DIALAB Produktion und Vertrieb von chemisch – technischen Produkten und Laborinstrumenten Gesellschaft m.b.H.

A – 2351 Wiener Neudorf, Austria, IZ-NÖ Süd, Hondastrasse, Objekt M55

Phone: ++43 (0) 2236 660910-0, Fax: ++43 (0) 2236 660910-30, e-mail: office@dialab.at

Liquid Reagents – ready to use

REAGENT STABILITY AND STORAGE
Conditions: Protect from light.
PROTEIN TOTAL Close immediately after use.
Biuret Do not freeze the reagents.
2 Reagents Avoid contamination.
Substrate Start:
Diagnostic reagent for quantitative in vitro determination of Storage: at 2 – 25 °C
total protein in human serum or plasma on photometric Stability: up to the expiration date
systems Sample Start (Working Reagent):
Stability: at 2 – 25 °C 1 year
REF Kit Size Content SAMPLE STABILITY AND STORAGE
D95682B 1 x 12.5 L 1 x 10 L R1+ 2.5 L R2 Stability [3]: at 20 – 25 °C 6 days
D03120B 1 x 1.25 L 1 x 1 L R1 + 1 x 250 mL R2 at 4 - 8 °C 4 weeks
D95680 5 x 100 mL 4 x 100 mL R1 + 1 x 100 mL R2 at -20 °C at least 1 year
D00685 5 x 50 mL 4 x 50 mL R1 + 1 x 50 mL R2 Discard contaminated specimens.
D00686 5 x 25 mL 4 x 25 mL R1 + 1 x 25 mL R2 Freeze only once!
D00687 5 x 10 mL 4 x 10 mL R1 + 1 x 10 mL R2
D80911 10 x 50 mL 10 x 40 mL R1 + 4 x 25 mL R2 MATERIALS REQUIRED BUT NOT PROVIDED
D0437917 5 x 62.5 mL 4 x 62.5 mL R1 + 1 x 62.5 mL R2 NaCl solution (9 g/L)
DA0841 5 x 50 mL 5 x 40 mL R1 + 5 x 10 mL R2 General laboratory equipment
DT1041 4 x 62.5 mL 4 x 50 mL R1 + 4 x 12.5 mL R2
DK0738 5 x 50 mL 4 x 50 mL R1 + 1 x 50 mL R2 STANDARD
DB0941 2 x 150 mL 2 x 120 mL R1 + 2 x 30 mL R2 (not included in the kit – has to be ordered separately)
Concentration 5 g/dL (50 g/L)
Additionally available: Storage: 2 – 8 °C
D94683 1 x 3 mL Protein Total Standard Stability: up to the expiration date
D98485 5 x 3 mL Calibrator Diacal Auto Close immediately after use! Avoid contamination!
D98485SV 1 x 3 mL Calibrator Diacal Auto
D98481 12 x 5 mL Control normal Diacon N MANUAL TEST PROCEDURE
D14481 5 x 5 mL Control normal Diacon N Bring reagents and samples to room temperature.
D98481SV 1 x 5 mL Control normal Diacon N Substrate start
D98482 12 x 5 mL Control abnormal Diacon P Pipette into test tubes Blank Std./Cal. Sample
D14482 5 x 5 mL Control abnormal Diacon P
Sample - - 20 µL
D98482SV 1 x 5 mL Control abnormal Diacon P
Standard/Calibrator - 20 µL -
TEST PARAMETERS
Distilled water 20 µL - -
Method: Colorimetric, Endpoint, Increasing Reaction,
Biuret Reagent 1 1000 µL 1000 µL 1000 µL
Wavelength: 540 nm, Hg 546 nm Mix, read absorbance A1 against the reagent blank after 1 – 5
min. at 20-25 °C / 37 °C, than add:
Temperature: 20 – 25 °C, 37 °C
Sample: Serum or plasma Reagent 2 250 µL 250 µL 250 µL
Linearity: up to 15 g/dL (150 g/L) Mix, incubate for 5 min. at 20 – 25 °C / 37 °C and read
absorbance A2 against the reagent blank within 60 min.
Sensitivity: Lower limit of detection: 0.05 g/dL (0.5 g/L)
A = (A2 – A1) sample or std/cal.
SUMMARY [1,2] Sample start
Measurement of total protein is a useful test in a variety of Pipette into test tubes Blank Std./Cal. Sample
disorders. Decreased total protein concentrations can be
Sample - - 20 µl
detected in defective protein synthesis in the liver, protein loss
due to impaired kidney function, intestinal malabsorption or Standard/Calibrator - 20 µl -
nutritional deficiency. Elevated protein levels occur in chronic Dist. water 20 µl - -
inflammatory disorders, liver cirrhosis and dehydration. Reagent 1000 µl 1000 µl 1000 µl
TEST PRINCIPLE Mix, incubate for 5 min. at 20-25°C/37°C and read
Photometric test according to the Biuret method. absorbance against the reagent blank within 60 min.
Proteins form a violet blue colour complex with copper ions in
alkaline solution. CALCULATION
The absorbance of this colored complex is directly proportional A Sample
Total protein [g/dL] = x Conc. Std/Cal [g/dL]
to the protein concentration in the sample. A Std/Cal
REAGENT COMPOSITION UNIT CONVERSION
COMPONENTS CONCENTRATION g/dL x 10 = g/L
Reagent 1
Sodium hydroxide 100 mmol/L REFERENCE RANGE [1] * [g/dL]
Potassium sodium tartrate 17 mmol/L Females Males
Reagent 2 Adults: 6.6 - 8.8 6.6 - 8.8
Sodium hydroxide 500 mmol/L Children: Females Males
Potassium sodium tartrate 80 mmol/L
1 - 30 day(s) 4.2 - 6.2 4.1 - 6.3
Potassium iodide 75 mmol/L
1 – 6 month(s) 4.4 - 6.6 4.7 - 6.7
Copper sulphate 30 mmol/L
6 months – 1 year 5.6 - 7.9 5.5 - 7.0
REAGENT PREPARATION 1 – 18 year(s) 5.7 - 8.0 5.7 - 8.0
Substrate Start: *Each laboratory should check if the reference ranges are transferable to
Reagents are ready to use. its own patient population and determine own reference ranges if
Sample Start: necessary.
Mix 4 parts of Reagent 1 with 1 part of Reagent 2.
(= Working Reagent).

S:\pm\allg\Inserts_PM\Inserts_word\clinical chemistry\inserts dialab\protein total_en_rev05.docx Page 1 of 2 DI M. Wagner Rev. 05, 01.09.2016

 DIALAB Produktion und Vertrieb von chemisch – technischen Produkten und Laborinstrumenten Gesellschaft m.b.H.
A – 2351 Wiener Neudorf, Austria, IZ-NÖ Süd, Hondastrasse, Objekt M55
Phone: ++43 (0) 2236 660910-0, Fax: ++43 (0) 2236 660910-30, e-mail: office@dialab.at

PERFORMANCE CHARACTERISTICS P305+P351+P338: In in eyes: Rinse cautiously with water

LINEARITY, MEASURING RANGE for several minutes. Remove contact lenses, if present and
The test has been developed to determine total protein easy to do. Contrinue rinsing.
concentrations within a measuring range from 0.05 – 15 g/dL P337+P313: if eye irritation persists: Get medical
(0.5 – 150 g/L). advice/attention.
If values exceed this range samples should be diluted 1 + 1 with 3. The reagents contain sodium hydroxide. Do not swallow! If
NaCl solution (9 g/L) and the result multiplied by 2. the reagents come in contact with skin or mucous
SENSITIVITY/LIMIT OF DETECTION membranes rinse immediately with water!
The lower limit of detection is 0.05 g/dL (0.5 g/L). 4. The Protein Total Standard contains biological material.
The standard should be handled as potentially infectious
PRECISION (at 37 °C) and with the same precautions used for patient specimens.
Intra-assay Mean SD CV 5. In serum or plasma from patients who have received large
n = 20 [g/dL] [g/dL] [%] intravenous amounts of polydextrans too high values can
Sample 1 5.27 0.05 0.91 be measured with the biuret method. In such cases an
Sample 2 7.05 0.07 1.01 alternative method (e.g. Kjeldahl) has to be used.
Sample 3 10.4 0.08 0.80 6. In very rare cases, samples of patients with gammopathy
Inter-assay Mean SD CV might give falsified results [5].
n = 20 [g/dL] [g/dL] [%] 7. Please refer to the safety data sheets and take the
Sample 1 5.24 0.06 1.06 necessary precautions for the use of laboratory reagents.
Sample 2 7.07 0.11 1.53 8. For diagnostic purposes, the results should always be
Sample 3 10.4 0.14 1.32 assessed with the patient’s medical history, clinical
examinations and other findings.
SPECIFICITY/INTERFERENCES 9. For professional use only!
no interference up to:
Ascorbic acid 30 mg/dL WASTE MANAGEMENT
Bilirubin 40 mg/dL Please refer to local legal requirements.
Hemoglobin 500 mg/dL REFERENCES
Triglycerides 1000 mg/dL st
1. Thomas L. Clinical Laboratory Diagnostics. 1 ed.
Dextran 2000 mg/dL
Frankfurt: TH-Books Verlagsgesellschaft; 1998. p. 644-7.
For further information on interfering substances refer to Young
2. Johnson Am, Rohlfs EM, Silverman LM. Proteins. In:
DS [4].
Burtis CA, Ashwood ER, editors. Tietz Textbook of Clinical
rd
METHOD COMPARISON Chemistry. 3 ed. Philadelphia: W.B Saunders Company;
A comparison between Dialab Total protein (y) with a 1999. p. 477-540.
commercially available test (x) using 68 samples gave following 3. Guder WG, Zawta B et al. The Quality of Diagnostic
st
results: y = 1.00 x – 0.07 g/dL; r= 0.997. Samples. 1 e. Darmstadt: GIT Verlag; 2001; p.42-3.
th
CALIBRATION 4. Young DS. Effects of Drugs on Clinical laboratory Tests. 5
ed. Volume 1 and 2. Washington, DC: The American
The assay requires the use of a protein total standard or Association for Clinical Chemistry Press 2000.
calibrator.
5. Bakker AJ, Mücke M. Gammopathy interference in clinical
We recommend the Dialab Protein Total Standard and the
chemistry assays: mechanisms, detection and prevention.
Dialab multi calibration serum Diacal Auto. ClinChemLabMed 2007; 45(9): 1240-1243.
The assigned values of the calibrator are traceable to the Biuret
method.
QUALITY CONTROL
All control sera with protein total values determined by this
method can be used.
We recommend the Dialab serum controls Diacon N (control
serum with values in the normal range) and Diacon P (control
serum with values in the abnormal range).
Each laboratory should establish corrective action in case of
deviations in control recovery.
AUTOMATION
Special applications for automated analysers can be made on
request.
WARNINGS AND PRECAUTIONS
1. Reagent 1: Warning.
H290: May be corrosive to metals.
P234: Keep only in original container.
P390: Absorb spillage to prevent material damage.
2. Reagent 2: Warning.
H290: May be corrosive to metals.
H315: Causes skin irritation.
H319: Causes serious eye irritation.
H412: Harmful to aquatic life with long lasting effects.
P234: Keep only in original container.
P264: Wash hands and face thoroughly after handling.
P273: Avoid release to the environment.
P280: Wear protective gloves/protective clothing/eye
protection/face protection.
P332+P313: If skin irritation occurs: Get medical
advice/attention.

S:\pm\allg\Inserts_PM\Inserts_word\clinical chemistry\inserts dialab\protein total_en_rev05.docx Page 2 of 2 DI M. Wagner Rev. 05, 01.09.2016