ALP STABILITY AND PREPARATION OF REAGENTS

R1a. Buffer
Alkaline Phosphatase Contents ready for use. Stable up to the expiry date
MANUAL when stored at +2 to +8C.
RX MONZA
R1b. Substrate
INTENDED USE Reconstitute one vial of Substrate R1b with the 10 ml
For the quantitative in vitro determination of Alkaline of Buffer R1a.
Phosphatase (ALP) in serum and plasma. This product is
suitable for manual use and on the RX monza analyser. Stable for 30 days at +2 to +8C or 3 days at +15 to
+25C.
Cat. No.
AP 307 R1a. Buffer 1 x 105 ml MATERIALS PROVIDED
10 x 10 ml R1b. Substrate 10 x 10 ml Buffer
Substrate
GTIN: 05055273200317
MATERIALS REQUIRED BUT NOT PROVIDED
COLORIMETRIC METHOD (1) Assayed Multi-sera Level 2 (Cat. No. HN 1530) and Level 3
This is an optimized standard method according to the (Cat. No. HE 1532)
recommendations of the Deutsche Gesellschaft für Klinische Randox Calibration Serum Level 3 (Cat. No. CAL 2351)
Chemie. RX series Saline (Cat. No. SA 3854)

PRINCIPLE PROCEDURE
ALP Aspirate fresh ddH2O and perform a new Gain Calibration in
p-nitrophenylphosphate + H2O phosphate + flow cell mode. Select ALP in the Run Test screen and carry
p-nitrophenol
out a water blank as instructed.
SAMPLE(2)
Serum or heparinized plasma. Pipette into a test tube:
Samples are stable for 5 days when stored at +2 to +8C.
Sample 0.01 ml
REAGENT COMPOSITION Reagent 0.5 ml

Contents Concentrations in the Test Mix and aspirate into the RX monza.
R1a. Buffer CALIBRATION FOR RX MONZA
Diethanolamine buffer 1 mol/l, pH 9.8 The use of Saline and Randox Calibration Serum Level 3 is
MgCl2 0.5 mmol/l recommended for calibration. Calibration is recommended
R1b. Substrate with change in reagent lot or as indicated by quality control
p-nitrophenylphosphate 10 mmol/l procedures.

FOR MANUAL USE

SAFETY PRECAUTIONS AND WARNINGS
For in vitro diagnostic use only. Do not pipette by mouth.
Wavelength: Hg 405 nm
Exercise the normal precautions required for handling Cuvette: 1 cm light path
laboratory reagents.
Temperature: 25C, 30C, 37C
Solution R1a contains Sodium Azide. Avoid ingestion or Measurement: against air
contact with skin or mucous membranes. In case of skin
contact, flush affected area with copious amounts of water. In Pipette into cuvette: Macro Semi- Micro
case of contact with eyes or if ingested, seek immediate Micro
medical attention.
Sample 0.05 ml 0.02 ml 0.01 ml
Sodium Azide reacts with lead and copper plumbing, to form Reagent (25C, 30C, 37C) 3.00 ml 1.00 ml 0.50 ml
potentially explosive azides. When disposing of such reagents,
flush with large volumes of water to prevent azide build up. Mix, read initial absorbance and start timer simultaneously.
Exposed metal surfaces should be cleaned with 10% sodium Read again after 1, 2 and 3 min.
hydroxide.
MANUAL CALCULATION
Solution R1a contains diethanolamine which may cause serious To calculate the ALP activity, use the following formulae:
damage to eyes and which is harmful if swallowed. Avoid
ingestion and wear suitable eye protection. U/l = 3300 x A 405 nm/min MACRO
U/l = 2760 x A 405 nm/min SEMI-MICRO
Health and Safety data sheets are available on request. U/l = 2760 x A 405 nm/min MICRO

The reagents must be used only for the purpose

intended by suitably qualified laboratory personnel,
under appropriate laboratory conditions.

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 MANUAL/ RX MONZA AP 307

QUALITY CONTROL PRECISION

Randox Assayed Multi-sera, Level 2 and Level 3 are recommended
for daily quality control. Two levels of controls should be assayed Intra Assay
at least once a day. Values obtained should fall within a specified Level 2 Level 3
range. If these values fall outside the range and repetition excludes Mean (U/l) 262 486
error, the following steps should be taken: SD 8.11 9.49
1. Check instrument settings and light source. CV (%) 3.10 1.95
2. Check cleanliness of all equipment in use. n 20 20
3. Check water. Contaminants i.e. bacterial growth may
contribute to inaccurate results. Inter Assay
4. Check reaction temperature. Level 2 Level 3
5. Check expiry date of kit and contents. Mean (U/l) 262 486
6. Contact Randox Laboratories Technical Services, SD 11.59 17.01
Northern Ireland +44 (0) 28 94451070. CV (%) 4.43 3.50
n 20 20
INTERFERENCE
Avoid haemolysis as it interferes with the assay. CORRELATION
The following analytes were tested up to the following levels This method (Y) was compared with another commercially
and were found not to interfere: available method (X) and the following linear regression
equation obtained:
Bilirubin 300 µmol/l (17 mg/dl)
Intralipid® 2% Y = 1.076X - 14.5
Triglycerides 22.75 mmol/l (2010 mg/dl) and a correlation coefficient of r = 0.9975
Haemoglobin 1 g/l (100 mg/dl)
43 patient samples were analyzed spanning the range 52 to
Physiological changes in serum or plasma analyte 965 U/l.
concentrations can be caused by a number of substances.
Comprehensive discussion of possible interfering substances, REFERENCES
their serum or plasma concentrations, and their possible 1. Rec. GSCC (DGKC); J. Clin. Chem. Clin. Biochem. 1972;
physiological involvements is beyond the scope of this 10: 182.
document. The listed reference contains specific details on 2. Englehardt A., et al, Aerztl Labor 1970 16 42.
known potential interfering substances (3). The user must 3. Young DS. Effects of Drugs on Clinical Laboratory Tests.
remain vigilant to the possible effect on results of unknown 5th ed. Washington, DC: AACC Press; 2000.
interferences from medications or endogenous substances. All
patient results must be evaluated in light of the total clinical The presence of a vertical bar in the margin indicates a technical
status of the patient. update from the previous revision.

NORMAL VALUES IN SERUM

25C 30C 37C

Men/women 60-170 U/l 73-207 U/l 98-279 U/l

It is recommended that each laboratory establish its own

reference range to reflect the age, sex, diet and geographical
location of the population.

SPECIFIC PERFORMANCE CHARACTERISTICS

The following performance data were obtained using a RX
monza analyser running at a temperature of 37C.

SENSITIVITY
The minimum detectable concentration of ALP with an
acceptable level of precision was determined as 49.9 U/l.

LINEARITY
This method is linear up to 1609 U/l. If the sample
concentration exceeds this value, dilute the sample 1+9 with Randox Teoranta, Meenmore,
0.9% NaCl solution and reassay. Multiply the result by 10. Dungloe, Donegal,
F94 TV06, Ireland

Revised 15 Dec 20 bm
Rev. 004

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