Uric Acid liquicolorplus R1-R2 procedure:

Pipette into cuvettes Reagent blank (Rb) Sample or [STD]

PAP-Method Sample / [STD] --- 20 μl
Enzymatic Colorimetric Test [BUF] 1000 μl 1000 μl
Package Sizes Mix, incubate for approx. 1 minute.
[REF] 10694 3 x 100 ml Complete Test Kit [ENZ] 250 μl 250 μl
[IVD] Mix, incubate for 5 minutes at 37°C. Read absorbance ΔA against Rb
within 30 minutes.
Method1,2
Determination of uric acid by reaction with uricase. The formed H2O2 Procedure with [WR]:
reacts under catalysis of peroxidase with N-ethyl-N-(2-hydroxy-3-
Pipette into cuvettes Reagent blank (Rb) Sample or [STD]
sulfopropyl)-3-methylaniline sodium salt (TOOS) and 4-aminophenazone
(PAP) to give a red-violet quinoneimine dye as indicator. Potential interfe- Sample / [STD] --- 20 μl
rence by ascorbic acid is avoided by the integration of ascorbate oxidase in [WR] 1000 μl 1000 μl
the reagent. Mix, incubate for 5 minutes at 37°C. Read absorbance ΔA against Rb
within 30 minutes.
Reaction Principle
uricase Calculation of the Uric Acid Concentration
Uric acid + O2 + 2 H2O ⎯⎯⎯⎯→ allantoin + CO2 + H2O2 1. Serum, Plasma
C = 8 x ΔA Sample / ΔA [STD] [mg/dl] or
peroxidase C = 476 x ΔA Sample / ΔA [STD] [μmol/l]
2 H2O2 + TOOS + PAP ⎯⎯⎯⎯→ Quinoneimine + HCl + 4 H2O
2. Urine
Contents C = 88 x ΔA Sample / ΔA [STD] [mg/dl] or
[BUF] 3 x 80 ml Buffer C = 5235 x ΔA Sample / ΔA [STD] [μmol/l]
Phosphate buffer (pH 7.5) 100 mmol/l
TOOS 1 mmol/l Performance Characteristics
Ascorbate oxidase ≥ 1 KU/l Linearity: up to 25 mg/dl or 1488 μmol/l
[ENZ] 1 x 60 ml Enzymes Dilute samples with a higher concentration 1+1 with physiological saline
Phosphate buffer (pH 7.5) 100 mmol/l (0.9%). Multiply the results by 2. When used on instruments, the linear
4-Aminophenazone 0.3 mmol/l range depends on the respective application (refer to application sheets).
Potassium hexacyanoferrate (II) ≥ 10 μmol/l Typical performance data can be found in the Verification Report,
Peroxidase ≥ 1 KU/l accessible via
Uricase ≥ 0.1 KU/l
www.human.de/data/gb/vr/su-uracp.pdf or
[STD] 1 x 3 ml Standard
www.human-de.com/data/gb/vr/su-uracp.pdf
Uric acid 8 mg/dl
or 476 μmol/l Reference Values3
Reagent Preparation Men: 3.4 - 7.0 mg/dl or 200 - 420 μmol/l
The reagents are ready for use and can directly be applied on automated Women: 2.4 - 5.7 mg/dl or 140 - 340 μmol/l
analyzers (R1-R2 procedure). Urine: 250 - 750 mg/24h or 1.5 - 4.5 mmol/24h
[WR] (working reagent) is prepared by mixing 4 parts [BUF] with 1 part
[ENZ], e.g. 80 ml [BUF] + 20 ml [ENZ].
Quality Control
All control sera with uric acid values determined by this method can be
Reagent Stability employed. We recommend to use our quality control sera HUMATROL
The individual reagents are stable, even after opening, up to the stated based on animal serum or our SERODOS based on human serum.
expiry date when stored at 2...8°C. Contamination of the reagents must be
strictly avoided. Automation
Proposals to apply the reagents on analyzers are available on request.
[WR] is stable for 3 weeks at 15...25°C and for 3 months at 2...8°C.
Each laboratory has to validate the application in its own responsibility.
All reagents have to be stored protected from light.
Notes
Specimen 1. With R 1 – R 2 procedure the test is not influenced by hemoglobin (up
Serum, heparinised or EDTA-plasma, urine. to 500 mg/dl), bilirubin (up to 12 mg/dl), triglycerides (up to 2500
Dilute urine 1+10 with dist. water. mg/dl) and ascorbic acid (up to 20 mg/dl); with [WR] the test is not
influenced by hemoglobin (up to 50 mg/dl), bilirubin (up to 20 mg/dl),
Uric acid remains stable in serum for 3 to 5 days at 2...8°C, or for 6 months triglycerides interfere and ascorbic acid (up to 20 mg/dl).
at -20°C.
2. The reagents contain sodium azide (0.095%) as preservative. Do not
In urine uric acid remains stable for approximately 3 days at room tem- swallow. Avoid contact with skin and mucous membranes!
perature, provided bacterial contamination is avoided.
Note: Lipemic specimens usually generate turbidity of the sample References
reagent mixture which leads to falsely elevated results. The URIC 1. Barham, D., Trinder P., Analyst 97, 142 (1972)
ACID liquicolorPLUS test avoids these falsely elevated results 2. Fossati, P. et al., Clin. Chem. 26/2, 227 (1980)
through its built-in Lipid-Clearing Factor (LCF). The LCF clears up
3. Thefeld, L. et al., Dtsch. Med. Wschr. 98, 380-384 (1973)
totally a turbidity caused by lipemic specimens.

Assay
Wavelength: 520 nm, 546 nm SU-URACP INF 1069401 GB 12-2012-09
Optical path: 1 cm
Temperature: 37°C
Measurement: Against reagent blank (Rb). Only one reagent blank per
series is required.

Pipetting Scheme
Please use only the standard recommended by HUMAN (enclosed in the
kit).

Human Gesellschaft für Biochemica und Diagnostica mbH

Max-Planck-Ring 21 · 65205 Wiesbaden · Germany
Telefon +49 6122-9988-0 · Telefax +49 6122-9988-100 · e-Mail human@human.de