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All components are liquid and ready to use. Reagent preparation is not necessary.

For the quantitative in vitro determination of unsaturated iron binding capacity (UIBC) in serum.

Determination of serum iron binding capacity takes place in the first line of most widely used diagnostic tests for the evaluation of iron metabolism and investigation of iron metabolism related disorders. Approximately 30% capacity of serum transferrin level is occupied with iron at physiological conditions in healthy subjects. Serum unsaturated iron binding capacity

(UIBC) level is usually increased in iron deficiency anemia and decreased in hemochromatosis, malignancies, rheumatic fever and collagen vascular disease.

Principle

In the measurement of new generation assay of UIBC, a known amount of stabilized ferric iron is added, in excess, to the serum at alkaline pH. This saturates the unoccupied iron binding sites on the transferrin. The amount of unbound ferric ion is measured and then subtracted from the total quantity.

The New Generation UIBC Reagents, R1 and R2, and calibrator are ready to use as supplied. No reagent preparation is required.

Reagent 1 (R1) ; Alkaline buffer containing ferric ion 1X50 ml

Reagent2 (R2) ; Chromogen and reductant 1X 6 ml

Standard (Std) ; Liquid solution 1X 5 ml

The reagent is stable until the expiration date shown on the label when stored at 2-8°C. Do not freeze the reagents. Protect reagents from light.

Automated Procedure for Abbott Architect c8000 Analyzers

Reagent 1 (R1) volume: 250 µL

Sample volume: 15 µL

Reagent 2 (R2) volume: 30 µL

Wavelength (s): 600 nm/(as secondary; 700 nm)

Reading points: 13/33 The first absorbance is taken before the mixing of R1 and R2 (as sample blank) and the last absorbance is taken when the reaction trace draws a plateau line (about 3– 4 min after the mixing).

Calibration type: Linear

The assay can be performed in a variety of automated chemistry analyzers. Details available on request. All performance data included here were obtained using a Abbott Architect c8000 analyzer.

The instrument automatically calculates the results.

Concentration	Mean, g/dL	SD	CV%
Low	237.7	4.05	1.70
Medium	351.7	3.30	0.93
High	478.8	4.04	0.84

Non-hemolyzed, non-lipemic and non-icteric serum is the recommended sample. Do not use EDTA-plasma. Specimens should be collected in the morning in order to avoid low results due to diurnal variation. Samples should be separated from the blood cells and analyzed promptly. However, the serum may be stored at 2-8°C, or at -20°C for up to one month. Serum can be stored at room temperature (22-28°C) for two weeks. Discard contaminated specimens.

General laboratory equipment.

The new generation UIBC Kit is for in-vitro diagnostic use. Normal precautions for handling laboratory reagents should be taken. Do not mix reagents of different lot numbers. Universal Precautions, as they apply to your facility, should be used for handling and disposal of materials during and after testing. All specimens and controls being tested should be considered potentially infectious.

Do not ingest reagent, do not pipette by mouth. Prevent contact with skin and eyes. Avoid contact with skin and eyes. If spilt, thoroughly wash affected areas with water. For further information consult the Relassay New Generation UIBC Reagent Material Safety Data Sheet.

An included aqueous (or serum based) UIBC calibrator, with an assigned value is used. It is not necessary to determine a standard curve with this procedure since the reaction is linear to 750 µg/dL. A calibrator and reagent blank should be determined with each set of unknowns assayed. Follow the instrument manufacturer’s guidelines for calibration performance and frequency, using quality control samples with each run to verify satisfactory calibration. (Results expressed in µg/dL may be converted to µmol/L by multiplying by 0.1791).

Reliability of test results should be monitored by including control sera, with known UIBC concentrations, in each assay run. These controls should be carried through the process and treated in the same manner as the patient’s serum samples. The recovery of control values within the established acceptable range should be the criteria used in the evaluation of assay performance.

The New Generation UIBC method demonstrated linearity between 25- 750 µg/dL UIBC. When values exceed the upper limit samples should be diluted 1 + 1 with NaCl solution (9 g/L) and the result multiplied by 2.

The low limit of detection is 25 µg/dL. The limit of detection may be a function of the analyzer. Users should establish their own low detection limits.

A reference range of 70 - 380 µg/dL (12.5 - 68 µmol/L) for males and 70 - 390 µg/dL (12.5 - 70 µmol/L) for females was calculated from the serum iron and TIBC ranges reported in literature. These ranges should serve only as guidelines. It is recommended that each laboratory establish its own range of expected values, since differences exist between instruments, laboratories, and local populations.

Please refer to local legal requirements.

1.Tietz NW (ed). Textbook of Clinical Chemistry, 3rd ed. Philadelphia,

PA: WB Saunders; 1701-1703; 1999.

2.NCCLS. Determination of Serum Iron and Total Iron Binding Capacity;

Proposed Standard, NCCLS Document H17-P. Wayne, PA: NCCLS,

Vol. 10, No. 4; 1990.

3.Sidloi M, Koch P, Tonks L, Tonks DB. A fully automated method for determining total serum iron and unsaturated iron-binding capacity using a Technicon AA-II AutoAnalyser system. Clin Biochem. 1982 Apr;15(2):101-5.

4.Worwood M. The laboratory assessment of iron status--an update. Clin Chim Acta. 1997 Mar 18;259(1-2):3-23.

5.T.P.E. Patent Number 175858

MEGA TIP, Industry and Trade Co. Gaziantep, Turkey.

Address: Bahcelievler Mah. K.Ismailpasa Sk. No:10 Sahinbey / Gaziantep / TURKIYE

www.relassay.com

relassay@relassay.com

Phone : +90 342 230 37 52

Fax : +90 342 220 49 87

References

Waste Management

Reference Range

Low Detection Limit

Linearity

Quality Control

Calibration

WARNING:

Safety Precautions and Warnings

Materials Required but not Provided

Specimen Collection and Storage

Precision Studies

Calculation of Results

Automated Procedure

Reagent Storage and Stability

Reagents

Reagent Preparation

Excess iron(III) + (apo)transferrin alkaline pH Remaining iron + holotransferrin

Iron + Chromogen Colored complex

Introduction

Intended Use

Specification

New Generation UIBC Assay Kit