Lab Policies Alkaline Phosphatase C311 Lab 8803

Standard Operating Procedure
Subject Alkaline Phosphatase C311

Index Number Lab-8803
Section Laboratory
Subsection Regional/Affiliates
Category Departmental
Contact Dayton, Judy L
Last Revised 9/17/2019
References
Required document for Laboratory Accreditation by the College of American Pathologists (CAP), Centers
for Medicare and Medicaid Services (CMS), and/or COLA.
Applicable To
Employees of the Gundersen Boscobel Area Hospitals and Clinics laboratories and Palmer Lutheran
Hospital and Clinics Laboratories.
Detail
PRINCIPLE:
The COBAS C311 provides a colorimetric assay in accordance with a standardized method.
In the presence of magnesium and zinc ions, p-nitrophenyl phosphate is cleaved by phosphatases into
phosphate and p-nitrophenol.
p-nitrophenyl phosphate + H2O → phosphate + p-nitrophenol

The p-nitrophenol released is directly proportional to the catalytic ALP activity. It is determined by
measuring the increase in absorbance.
CLINICAL SIGNIFICANCE:
Alkaline phosphatase in serum consists of four structural genotypes: the liver-bone-kidney type, the
intestinal type, the placental type and the variant from the germ cells. It occurs in osteoblasts,
hepatocytes, leukocytes, the kidneys, spleen, placenta, prostate and the small intestine. The liver-bone-
kidney type is particularly important.
A rise in the alkaline phosphatase occurs with all forms of cholestasis, particularly with obstructive
jaundice. It is also elevated in diseases of the skeletal system, such as Paget’s disease,
hyperparathyroidism, rickets and osteomalacia, as well as with fractures and malignant tumors. A
considerable rise in the alkaline phosphatase activity is sometimes seen in children and juveniles. It is
caused by increased osteoblast activity following accelerated bone growth.
SPECIMEN:
Universal precautions apply.
Serum and/or Li-heparin plasma
Specimen Stability:
7 days at 15-25 °C
Lab-8803 |Alkaline Phosphatase C311 Page 1 of 4

7 days at 2-8 °C
2 months at (-15)-(-25) °C
REAGENTS/MATERIALS:
COBAS C311
R1 2-amino-2-methyl-1-propanol: 1.724 mol/L, pH 10.44 (30 °C); magnesium acetate: 3.83 mmol/L;
zinc sulfate: 0.766 mmol/L; N-(2-hydroxyethyl)-ethylenediamine triacetic acid: 3.83 mmol/L
R2 p-nitrophenyl phosphate: 132.8 mmol/L, pH 8.50 (25 °C); preservatives
R1 is in position B and R2 is in position C.
Reagent precautions:
R1 contains 2-Amino-2-methyl-1-propanol.
Irritant
Irritating to eyes and skin.
Avoid contact with skin and eyes.
Reagents Stability:
ALP2S, ALP2L
Shelf life at 2-8 °C: See expiration date on COBAS c pack label.
On-board in use and refrigerated on the analyzer: 8 weeks
Diluent NaCl 9 %
Shelf life at 2-8 °C: See expiration date on COBAS c pack label.
On-board in use and refrigerated on the analyzer: 12 weeks
EQUIPMENT/INSTRUMENTATION:
Calibration:
Calibrators S1: H2O
S2: C.f.a.s.
Calibration mode Linear
Calibration frequency 2-point calibration
1. At initial installation
2. after reagent lot change
3. If required after maintenance or repair
4. as required following quality control procedures
Calibration Preparation, storage and stability: Refer to the COBAS C311 User Manual located in the
laboratory.
QUALITY CONTROL:
Quality Control frequency
Two levels of Quality control should be performed at a minimum:
1. once every twenty-four (24) hours

2. if a new cassette of reagent is put in use

3. if a calibration is performed
Refer to Lab-4405 Quality Control Criteria for Chemistry for interpretation of QC.
Implementation
Refer to the C311 User’s Manual located in the laboratory.
PROCEDURE NOTES:
AMR (Analytic Measurement Range): 5-1200 U/L
Extended Range:
5-6000 U/L with automatic re-run (1:5 dilution)
Report values less than 5 as “<5 U/L”

Report values greater than 6000 as “>6000 U/L”
If the provider requires further dilution on a sample reporting >6000 U/L, use 0.9% NaCl as the diluent.
Diluted value must fall between 5-1200 U/L.
Determine samples having higher activities via the rerun function. Dilution of samples via the rerun
function is a 1:5 dilution. Results from samples diluted using the rerun function are automatically
multiplied by a factor of 5.
CALCULATIONS: N/A
INTERPRETATION:
Expected Values
Adults: Males 40-129 U/L

Females 35-104 U/L
MALES
1 - < 10 years 142-335 U/L (2.37-5.59 µkat/L)
10 - < 13 years 129-417 U/L (2.15-6.96 µkat/L)
13 - < 15 years 116-468 U/L (1.94-7.82 µkat/L)
15 - < 17 years 82-331 U/L (1.37-5.53 µkat/L)
17 - < 19 years 55- 149 U/L (0.92-2.49 µkat/L)
FEMALES
1 - < 10 years 142-335 U/L (2.37-5.59 µkat/L)
10 - < 13 years 129-417 U/L (2.15-6.96 µkat/L)
13 - < 15 years 57-254 U/L (0.95-4.24 µkat/L)
15 - < 17 years 50-117 U/L (0.84-1.95 µkat/L)
17 - < 19 years 45-87 U/L (0.75-1.45 µkat/L)

LIMITATIONS:
Criterion: Recovery within ± 10 % of initial value at an alkaline phosphatase activity of 100 U/L (1.67
µkat/L).
Icterus: No significant interference up to an I index of 60 for conjugated and unconjugated bilirubin
(approximate conjugated and unconjugated bilirubin concentration: 1026 µmol/L or 60 mg/dL).
Hemolysis: No significant interference up to an H index of 200 (approximate hemoglobin concentration:

124 µmol/L or 200 mg/dL).
Lipemia (Intralipid): No significant interference up to an L index of 2000. There is poor correlation

between the L index (corresponds to turbidity) and triglycerides concentration.
Drugs: No interference was found at therapeutic concentrations using common drug panels.
In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause
unreliable results.
For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical
history, clinical examination and other findings.
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations
are run together on Roche/Hitachi cobas c systems. The latest version of the carry-over evasion list can
be found with the NaOHD/SMS/Multiclean/SCCS or the NaOHD/SMS/SmpCln1+2/SCCS Method Sheets.
For further instructions refer to the operator's manual. Where required, special wash/carry-over evasion
programming must be implemented prior to reporting results with this test.
REVIEW AND CHANGES:

This document and all attached forms should be reviewed optimally on an annual basis, with two years
as the maximum review date. Review will be done by the Technical Leader, Supervisor, Manager,
Medical Director or other designated person. Changes require retyping the document, and review by
the Medical Director.
REFERENCES:
1. COBAS C311 Alkaline Phosphatase IFCCmethod Sheet, Roche Doagnostics, Indianapolis, IN
2. Moss, DW; Alkaline Phosphatase isoenzymes; Clin Chem 1982; 28: 2007-2016
3. Hausamen TU, Helger R, Rick W, et al. Optimal conditions for the determination of serum alkaline
phosphatase by a new kinetic method. Clin Chim Acta 1967;15:241-245.
4. Tietz NW, Rinker AD, Shaw LM. J Clin Chem Clin Biochem 1983;21:731-748.
5. COBAS C311 User’s Manual.
6. COBAS C311 Method Sheet.

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Standard Operating Procedure

Subject Alkaline Phosphatase C311

p-nitrophenyl phosphate + H2O → phosphate + p-nitrophenol

Serum and/or Li-heparin plasma

Lab-8803 |Alkaline Phosphatase C311 Page 1 of 4

R2 p-nitrophenyl phosphate: 132.8 mmol/L, pH 8.50 (25 °C); preservatives

R1 is in position B and R2 is in position C.

Lab-8803 |Alkaline Phosphatase C311 Page 2 of 4

2. if a new cassette of reagent is put in use

Report values less than 5 as “<5 U/L”

Adults: Males 40-129 U/L

Lab-8803 |Alkaline Phosphatase C311 Page 3 of 4

Hemolysis: No significant interference up to an H index of 200 (approximate hemoglobin concentration:

Lipemia (Intralipid): No significant interference up to an L index of 2000. There is poor correlation

REVIEW AND CHANGES:

Lab-8803 |Alkaline Phosphatase C311 Page 4 of 4

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