Total Iron Ferrozine - Spectrum Diagnostics

1 Total iron - FerrozineREF: 272 001 (100 test)Reagent 1 2 x 45 mlReagent 1B 2 x 50 mlReagent 2 1 x 11 mlIntended UseSpectrum Diagnostics iron is intended for the in-vitro quantitative,diagnostic determination of Total iron in human majority of iron in the body (~3 g) is found in the haemoglobinof the red blood cells or their precursors in the bone marrow. Plasmacontains very small fraction of iron (~ mg). iron is transportedfrom one organ to another as a complex formed of ferric ions anda protein called apotransferrin, this iron -protein complex is calledtransferrin. The major iron -storage compound in the body is ferritin;it occurs in almost all body cells but particulary in iron is measured by the quantity of iron bound to transferrin,while TIBC is a direct measurement to transferrin. Elevated serumiron levels have been found in cases of hemochromatosis, hepatitis,hepatic necrosis and hemolytic anemia.

2 Decreased levels have beenassociated with iron defeciency anemia, chronic blood loss, chronicdisorders and insufficient dietary iron . The TIBC varies in disordersof iron metabolism so, TIBC is elevated in iron deffeciency measurements of both serum iron and TIBC is fundamental inevaluation and differential diagnosis of various types of anemia, liverdisease and chronic / Ferrozine PrincipleIronFerric ions are released from transferrin by guanidine hydrochlorideand reduced to ferrous state by hydroxylamine. Ferrous ions reactwith Ferrozine forming a coloured complex. To prevent copperinterference, cupric ions are bound to (III) Guanidine-HCL Apotransferrin +Fe(III)Fe(III) Hydroxylamine Fe(II)Fe(II) + Ferrozine Colored complexThe color intensity is directly proportional to the iron concentrationand is determined by monitoring the increase in absorbance at iron (ST) 200 mg/dL mmol/LReagent 1 (buffer pH )

3 Acetate buffer mol / LGuanidine hydrochloride mol / LHydroxylamine hydrochloride mol / LThiourea 100 mmol/LReagent 1B (buffer pH )Acetate buffer mol / LGuanidine hydrochloride mol / LHydroxylamine hydrochloride mol / LThiourea 100 mmol/LReagent 2: Ferrozine 60 mmol/LReagent PreparationPrepare the working solution by adding 5 ml of chromogen (R2) toone bottle of buffer (R1). Or prepare the working solution accordingto the number of tests required by mixing 9 volumes of R1 and1 volume of R2, 900 ml R1+100 and WarningsDo not ingest or inhalate.

4 In case of contact with eyes or skin; rinseimmediately with plenty of soap and water. In case of severe injuries;seek medical advice Storage and StabilityAll reagents are stable until expiration date stated on label whenstored refrigerated at 2 - 8 solution is stable for 6 months at 2 8 Collection and PreservationThe recommended specimen is serum or heparinized plasma. Plasmaspecimens collected with EDTA, oxalate, or citrate as anticoagulantsare unsatisfactory since they bind iron , preventing its reaction withthe chromogen. Morning specimen is preferrable to avoid low resultdue to diurinal variation. The biological half life of iron in blood isfew : 7 days at 15 25 oC ; 3 weeks at 2 8 oC; 1 year at -20 oCProcedure A-( iron ) Reagent Sample Blank Standard Blank SampleWorking ml ml ------ 200ml ------ ------ ------Standard ------ 200ml ------ ------Sample ------ ------ 200ml 200mlReagent 1B ------ ------ ml ------Mix, and incubate for 5 to 10 minutes at 20 25 oC.

5 Read theabsorbance of the standard and sample against reagent blank, andthe absorbance of sample blank against distilled water within 30minutes at 546 conc. (mg/dL) = x 200 Quality ControlNormal & abnormal commercial control serum of known concentrationsshould be analyzed with each - Asample blankAstandardIVDLOTREFoCoCEC REPSYMBOLS IN PRODUCT LABELLINGA uthorised RepresentativeFor in-vitro diagnostic useBatch Code/Lot numberCatalogue NumberConsult instructions for useTemperature LimitationUse by/Expiration DateCAUTION. Consult instructionsfor useManufactured byThe Creative Approach to BiosciencePerformance CharacteristicsPrecisionWithin run (Repeatiblity)Run to run (Reproducibility)Methods ComparisonA comparison between Spectrum Diagnostics iron reagent and acommercial reagent of the same methodology was performed on 20human sera.

6 A correlation of was run as recommended, the sensitivity of this assay is 5 mg/dLfor serum reaction is linear up to iron concentration of 500 mg/dL. Specimensshowing higher concentration should be diluted 1+1 using physiologicalsaline and repeat the assay (result 2).Interfering SubstancesSerum, plasmaHaemolysisNo interference up to haemoglobin level of 5 g/L ( mmol/L) indetermining serum iron and up to 1 g/L .IcterusNo significant interference up to a bilirubin level of 30 specimens are not recommended since they may causenegative bias. Lipemic specimens can be diluted before assay andthe dilution factor should be considered during , EDTA, and oxalate should be albumin levels more than 7 g/dL .Expected valuesIronNeonates : 36 184 mg/dL ( 6. 4 - 33 mmol/L)< 7 months: 37 145 mg/dL ( - 33 mmol/L)Adults Women: 37 145 mg/dL ( - 26 mmol/L)Men : 59 158 mg/dL ( - 28.)

7 Mmol/L)Analytical Range5 500 mg/dl ( mmol/ L).Waste DisposalThis product is made to be used in professional consult local regulations for a correct waste :dispose of this material and its container at hazardous or special waste collection :use appropriate container to avoid environmental :avoid release in environment. refer to special instructions/safetydata JD. Haemoglobin, porphyrin, and iron metabolism. In:KaplanLA, Pesce AJ, ed. Clinical Chemistry, theory, analysis, and correlation. ST. Louis:Mosby Company:1984 VF,Klee GG. Biochemical aspects of hematology. In : Tietz NW, ed. Fundamentals of clinical chemistry. 3rd ed. Philadelphia: WB saunders:1987 LL. Ferrozine -a new spectrophotometric reagent for Chem. 1970;42 MA, Gschwind H, Schl pfer P. Neue serumeisenbestimmungauf dem GSA II. Lab ;4 HL, Johnson DJ, Haut MJ. Simultaneous spectrophotometry of Fe2+ and Cu2+ in serum denatured with guanidine hydrochloride.

8 Clin ;23 Diagnostics does not interpret the results of a clinicallaboratory procedure; interpretation of the results is consideredthe responsibility of qualified medical personnel. All indicationsof clinical significance are supported by literature (mg/dL)SDCV% Total IronLevel (mg/dL)SDCV% Total iron ORDERING INFORMATIONCATALOG NO. QUANTITY 272 001 100 TestIFUFCC90 Rev.(1), 11/11/2013 MDSS GmbHSchiffgraben 4130175 Hannover, GermanyEC REPE gyptian Company for Biotechnology ( )Obour city industrial area. block 20008 piece 19 A. Cairo. : +202 4665 1848 - Fax: +202 4665