ACCUCARETM
HEMOGLOBIN
(Cyanomet hemoglobin)
ORDER INFORMATION
Catalog No: Product Description: Pack Size:
HM 1000 HEMOGLOBIN 1 X 1000 ML
HM 5000 HEMOGLOBIN 1 X 5000 ML

METHOD
Colorimetric endpoint test

PRINCIPLE
In the cyanomethemoglobin method, erythrocytes are lysed by a stromatolytic reagent in the presence of a surfactant and release their hemoglobin into solution. Hemoglobin is oxidized to methemoglobin by ferricyanide, and the methemoglobin is converted into the stable cyanomethemoglobin by addition of KCN. The absorbance of cyanomethemoglobin is measured at 540 nm and colour intensity is proportional to hemoglobin concentration.

SAMPLE:
Collect whole blood with EDTA using aseptic technique. Whole blood collected with EDTA is stable for one week
at 2 - 8°C.

REFERENCE VALUES
Adult Males 13.0 - 18.0 g/dl
Adult Females 11.0 - 16.0 g/dl
Children 10.0 - 14.0 g/dl
Newborns 14.0 - 23.0 g/dl

REAGENTS
Reagent I : Cyanomethemoglobin reagent
Methemoglobin Std : 60 mg/dl (15 g/dl) store at 2-8°C

Interfering substances
Falsely elevated values are found in Lipaemic samples, Abnormal plasma proteins etc. Numerous drugs exert an in vivo effect to decrease blood hemoglobin.

REFERENCES
Eilers R.J., Am. J. Clin Path., 47 : 212 (1967).
Teitz N.W.,Fundamentals of clinical chemistry.
2nded.W.B.Saunders Co., Philadelphia p 411 (1976)
REAGENT PREPARATION
The Reagents are ready to use.

STABILITY : The stability is Till Expiry when stored at R.T. (away from sunlight).
The standard is stable at 2 - 8°C.

AUTOMATED PARAMETERS
Wave length 540 nm
Cuvette 1 cm light path
Reaction Temperature R.T.
Measurement Against reagent blank
Reaction Type End Point
Sample/Reagent Ratio 1 : 250
Incubation 5 minutes
Low Normal 10.0 g/dl
High Normal 18.0 g/dl
Linearity 20.0 g/dl

MANUAL PROCEDURE

PIPETTE INTO TEST TUBE
  BLANK STD SAMPLE
Sample - - 20 µl
Standard - 20 µl -
Reagent 5000 µl 5000 µl 5000 µl

Mix well, Incubate at r.t. for 5 mins. Measure final
absorbance of the sample (Ac) and standard (As)
against the reagent blank. At 540 nm (520-550 nm)

CALCULATION AND LINEARITY
Ac/As x Conc. Std. = g/dl HEMOGLOBIN

The hemoglobin assay shows a linearity upto 20 g/dl.
If the hemoglobin values are greater than 20 g/dl, dilute 1:1
with deionised water. Multiply result by 2.
Lab Care Diagnostics (India) Pvt. Ltd.
16-A, 'A' Wing , 1st Floor Sita Estate Mahul RoadAziz Baug, Chembur Mumbai 400 074. India
Phone: 00 91 22 2554 2109/ 00 91 22 2554 1558 Fax: 00 91 22 2554 3541
E-mail : accucare@labcarediagnostics.com