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Indian Journal of Community Medicine

Comparison of Hemoglobin Estimates from Filter Paper, Cyanmethemoglobin and HemoCue Methods

Author(s): P. Pathak, S. K. Kapoor, S.N. Dwivedi, P. Singh, U. Kapil

Vol. 29, No. 3 (2004-07 - 2004-09)


Objective: To compare the hemoglobin estimates obtained from Filter Paper Cyanmethemoglobin and HemoCue Method.

Study Design: A Hospital based study.

Setting: A hospital of a rural area in Haryana.

Participants: 334 adult patients attending a rural hospital, requiring hematological investigations.

Statistical Analysis: Means, Standard deviations.

Results: It was revealed that the HemoCue overestimated the hemoglobin values when compared to the Filter Paper Cyanmethemoglobin method by 1.8 g/dl.

Conclusion: The findings of the present study revealed that the HemoCue Method overestimated the hemoglobin values by 1.8 g when compared with the Filter Paper Cyanmethemoglobin Method.

Key words: Hemoglobin Estimation, Filter Cyanmethemoglobin, HemoCue Method


Iron Deficiency Anemia (IDA) is defined as a condition where the hemoglobin (Hb) concentratin of an individual is lower than the level considered normal for the person's age and sex. Laboratory tests are recommended to diagnose anemia and determine its severity1. There are several laboratory techniques for measuring hemoglobin concentration. The most widely used quantitative estimation of Hb is the Cyanmethemoglobin method2. For field studies, the scientists have recommended the Filter Paper Cyanmethemoglobin Method as it is simple and provides reliable results. The HemoCue method is also being used for Hb estimation in laboratory and in field conditions3,4. Concerns have been raised on the validity of the hemoglobin estimations obtained by the HemoCue3,5,6. The present study was undertaken with the objective to compare the hemoglobin estimation obtained by the Filter Paper Cyanmethemoglobin method routinely utilized in the research studies with the hemoglobin estimates of the HemoCue method.

Material and Methods

A hospital-based study, conducted in the months of May to July 2001. Adult patients attending a rural hospital, requiring hematological investigations, were enrolled. The informed consent was taken. Blood was collected by venous-puncture in EDTA vial (l mg/dl) from 334 subjects. The sample was mixed properly to avoid coagulation. For estimation of Hb by Filter Paper Cyanmethemoglobin method, 20 ml of blood was taken with the help of hemoglobin pipette on to a Whatman Filter Paper No. 1, utilizing the standard methodology1. The blood was allowed to dry on the filter paper. the portion of the filter paper with blood was cut and eluted in 5ml Drabkin's solution for 2 hours. The solution was put on vortex for complete elution of blood. The hemoglobin concentration was read at a wavelength of 540 nm utilizing a spectrophotometer. For estimation of Hb by HemoCue method, a drop of blood was pipetted onto a clean glass-slide. The micro-cuevette was applied to the blood, which filled, automatically by capillary action and subsequently, the hemoglobin estimaiton was done utilizing the recommended standard methodology3. It was ensured that the micro-cuevettes were taken for Hb estimaiton from tighly sealed containers and were within the expiry dates. The data collected was subjected to the standard statistical methods to obtain the results.


A total of 334 patients participated in the study. Comparison of hemoglobin levels obtained by Filter Paper Cyanmethemoglobin and HemoCue method are shown in Table I. It was observed that at various levels of the Hb, i.e. >7.0 g/dl, 7.0-< 10.0 g/dl, and 10.0 g/dl and more, the HemoCue Method over estimated the Hb as compared with the results of the Filter Paper Cyanmethemoglobin method. On application of the standard statistical method on the data, it was revealed that the HemoCue method over estimated the Hb values by 1.8g/dl.

Table I: Comarison of Hemoglobin Levels Obtained by Cyanmethemoglobin Filter Paper Method and HemoCue Method

Hemoglobin level (g/dl)
by Filter Paper
n Mean+SD* Mean+SD**
<7.0 45 5.5 + 1.2 6.6 ± 1.45
7.0-<10.0 178 8.9 ± 0.77 10.7 ± 1.23
10.0 and more 111 10.7 ± 0.62 12.8 ± 0.93
Total 334 9.0 ± 1.82 10.8 ± 2.23
* Filter Paper Cyanmethemoglobin Method ** HemoCue Method


The findings of the present study revealed that tthe HemoCue method over-estimated hemoglobin levels by a value of 1.8g/dl when compared to the Filter Paper Cyanmethemoglobin method. Other studies have also documented similar results3,5,6. In an earlier study, Hb levels when compared by the methods of Blood Cell Counter (BCC) and the HemoCue method revealed an over-estimation of the values by the HemoCue method. A correction factor of minus 0.5 g/dl was suggested to achieve comparable hemoglobin levels to the BCC5. Another study compared the Hb estimates obtained by the direct Cyanmethemoglobin, the Filter Paper Cyanmethemoglobin and the HemoCue methods. The study revealed an over-estimation of Hb levels by the HemoCue when compared to the direct and Filter Paper Cyanmethemoglobin methods by a factor of 0.18 and 0.45g/dl, respectively. This study documented that hot and humid climates, and duration of opening of the micro-cuevette box could affect the Hb estimates as estimated by HemoCue3. A recent study documented an overestimation of Hb values by 2.08 g/dl when compared to the standard Cyanmethemoglobin method6.


The findings of the present study revealed that the HemoCue Method overstimated the hemoglobin values by 1.8 g when compared with the Filter Paper Cyanmethemoglobin Method.


The authors would like to acknowledge the infrastructure facilities provided by the Director, All India Institute of Medical Sciences, New Delhi.


  1. DeMaeyer EM, Dallman P, Gurney JM, Hallberg L, Sood SK, Srikantia SG. Preventing and controlling iron deficiency anemia through primary health care. A guide for health administrators and programme managers. World Health Organization, Geneva 1989.
  2. Program for appropriate Technology in Health (Path). Anemia detection in Health Services Guideline for Program Managers. U.S. Agency for International Development 1996;37.
  3. Sari M, Pee de S, Martini E, Herman S, Sugiatmi, Bloem WM, Yip R. Estimating the prevalence of anemia: a comparison of three methods. Bulletin of the World Health Organisation 2001; 79:506-11.
  4. Vanzetti G. Statistical research methods. J. Lab. and Clin Med. 1966;67:116-7.
  5. Prakash S, Kapil U, Singh G, Dwivedi SN, Tandon M. Utility of HemoCue in estimation of hemoglobin against standard blood cell counter method. Journal of the Association of Physicians of India 1999;47:995-7.
  6. Mohanram M, Rao GVR, Gowrinath SJ. A comparative study of prevalence of anemia in women by Cyanmethemoglobin and HemoCue methods. Indian J Comm Med. 2002; 27:58-61.

Department of Human Nutrition,
Center for Community Medicine
Department of Biostatistics, All Indian Institute of Medical Sciences
Ansari Nagar, New Delhi, India

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