers.

The assay provides only a preliminary analytical result. A more specific alternative analytical chemistry method must be used in order to obtain a confirmed analytical result. Gas or Liquid Chromatography/Mass Spectrometry (GC‑MS or LC‑MS) are the preferred confirmatory methods.

The hydrocodone assay is a homogeneous enzyme immunoassay ready‑for‑use liquid reagent. The assay is based on competition between drug in the sample and drug labeled with the enzyme glucose‑6‑phosphate dehydrogenase (G6PDH) for a fixed amount of antibody in the reagent.

1. A preliminary positive result from the assay indicates only the presence of hydrocodone.

2. The test is not intended for quantifying this single analyte in patient samples.

3. A preliminary positive result does not necessarily indicate drug abuse.

4. A negative result does not necessarily mean a person did not take illegal drugs.

5. There is a possibility that other substances and/or factors not listed above may interfere with the test and cause incorrect results (e.g., technical or procedural error, fluid intake, endogenous or exogenous interferents).

6. Preliminary positive results should be confirmed by other affirmative, analytical chemistry methods (e.g., chromatography), preferably GC‑MS or LC‑MS.

7. The test is designed for use with human urine only.

8. The test is not for therapeutic drug monitoring.

Materials required (but not provided):

For cobas c 501 analyzer:

HYD1Q: ACN 184: for qualitative assay, 100 ng/mL

HYD1S: ACN 193: for semi‑quantitative assay, 100 ng/mL

HYD3Q: ACN 190: for qualitative assay, 300 ng/mL

HYD3S: ACN 196: for semi‑quantitative assay, 300 ng/mL

For cobas c 502 analyzer:

HYD1Q: ACN 8184: for qualitative assay, 100 ng/mL

HYD1S: ACN 8193: for semi‑quantitative assay, 100 ng/mL

HYD3Q: ACN 8190: for qualitative assay, 300 ng/mL

HYD3S: ACN 8196: for semi‑quantitative assay, 300 ng/mL

Ready for use

Filling the cobas c pack:

The HYD cobas c pack is now ready for use.

NOTE: Solutions must be at the reagent compartment storage temperature of the analyzer before performing assays.

Note
Always use a new cobas c pack when preparing fresh reagent. Never reuse accessories designed for single use, as this may result in reagent contamination and could affect test results. If the cobas c pack bottles are not filled correctly, this may result in faulty reagent pipetting and could cause erroneous results.

Deselect Automatic Rerun for these applications in the Utility menu, Application screen, Range tab.

^{b) See Results section}

Calibration interval may be extended based on acceptable verification of calibration by the laboratory.

Traceability: This method has been standardized against a primary reference method (GC‑MS or LC‑MS).

Representative performance data on a Beckman Coulter AU480 analyzer are given below. Data was validated on a Roche cobas c 501 and cobas c 502 analyzer and did not show any differences between the analyzers. Results obtained in individual laboratories may differ.

Hydrocodone (100 ng/mL Cutoff)

The assay range from 0 ng/mL to 300 ng/mL was tested in qualitative (∆OD, mAU) and semi‑quantitative (ng/mL) mode using a modified NCCLS protocol. Results shown below were obtained by testing all samples in replicates of 2, 2 runs per day for 22 days on a Beckman Coulter AU480 automated clinical chemistry analyzer.

Qualitative analysis: Typical results (measured by ∆OD, mAU) are as follows:

Semi‑Quantitative analysis: The following concentrations were determined with reference curves from 5 calibrators. Typical results were measured in ng/mL. Preliminary positive/negative results are as follows:

Hydrocodone (300 ng/mL Cutoff)

The assay range from 0 ng/mL to 600 ng/mL was tested in qualitative (∆OD, mAU) and semi‑quantitative (ng/mL) mode using a modified NCCLS protocol. Results shown below were obtained by testing all samples in replicates of 2, 2 runs per day for 22 days on a Beckman Coulter AU480 automated clinical chemistry analyzer.

Qualitative analysis: Typical results (measured by ∆OD, mAU) are as follows:

Semi‑Quantitative analysis: The following concentrations were determined with reference curves from 5 calibrators. Typical results were measured in ng/mL. Preliminary positive/negative results are as follows:

Hydrocodone is an opioid compound derived from codeine. Hydrocodone acts on μ‑opioid receptors.

The hydrocodone metabolite, hydromorphone, is also a minor urinary metabolite of morphine. Initial studies suggest that hydromorphone may have advantages as an analgesic as compared to morphine and is 7‑10 times more potent than morphine.

For USA: Caution: Federal law restricts this device to sale by or on the order of a physician.

Good laboratory practices recommend the use of at least two levels of control specimens (one positive and one negative control near the cutoff) to ensure proper assay performance. Controls should be run with each new calibration and after specific maintenance or troubleshooting procedures as detailed in the instrument manual. Each laboratory should establish its own control frequency. If any trends or sudden change in control value are observed, review all operating parameters, or contact Lin‑Zhi International, Inc. (LZI) technical support for further assistance. Laboratories should comply with all federal, state, and local laws, as well as all guidelines and regulations.

Urine samples may be collected in plastic or glass containers. Some plastics may adsorb drugs. Use of plastics such as polyethylene is recommended.

See the limitations and interferences section for details about possible sample interferences.

Sample stability claims were established by experimental data by the manufacturer or based on reference literature and only for the temperatures/time frames as stated in the method sheet. It is the responsibility of the individual laboratory to use all available references and/or its own studies to determine specific stability criteria for its laboratory.

Handle all urine specimens as if they were potentially infectious.