The Creative Approach to Bioscience

Total Protein (Single Reagent)

REF: 310 001 (2 x 100ml) 200 test
REF: 310 002 (4 x 100ml) 400 test
REF: 310 003 (8 x 100ml) 800 test
REF: 310 004 (2 x 500ml) 1000 test
REF: 310 005 (2 x 250ml) 500 test
REF: ZL-310 001 200 test

Intended Use Deterioration

Spectrum Total protein reagent is intended for the in-vitro quantitative, Do not use The total protein regents if precipitate forms. Failure to
diagnostic determination of total protein in human serum or plasma recover control values within the assigned range may be an indication
on both automated and manual systems. of reagent deterioration.

Background Specimen Collection and Preservation

Plasma proteins are mainly synthesized in the liver and are involved Use serum or plasma ( EDTA or heparin ) for the test . Usually
in the maintenance of normal water distribution between tissues and plasma results are higher due to fibrinogen . The serum or plasma
blood, as well as acid-base balance. Due to some pathological should be separated from the cells within 4 hours .
conditions, both total protein level and the ratio of different fractions Stability : 1 day at 15 – 25 oC ; 4 weeks at 4 – 8 oC;
may change independently of one another. Hyperproteinemia may 1 year at -20 oC
be detected during dehydration associated with diarrhea or vomiting.
The total protein levels also increase in multiple myeloma. System Parameters
Hypoproteinemia may occur as a result of prolonged low protein diet Wavelength Hg 546 nm (530 – 570 nm)
and in some pathological conditions such as nephrotic syndrome, Optical path 1 cm
bleeding, sprue, and salt retention. Assay type End-point
Direction Increase
Method Sample : Reagent Ratio 1 : 50
e.g .: Reagent volume 1 ml
Colorimetric method (Biuret reagent). Sample volume 20 l
Temperature 15 – 25 oC
Assay Principle Zero adjustment Reagent blank
Incubation time 10 minutes at 15 – 25 oC
In alkaline medium the copper reacts with the peptide bonds of Reagent Blank Limits Low 0.00 AU
proteins to form the characteristic pink to purple biuret complex. High 0.3 AU
Sodium potassium tartarate prevents copper hydroxide precipitation, Sensitivity 1.0 g/dL
and potassium iodide prevents the autoreduction of copper. Linearity 12 g/dL

Protein + Cu2+ Alkaline pH Cu – protein complex Procedure

The color intensity is directly proportional to the protein concentration.It Blank Standard Sample
is determined by measuring the increase in the absorbance at 546nm.
Reagent (R) 1.0 ml 1.0 ml 1.0 ml
Reagents Standard ------ 20 l ------
Sample ------ ------ 20 l
Standard Total protein (ST)
6.0 g/dL Mix and Incubate for 10 minutes at room temp. Measure absorbance
of specimen (Aspecimen) and standard (Astandard) against reagent
Reagent (R) blank within 30 minutes.
Sodium hydroxide 750 mmol/L
Copper sulfate 12.0 mmol/L Calculation
Sodium potassium tartarate 40.9 mmol/L Aspecimen
Potassium iodide 19.8 mmol/L Serum protein conc. (g/dL) = x6
Astandard
(C)-Corrosive contains caustic materials.
R34 Causes burns. Note: For turbid highly icteric sera, prepare a serum blank by adding
S26-45 In case of contact with eyes, rinse immediately with plenty 20 l serum to 1 ml saline into a labeled test tube. Read absorbance
of water and seek medical advice in case of accident or if of serum blank at 540 nm vs water and subtract serum blank
you feel unwell, seek medical advice immediately. absorbance from test absorbance before calculating results.
For further information, refer to the Total Protein reagent material Interfering Substances
safety data sheet.
Hemolysis
Precautions and Warnings No interference up to hemoglobin level of 7.5 g/L.
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Icterus
immediately with plenty of soap and water. In case of severe injuries; No significant interference up to a bilirubin level of 30 mg/dL.
seek medical advice immediately.
Lipemia
Reagent Preparation, Storage and Stability No significant interference.

Spectrum Total protein reagents are supplied ready-to-use and stable Drugs
up to the expiry date labeled on the bottles. The reagents are stable Sera from patients receiving dextran may cause artificially high levels
at 15 – 25 oC. Only the standard needs to be kept refrigerated at due to turbidity during color development. This positive bias can be
(2 - 8oC).Once opened, the reagent is stable for 6 months and the minimized by centrifuging the reaction mixture before reading the
standard is stable for 3 months at the specified temperature if absorbance.
contamination is avoided.
 Waste Disposal
Performance Characteristics
This product is made to be used in professional laboratories. Please
Precision consult local regulations for a correct waste disposal.
Within run (Repeatability) S56: dispose of this material and its container at hazardous or
special waste collection point.
Level 1 Level 2 S57: use appropriate container to avoid environmental contamination.
n 20 20 S61: avoid release in environment. refer to special instructions/safety
data sheets.
Mean (g/dL) 5.2 7.23
References
SD 0.12 0.15
CV% 2.31 2.07 1. Cannon DC, Olitzky I, Inkpen JA :Proteins. In:Clinical chemistry,
principles and technics, 2 nd ed. RJ Henery, DC Cannon, JW
Run to run (Reproducibility) Winkelman, editors, Harper & Row, New York, pp 407 – 421,1974.
2. Gornall AG, Bardawill CJ, David MM: Determination of serum
protein by means of the biuret reagent. J Biol Chem 177:751,
Level 1 Level 2 1949 .
3. Kaplan A, Szalbo J :Clinical chemistry :Interpretation and
n 20 20 techniques, 2 nd ed. A Kaplan, J Szabo, editors, 1983, p 157.
Mean (g/dL) 5.7 7.32 4. Schultze HE, Heremans JF:Molecular biology of human
protein. Elsevier publishing company, Amsterdam, 1966.
SD 0.19 0.21 5. Tietz NW : Fundamentals of Clinical Chemistry: 2 nd ed. NW Tietz,
editor, 1994, pp692 .
CV% 3.33 2.87
Quality Control
ORDERING INFORMATION
Normal and abnormal commercial control serum of known concentrations
should be analyzed with each run. CATALOG NO. QUANTITY

Methods Comparison 310 001 2 x 100 ml

310 002 4 x 100 ml
A comparison between Spectrum Total protein reagent and a commercial 310 003 8 x 100 ml
reagent of the same methodology was performed on 200 human sera. 310 004 2 x 500 ml
A correlation of 0.978 was obtained. 310 005 2 x 250 ml
REF: ZL-310 001 200 test
Sensitivity
When run as recommended, the minimum detection limit of this
assay is 1.0 g/dL.

Linearity
The reaction is linear up to total protein concentration of 12 g/dL.
Specimens showing higher concentration should be diluted 1+1
using physiological saline and repeat the assay (result × 2).

Expected Values
Adults 6.6 – 8.7 g/dL

Children (> 1 year) 6.0 – 8.0 g/dL

(< 1 year) 4.8 – 7.6 g/dL

Newborns (< 4 weeks) 4.6 – 6.8 g/dL

Prematures 3.4 – 5.0 g/dL

Analytical Range
1.0 – 12 g/dL.

Spectrum Diagnostics does not interpret the results of a clinical

laboratory procedure;interpretation of the results is considered
the responsibility of qualified medical personnel. All indications
of clinical significance are supported by literature references.

Egyptian Co for Biotechnology - Spectrum Diagnostics (S.A.E)

Obour city industrial area. block 20008 piece 19 A. Cairo. Egypt.
Tel: +202 4489 2248 - Fax: +202 4489 2247
www.spectrum-diagnostics.com
E-mail:info@spectrum-diagnostics.com

MDSS GmbH
EC REP Schiffgraben 41
30175 Hannover, Germany IFUFCC38 Rev.(6), 6/6/2021