Professional Documents
Culture Documents
ALKALINE PHOSPHATASE
This package insert contains information to run the Alkaline Phosphatase assay on the ARCHITECT c Systems™
and the AEROSET System.
NOTE: This package insert must be read carefully prior to product use. Package insert instructions must be followed
accordingly. Reliability of assay results cannot be guaranteed if there are any deviations from the instructions in
this package insert.
Customer Support
United States: 1-877-4ABBOTT
Canada: 1-800-387-8378 (English speaking customers)
1-800-465-2675 (French speaking customers)
International: Call your local Abbott representative
Reagent 2
1
NAME WARNINGS AND PRECAUTIONS
ALKALINE PHOSPHATASE Precautions for Users
1. For in vitro diagnostic use.
INTENDED USE
2. Do not use components beyond the expiration date.
The Alkaline Phosphatase assay is used for the quantitation of alkaline
phosphatase in human serum or plasma. 3. Do not mix materials from different kit lot numbers.
4. is classified per applicable European Community (EC) Directives
SUMMARY AND EXPLANATION OF TEST as: Irritant (Xi). The following are the appropriate Risk (R) and Safety
(S) phrases:
Human alkaline phosphatase (AlkP, EC.3.1.3.1) consists of a group of at
least five tissue-specific isoenzymes which catalyzes the hydrolysis of R36/38 Irritating to eyes and skin.
phosphate mono-esters at alkaline pH. A variety of disease processes R52/53 Harmful to aquatic organisms, may cause long-
can result in the release of increased quantities of alkaline phosphatase term adverse effects in the aquatic environment.
into the blood. S26 In case of contact with eyes, rinse immediately
with plenty of water and seek medical advice.
PRINCIPLES OF PROCEDURE S35 This material and its container must be disposed
Several substrates have been used to measure alkaline phosphatase of in a safe way.
activity such as glycerophosphate,1 phenyl phosphate,1 and S37 Wear suitable gloves.
p-nitrophenyl phosphate.2 Bowers and McComb3 improved the method S46 If swallowed, seek medical advice immediately
of Bessey et al. to include a kinetic measurement. Tietz et al.4 and show this container or label.
optimized this method to include a chelated metal-ion buffer of zinc, S61 Avoid release to the environment. Refer to special
magnesium, and HEDTA. This Alkaline Phosphatase procedure is a instructions/safety data sheet.
modification of this method. For product not classified as dangerous per European Directive
Alkaline phosphatase in the sample catalyzes the hydrolysis of colorless 1999/45/EC as amended, safety data sheet available for
p-nitrophenyl phosphate (p-NPP) to give p-nitrophenol and inorganic professional user on request.
phosphate. At the pH of the assay (alkaline), the p-nitrophenol is in the 5. CAUTION: This product requires the handling of human specimens.
yellow phenoxide form. The rate of absorbance increase at 404 nm is It is recommended that all human sourced materials be considered
directly proportional to the alkaline phosphatase activity in the sample. potentially infectious and handled in accordance with the OSHA
Optimized concentrations of zinc and magnesium ions are present to Standard on Bloodborne Pathogens.5 Biosafety Level 26 or other
activate the alkaline phosphatase in the sample. appropriate biosafety practices7,8 should be used for materials that
Methodology: Para-nitrophenyl Phosphate contain or are suspected of containing infectious agents.
2
PROCEDURE RESULTS
Materials Provided Refer to the instrument-specific operations manual for information on
results calculations.
7D55-20 or 7D55-30 Alkaline Phosphatase Reagent Kit
• ARCHITECT System Operations Manual—Appendix C
Materials Required but not Provided • AEROSET System Operations Manual—Appendix A
• Control Material Representative performance data are given in the EXPECTED VALUES
• Saline (0.85% to 0.90% NaCl) for specimens that require dilution and SPECIFIC PERFORMANCE CHARACTERISTICS sections of this
Assay Procedure package insert. Results obtained in individual laboratories may vary.
For a detailed description of how to run an assay, refer to Section 5 of LIMITATIONS OF THE PROCEDURE
the instrument-specific operations manual.
Refer to the SPECIMEN COLLECTION AND HANDLING and SPECIFIC
Specimen Dilution Procedures PERFORMANCE CHARACTERISTICS sections of this package insert.
The ARCHITECT c Systems and the AEROSET System have automatic
dilution features; refer to Section 2 of the instrument-specific operations EXPECTED VALUES
manual for additional information.
Reference Range
Serum and plasma: Specimens with alkaline phosphatase values
exceeding 2,200 U/L (4,555 U/L for Flex Rate Linearity) are flagged Serum/Plasma12
and may be diluted using the Automated Dilution Protocol or the Manual
Dilution Procedure. Range (U/L)
Male
Automated Dilution Protocol
1 to 12 years < 500
If using the Automated Dilution Protocol, the system performs a dilution
of the specimen and automatically corrects the enzyme activity value 12 to 15 years < 750
by multiplying the result by the appropriate dilution factor. To set up the > 20 years 40 to 150
automatic dilution feature, refer to Section 2 of the instrument-specific Female
operations manual for additional information. 1 to 12 years < 500
Manual Dilution Procedure > 15 years 40 to 150
Manual dilutions should be performed as follows: It is recommended that each laboratory determine its own reference
• Use saline (0.85% to 0.90% NaCl) to dilute the sample. range based upon its particular locale and population characteristics.
• The operator must enter the dilution factor in the patient or control
order screen. The system uses this dilution factor to automatically SPECIFIC PERFORMANCE CHARACTERISTICS
correct the enzyme activity value by multiplying the result by the
entered factor. Linearity
Alkaline Phosphatase is linear up to 2,200 U/L.
• If the operator does not enter the dilution factor, the result must
be multiplied by the appropriate dilution factor before reporting the Flex Rate Linearity is 4,555 U/L. To use Flex Rate Linearity, the operator
result. must edit the linear high value to 4,555 on the appropriate screen.
NOTE: If a diluted sample result is flagged indicating it is less than the • ARCHITECT c Systems—Configure assay parameters screen,
linear low limit, do not report the result. Rerun using an appropriate Results view
dilution. • AEROSET System—Assay Configuration screen, Outline page
For detailed information on ordering dilutions, refer to Section 5 of the Linearity was verified using Clinical and Laboratory Standards Institute
instrument-specific operations manual. (CLSI) protocol NCCLS EP6-P.13
Limit of Detection (LOD)
CALIBRATION The LOD for Alkaline Phosphatase is 5.0 U/L. The LOD is the
Calibration is stable for approximately 10 days (240 hours) and is mean concentration of an analyte-free sample + 2 SD, where
required with each change in reagent lot number. Verify calibration SD = the pooled, within-run standard deviation of the analyte-free
with at least two levels of controls according to the established quality sample. A study performed on an ARCHITECT c System and an
control requirements for your laboratory. If control results fall outside AEROSET System produced an LOD for Alkaline Phosphatase
acceptable range, recalibration may be necessary. of 1.9 U/L.
A calibration factor (2150) must be entered.
Limit of Quantitation (LOQ)
• ARCHITECT c Systems—Configure assay parameters window,
Calibration view The LOQ for Alkaline Phosphatase is 5.0 U/L. The LOQ is the analyte
concentration at which the CV = 20%.
• AEROSET System—Assay Configuration screen, Calibration page
For a detailed description of how to calibrate an assay, refer to Interfering Substances14
Section 6 of the instrument-specific operations manual. Interference studies were conducted on the AEROSET System using
CLSI protocol NCCLS EP7-P.15 Interference effects were assessed by
QUALITY CONTROL Dose Response and Paired Difference methods, at the medical decision
level of the analyte.
The following is the recommendation of Abbott Laboratories for quality
control. As appropriate, refer to your laboratory standard operating Interfering Interferent Concentration N Target Observed
procedure(s) and/or quality assurance plan for additional quality control Substance (U/L) (% of Target)
requirements and potential corrective actions.
30 mg/dL (513 μmol/L) 4 155.6 102.4
• Two levels of controls (normal and abnormal) are to be run every Bilirubin
24 hours. 60 mg/dL (1,026 μmol/L) 4 155.6 102.7
• If more frequent control monitoring is required, follow the established 750 mg/dL (7.5 g/L) 4 150.8 95.3
quality control procedures for your laboratory. Hemoglobin
1,000 mg/dL (10.0 g/L) 4 150.8 94.3
• If quality control results do not meet the acceptance criteria 750 mg/dL (7.5 g/L) 4 144.3 100.4
defined by your laboratory, patient values may be suspect. Follow Intralipid
the established quality control procedures for your laboratory. 1,000 mg/dL (10.0 g/L) 4 144.3 100.2
Recalibration may be necessary. Bilirubin solutions at the above concentrations were prepared by addition
• Review quality control results and acceptance criteria following a of a bilirubin stock to human serum pools. Hemoglobin solutions at
change of reagent lot. the above concentrations were prepared by addition of hemolysate to
human serum pools. Intralipid solutions at the above concentrations were
prepared by addition of Intralipid to human serum pools.
3
SPECIFIC PERFORMANCE CHARACTERISTICS BIBLIOGRAPHY
(Continued) 1. King EJ, Armstrong AR. A convenient method for determining serum
and bile phosphatase activity. Can Med Assoc J 1934;31:376–81.
Precision
2. Bessey OA, Lowry OH, Brock MJ. A method for the rapid
The imprecision of the Alkaline Phosphatase assay is ≤ 6.2% Total CV.
determination of alkaline phosphatase with five cubic millimeters of
Representative data from studies using CLSI protocol NCCLS EP5-A16
serum. J Biol Chem 1946;164:321–9.
are summarized below.
3. Bowers GN, McComb RB. A continuous spectrophotometric method
Control Level 1 Level 2 for measuring the activity of serum alkaline phosphatase. Clin Chem
1966;12(2):70–89.
N 80 80 4. Tietz NW, Burtis CA, Duncan P, et al. A reference method for
Mean (U/L) 84.4 437.8 measurement of alkaline phosphatase activity in human serum.
Clin Chem 1983;29(5):751–6.
SD 3.89 3.17
Within Run 5. US Department of Labor, Occupational Safety and Health
%CV 4.6 0.7 Administration. 29 CFR Part 1910.1030. Occupational Exposure to
Bloodborne Pathogens.
SD 2.86 5.53
Between Run 6. US Department of Health and Human Services. Biosafety in
%CV 3.4 1.3 Microbiological and Biomedical Laboratories. HHS Publication
SD 0.00 7.29 (CDC), 4th ed. Washington, DC: US Government Printing Office,
Between Day May 1999.
%CV 0.0 1.7 7. World Health Organization. Laboratory Biosafety Manual. Geneva:
SD 4.83 9.68 World Health Organization; 2004.
Total 8. Sewell DL, Bove KE, Callihan DR, et al. Protection of Laboratory
%CV 5.7 2.2 Workers from Occupationally Acquired Infections; Approved
Guideline—Third Edition (M29-A3). Wayne, PA: Clinical and
Method Comparison Laboratory Standards Institute, 2005.
Correlation studies were performed using CLSI protocol NCCLS 9. Tietz NW, Rinker AD, Shaw LM. IFCC methods for the measurement
EP9-A.17 of catalytic concentration of enzymes, part 5: IFCC method for
Serum results from the Alkaline Phosphatase assay on the AEROSET alkaline phosphatase. J Clin Chem Clin Biochem 1983;21(11):
System were compared with those from a commercially available 731–48.
p-nitrophenyl phosphate hydrolysis methodology. 10. Guder WG, da Fonseca-Wollheim F, Heil W, et al. The Quality of
Serum results from the Alkaline Phosphatase assay on an ARCHITECT Diagnostic Samples. Darmstadt, Germany: GIT Verlag; 2001:14–5.
c System were compared with those from the Alkaline Phosphatase 11. US Pharmacopeial Convention, Inc. General notices. In: US
assay on the AEROSET System. Pharmacopeia National Formulary, 1995 ed. (USP 23/NF18).
AEROSET ARCHITECT vs. Rockville, MD: The US Pharmacopeial Convention, Inc; 1994:11.
vs. Comparative AEROSET 12. Burtis CA, Ashwood ER, editors. Tietz Textbook of Clinical
Method Chemistry, 2nd ed. Philadelphia, PA: WB Saunders; 1994:836.
13. Passey RB, Bee DE, Caffo A, et al. Evaluation of the Linearity
N 120 109 of Quantitative Analytical Methods; Proposed Guideline (EP6-P).
Y - Intercept -0.124 1.209 Villanova, PA: The National Committee for Clinical Laboratory
Standards, 1986.
Correlation Coefficient 0.999 1.000
14. Young DS. Effects of Drugs on Clinical Laboratory Tests, 4th ed.
Slope 1.059 0.981 Washington, DC: AACC Press; 1995:3-26-3-35.
Range (U/L)* 29.6 to 957.1 31.6 to 4,527.1 15. Powers DM, Boyd JC, Glick MR, et al. Interference Testing in
Clinical Chemistry; Proposed Guideline (EP7-P). Villanova, PA:
*AEROSET Range
The National Committee for Clinical Laboratory Standards, 1986.
16. Kennedy JW, Carey RN, Coolen RB, et al. Evaluation of Precision
Performance of Clinical Chemistry Devices; Approved Guideline
(EP5-A). Wayne, PA: The National Committee for Clinical
Laboratory Standards, 1999.
17. Kennedy JW, Carey NR, Coolen RB, et al. Method Comparison and
Bias Estimation Using Patient Samples; Approved Guideline (EP9-A).
Wayne, PA: The National Committee for Clinical Laboratory
Standards, 1995.
TRADEMARKS
AEROSET and ARCHITECT are registered trademarks of Abbott
Laboratories.
c System is a trademark of Abbott Laboratories.
All other trademarks, brands, product names, and trade names are the
property of their respective companies.
4
ARCHITECT c SYSTEMS ASSAY PARAMETERS
Rate linearity %: 10
Replicates: 3 [Range 1 – 3]
† The linear low value (Low-Linearity) is LOQ rounded up to the number of decimal places defined in the decimal places parameter field.
5
AEROSET SYSTEM ASSAY PARAMETERS
Refer to Assay Configuration in Section 2 of the AEROSET System Operations Manual for information regarding assay parameters.
* User defined or instrument defined.
** The linear low value (L-Linear Range) is LOQ rounded up to the number of decimal places defined in the decimal places parameter field.
‡‡ Rgt Name listed is for 7D55-20. For 7D55-30, change Reagent 1 name to ALKP051; Reagent 2 name remains ALKP052
6
7
8