Decreased blood levels are associated with various anaemias. Increased levels are observed In patients with polycythemia or dehydration.
The brownish coloured cyanmethaemoglobin Is the product of almost all forms of haemoglobin found In blood except HbS and can be measured at 546 nM.
Cat. No. A 565
1.Drabkin's Reagent 1000 mL
Potassium Ferricyanide 0.60 mMol/L
Sodium Cyanide 0.77 mMol/L
Phosphate Buffer 1.00 mMol/L
Contains preservatives and stabilisers.
2.Cyanmethaemoglobin Standard 10 mL
Haemoglobin 15.06 Gms/dL
Equivalent to 15.06 Gms/dL in assay condition.(Contains buffers, stabilisers and preservatives).
Preparation of Working Reagent
All Reagents are ready to use.
Drabkin's Reagent is stable at least 18 months at 20-35°C, away from light. The standard is stable at 2-8°C for 18 months.
Fresh blood from finger prick is preferred. Anticoagulated whole blood can be used. Common anticoagulants such as Oxalate, EDTA or Heparin have no effect on this assay. (All samples should be handled as potential infective agents as no laboratory methods make conclusive findings for its safety. Therefore, adequate protective laboratory measures should
be taken while handling such materials).
||25°C to 35°C
||546 nM (530-550 nM)
|Cuvette Path Length
|Max. limit of Blank Reagent
|Final Colour Stability
| Pipette into 2 Test Tubes
| Drabkins Reagent No.1
| Sample (Fresh Blood)
2.Mix well. Incubate at RT for 5 minutes.
3.Read at 546 nM (530-550 nM) or GREEN filter against Blank. The final colour is stable for 30 minutes.
MICRO METHOD FOR DISCRETE ANALYSERS
Drabkin's Reagent................................ 2.5 mL
Sample.................................................... 0.01 mL
Mix well. Incubate at RT for 5 minutes. Read O.D. at 546 nM (530 - 550 nM)
against Blank. The final colour is stable for 30 minutes.
Multiply results by factor or compute as given under result columns.
| Pipette into 3 Test tubes
| Drabkin's Reagent
| Cyanmethemoglobin Standard
| Read O.D. at 546nM (530- 550nM) or GREEN filter.
Corresponding Hb concentration
2. Draw a line, expressing Gms/dL on the X-axis and optical densities on the Y-axis of a graph. Extrapolate the line within the limit of the dynamic range of this methodology (20Gms/dL).
NOTE: Programme the analyser using system parameters. A specific programme data
sheet may be provided for each analyser upon request.
Haemoglobin In Gms/dL = OD Test x Factor
OR = ---------------- x 0.06 x 251
OR = ---------------- x 0.06 x 15.06
Compare the result from the Calibration graph.
Since there is no acceptable Indian values established beyond arbitration, the values given here are of European references.
||13.5 to 18.0
||12.0 to 16.0
||13.6 to 19.6
Children below 10 years may show slightly lower
As with all diagnostic methods, the final diagnosis should
not be made on the result of a single test as well as
laboratory diagnosis must be confirmed with clinical
This reagent system is highly interfered by PAP reagent system used in GOD - POD, Cholesterol Oxidase, GPO-PAP, Uricase - PAP etc. For values higher than 20 Gms/dL dilute blood sample 1 to 2 with 0.9% Sodium Chloride an perform the assay. Multiply the result by dilution factor i.e. by 2 for 1:1 dilution.