2 Summary
Phencyclidine, also known as PCP and “angel dust,” is a synthetic drug that was orginally developed for its anesthetic properties but is now a drug of abuse used solely for its potent hallucinogenic effects. It may be self-administered in a variety of ways, including ingestion, inhalation, and intravenous injection. Phencyclidine is absorbed well and quickly, and concentrates in the brain and fatty tissues.2 Excretion patterns vary widely, ranging from several hours to a couple of weeks. Phencyclidine is excreted in the urine unchanged, as conjugated Phencyclidine Assay metabolites, and primarily as unidentified compounds. The Emit® II Plus Phencyclidine Assay tests for phencyclidine in human urine. It also detects August 2010 9J052.4D_C the analog 1-[1-(2-thienyl)-cyclohexyl]piperidine (TCP). High concentrations of several phencyclidine metabolites and analogs can also produce positive results in the assay. Positive results for specimens containing other compounds structurally unrelated to phencyclidine usually have not been observed. Methods historically used for detecting phencyclidine in biological fluids include thin-layer chromatography,3 gas chromatography,4 ultraviolet spectroscopy, enzyme immunoassay, and radioimmunoassay.5, 6 While confirmation techniques other than GC/MS may be adequate for some drugs of abuse, GC/MS is generally accepted as a vigorous confirmation technique for all drugs, since it provides the best level of confidence in the result.1 Catalog Quantity/ Number Product Description Volume 3 Methodology OSR9J229 Emit® II Plus Phencyclidine Assay The Emit® II Plus Phencyclidine Assay is a homogeneous enzyme immunoassay technique OSR9J618 R1 (Antibody/Substrate Reagent 1) 2 x 27 mL used for the analysis of specific compounds in human urine.6 The assay is based on competition between drug in the specimen and drug labeled with the enzyme glucose-6-phosphate OSR9J648 R2 (Enzyme Reagent 2) 2 x 13 mL dehydrogenase (G6PDH) for antibody binding sites. Enzyme activity decreases upon binding to 9A509UL Emit® Calibrator/Control Level 0* 1 x 14 mL the antibody, so the drug concentration in the specimen can be measured in terms of enzyme 9A549UL Emit® Calibrator/Control Level 2 (12.5)* 1 x 14 mL activity. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH, resulting 9A569UL Emit® Calibrator/Control Level 3 (25)* 1 x 14 mL in an absorbance change that is measured spectrophotometrically. Endogenous serum G6PDH 9A589UL Emit® Calibrator/Control Level 4 (75)* 1 x 14 mL does not interfere because the coenzyme NAD functions only with the bacterial (Leuconostoc 9A609UL Emit® Calibrator/Control Level 5 (100)* 1 x 14 mL mesenteroides) enzyme employed in the assay. *Required for calibrating the Emit® II Plus Phencyclidine Assay. Sold separately. To determine the appropriate calibrators required for use, see Table 1. 4 Reagents Note: Reagents and calibrators/controls are shipped ready to use in liquid form. No reconstitution is required. Reagents contain the following substances: Note: Reagents 1 and 2 are provided as a matched set. They should not be interchanged with Sheep polyclonal antibodies to phencyclidine (0.2 µg/mL), glucose-6-phosphate (5.5 mM), components of kits with different lot numbers. nicotinamide adenine dinucleotide (3.5 mM), bovine serum albumin, phencyclidine labeled Note: These reagents are qualified for use with these calibrators only. However, other with G6PDH (0.47 U/mL), Tris buffer, preservatives, and stabilizers. material may be used for quality control purposes. Precautions Table 1 — Emit® Calibrators/Controls for Use in Qualitative or Semiquantitative Analysis • For in vitro diagnostic use. Qualitative Analysis Semiquantitative Analysis • Reagent 1 contains nonsterile sheep antibodies. Required Concentration Required Concentration • Reagent 2 contains nonsterile mouse antibodies. Cal/Control of Phencyclidine Cal/Control of Phencyclidine • Reagents 1 and 2 contain nonsterile bovine serum albumin. Level (ng/mL) Level (ng/mL) • Do not use after expiration date. Level 0 0 Level 0 0 Level 3 25 Level 2 12.5 • Turbid or yellow reagents may indicate contamination or degradation and must be discarded. Level 5 100 Level 3 25 Level 4 75 Preparation of Reagents Level 5 100 The Emit® II Plus Phencyclidine Assay reagents are provided ready to use; no preparation is necessary. Note: The Emit® Calibrators/Controls contain the stated concentrations of phencyclidine listed in Table 1. Emit® Calibrator/Control Levels 2, 3, 4, and 5 contain additional drugs of Storage of Assay Components abuse that do not affect the assay. A calibrator/control is used either as a calibrator or as a • Improper storage of reagents can affect assay performance. control when the assay is used for qualitative analysis. When a calibrator/control is used as a calibrator, the other level calibrators/controls (above or below it, as listed above) are used as • When not in use, store reagents upright at 2–8°C and with screw caps tightly closed. controls. See the Emit® Calibrators/Controls instructions for use. • Unopened reagents are stable until the expiration date printed on the label if stored upright at 2–8°C. 1 Intended Use • Do not freeze reagents or expose them to temperatures above 32°C.
The Emit® II Plus Phencyclidine Assay is a homogeneous enzyme immunoassay with a 25 ng/mL cutoff (SAMHSA initial test cutoff level). The assay is intended for use in the qualitative and 5 Specimen Collection and Preparation semiquantitative analyses of phencyclidine in human urine. These reagents are packaged specifically for use on a variety of AU® Clinical Chemistry Systems. • Urine specimens may be collected in plastic (ie, polypropylene, polycarbonate, polyethylene) or glass containers. Some plastics can adsorb certain drugs. The Emit® II Plus Phencyclidine Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used to obtain a confirmed analytical • If not analyzed immediately, specimens may be stored unrefrigerated for up to 7 days result. Gas chromatography/mass spectrometry (GC/MS) is the preferred confirmatory following collection. After 7 days, specimens should be stored frozen. 1 method. Other chemical confirmation methods are available. Clinical consideration and • Frozen specimens must be thawed and mixed thoroughly prior to analysis. professional judgment should be applied to any drug-of-abuse test result, particularly when preliminary positive results are used. • Specimens with high turbidity should be centrifuged before analysis. • The recommended pH range for urine specimens is 3.0–10.0. • Adulteration of the urine specimen may cause erroneous results. If adulteration is suspected, obtain another specimen. • Human urine specimens should be handled and treated as if they are potentially infectious. 6 Procedure Table 2 — Within-Run and Total Precision for the Emit® II Plus Phencyclidine Assay Within-Run Precision Total Precision Materials Provided Phencyclidine Emit® II Plus Phencyclidine Assay 25 ng/mL Cutoff Control Control Cutoff Control Control Reagent 1 Cutoff Cal 75% 125% Cal 75% 125% Reagent 2 Mean 441 417 471 441 417 471 Materials Required But Not Provided (mAU/min) Emit® Calibrators/Controls SD 3.9 2.9 4.0 5.3 4.2 5.7 Commercially available controls (see Quality Control, Semiquantitative Analysis) %CV 0.9 0.7 0.8 1.2 1.0 1.2 Refer to the instrument User’s Guide for appropriate instrument checks and maintenance instructions. Comparative Analysis Calibration Clinical urine specimens were analyzed using Emit® II Plus Phencyclidine Assay on the Qualitative Analysis AU400/AU600 Clinical Chemistry System and using Emit® II Phencyclidine Assay on the Run the Emit® Calibrator/Control Level 3 (25 ng/mL Cutoff) in duplicate. Validate the calibration SYVA®-30R Biochemical System. Specimens positive by either method contained phencyclidine by running controls (see Quality Control). Refer to the analyzer User’s Guide or the Application ranging from 14.9 ng/mL to 78.5 ng/mL. Table 3 summarizes the number of positive/negative Sheet for instrument settings. Recalibrate as indicated by control results. results identified and the percent agreement with the SYVA®-30R Biochemical System. Semiquantitative Analysis Prepare a calibration curve by running a reagent blank (blue rack) and the Table 3 — Summary of Comparative Analysis Emit® Calibrators/Controls Level 2 (12.5 ng/mL), Level 3 (25 ng/mL), Level 4 (75 ng/mL), and Assay Positive Negative % Agreement Level 5 (100 ng/mL). Validate the calibration by running controls (see Quality Control). Refer to the instrument User’s Guide or the Application Sheet for instrument settings. Recalibrate as Phencyclidine 49 54 99 indicated by control results. Analytical Recovery Quality Control Negative human urine specimens were spiked with known concentrations of phencyclidine. Qualitative Analysis Specimens spiked with drug concentrations lower than the cutoff concentration and tested Validate the calibration by assaying controls. Ensure that the result from the qualitatively were correctly identified as negative 100% of the time. Specimens spiked with drug Emit® Calibrator/Control (Level 0 [0 ng/mL] or Level 5 [100 ng/mL]) relates appropriately to concentrations greater than the cutoff were correctly identified as positive 100% of the time. the cutoff calibrator (Emit® Calibrator/Control Level 3 [25 ng/mL]) result. Once calibration is Table 4 summarizes the results on semiquantitative analysis of the specimens. validated, run urine specimens. Semiquantitative Analysis Table 4 — Semiquantitative Analysis of Phencyclidine-Spiked Samples Validate the calibration curve by assaying commercial controls. Ensure that control results fall Concentration (ng/mL) Mean (ng/mL) within acceptable limits as defined by your laboratory. Once the calibration curve is validated, run urine specimens. 12.5 13.0 17.5 16.6 Evaluation and Interpretation of Results 40 45.5 When the Emit® II Plus Phencyclidine Assay is used as a qualitative assay, the amount of phencyclidine or phencyclidine metabolites or analogs detected by the assay in any given 50 58.5 specimen cannot be estimated. The assay results distinguish between positive and negative 90 86.2 specimens—positive indicating specimens contain phencyclidine or phencyclidine metabolites or analogs. Negative results may indicate specimens do not contain phencyclidine or phencyclidine metabolites or analogs, or phencyclidines are present in concentrations below the cutoff level Specificity for this assay. The Emit® II Plus Phencyclidine Assay detects phencyclidine or phencyclidine metabolites or • A specimen that gives a change in rate value equal to or higher than the rate of the cutoff analogs in human urine. calibrator level is interpreted as positive. Tables 5 and 6 list the compounds this assay detects and the levels at which the compounds • A specimen that gives a change in rate value lower than the rate of the cutoff calibrator level is have been found to give a response approximately equivalent to that of the cutoff calibrator interpreted as negative. (Emit® Calibrator/Control Level 3, 25 ng/mL). Each concentration represents the reactivity level for the stated compound when it is added to a negative urine specimen. These concentrations When used semiquantitatively, the Emit® II Plus Phencyclidine Assay yields approximate, are within the range of the levels found in urine following use of the compound or, in the case of cumulative concentrations of the drugs detected by the assay (See Section 8, Specific metabolites, the parent compound. If a specimen contains more than one compound detected by Performance Characteristics, Analytical Recovery or Specificity). the assay, lower concentrations than those listed in Tables 5 and 6 may combine to produce a rate approximately equivalent to or greater than that of the cutoff calibrator. 7 Limitations of the procedure Table 5 — Concentrations of Phencyclidine Compounds That Produce a Result Approximately Equivalent to the 25 ng/mL Phencyclidine Cutoff • The assay is designed for use only with human urine. Compound Concentration (ng/mL) • A positive result from the assay indicates only the presence of phencyclidine but does not indicate or measure intoxication. N,N-Diethyl-1-phenylcyclohexylamine (PCDE) 234 • Boric acid is not recommended as a preservative for urine. 1-(4-Hydroxypiperidino)phenylcyclohexane 420 • Other substances and/or factors not listed (eg, technical or procedural errors) may interfere with the test and cause false results. 1-(1-Phenylcyclohexyl)morpholine (PCM) 41 • Interpretation of results must take into account that concentrations of phencyclidine or 1-(1-Phenylcyclohexyl)pyrrolidine (PCPy) 54 phencyclidine metabolites or analogs in urine can vary extensively with fluid intake and other 4-Phenyl-4-piperidinocyclohexanol 32 biological variables. • Immunoassays that produce a single result in the presence of a drug and its metabolites 1-[1-(2-Thienyl)-cyclohexyl]morpholine (TCM) 80 cannot fully quantitate the concentration of individual components. 1-[1-(2-Thienyl)-cyclohexyl]piperidine (TCP) 37 1-[1-(2-Thienyl)-cyclohexyl]pyrrolidine (TCPy) 83 8 Specific Performance Characteristics Table 6 — Concentrations (µg/mL) of Compounds That Produce a Result Approximately The information presented in this section is based on Emit® II Plus Phencyclidine Assay studies Equivalent to the 25 ng/mL Phencyclidine Cutoff performed on the AU400®/AU600® Clinical Chemistry System. Positive specimens were confirmed by GC/MS. Refer to the Application Sheets for other AU Clinical Chemistry Systems Compound Concentration (µg/mL) and for additional information. Results may vary due to analyzer-to-analyzer differences. The following performance characteristics represent total system performance and should not be Dextromethorphan 120 interpreted to refer only to reagents. Dextrorphan 97 Precision Meperidine 67 Within-run precision was determined by assaying two replicates of each cutoff calibrator/control and positive and negative controls twice a day for 20 days (N=80). Total precision was also Mesoridazine 50 calculated from these data. Table 2 summarizes the data, which are presented in mAU/min.
2 9J052.4D_C Table 7 lists the concentrations of compounds that show a negative response to the 9 References Emit® II Plus Phencyclidine Assay at a 25 ng/mL cutoff level. Table 7 — Concentrations of Compounds Showing a Negative Response 1. Hawks RL, Chiang CN, eds. Urine Testing for Drugs of Abuse. Rockville, MD: National Institute on Drug Abuse (NIDA), NIDA research monograph 73. Department of Health and Human Compound Concentration (µg/mL) Services; 1986. Acetaminophen 1000 2. Aniline O, Pitts FN. Phencyclidine (PCP): A review and perspectives. CRC Crit Rev Toxicol. a-Acetyl-N,N-dinormethadol (dinor LAAM) 15 1982: 10:145–177. L-a-Acetylmethadol (LAAM) 25 3. Finkle HI. Phencyclidine identification by thin-layer chromatography. A rapid screening procedure for emergency toxicology. Am J Clin Pathol. 1978; 70:287–290. N-Acetylprocainamide (NAPA) 400 4. Froelich PE, Gross G. Separation and detection of phencyclidine in urine by gas Acetylsalicylic acid 1000 chromatography. J Chromatogr. 1977; 137:135–143. Amitriptyline 125 5. Gupta RC et al. Determination of phencyclidine in urine and illicit street drug samples. D-Amphetamine 1000 Clin Toxicol. 1975; 8:611–621. Benzoylecgonine 1000 6. Oellerich M. Enzyme immunoassays in clinical chemistry: Present status and trends. J Clin Chem Clin Biochem. 1980; 18:197–208. Buprenorphine 1000 Caffeine 1000 Chlorpromazine* Cimetidine 1000 Symbols Key Clomipramine 2.5 Clonidine 1000 Do not reuse
Codeine 500 Use By Cotinine 100 Batch Code Cyclobenzaprine 62 Desipramine 800 Catalogue Number Diphenhydramine 1000 Caution, consult accompanying documents Doxepin 250 2-Ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) 1000 Manufacturer Fluoxetine 1000 Authorized Representative in the European Community Glutethimide 500 Ibuprofen 1000 Contains sufficient for
Sensitivity The sensitivity level (minimum detection limit) of the Emit® II Plus Phencyclidine Assay is 1.4 ng/mL. This level represents the lowest concentration of phencyclidine that can be distinguished from 0 ng/mL with a confidence level of 95%.
Made in USA for Beckman Coulter, Inc. 250 S. Kraemer Blvd. Brea, CA 92821
® Revised August 2010 Printed in USA 9J052.4D_C