.

XXX County.
Standard Operating Procedure. SOP No: LHAEM XXX
Version: 1.
Supercedes: N/A.
Effective Date: XXX

Title: Haemoglobin Level Estimation using Diaspect Machine.

NAME. SIGNATURE. DATE.

PREPARED BY.

REVIEWED BY

APPROVAL
BY
FACILITY:____________________

1.0 PURPOSE / INTRODUCTION

1.1 Haemoglobin is measured to detect anaemia, its severity and to monitor an anaemic
patient’s response to treatment. Monitoring the haemoglobin level is also required
when patients with HIV disease are being treated with drugs such as AZT. The test
is also performed to check the haemoglobin level of a blood donor prior to donating
blood. Haemoglobin values are expressed in grams per litre (g/l) or grams per
decilitre (g/dl).

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1.2 This SOP is meant to give guidelines in ensuring that haemoglobin level estimation
using diaspect hb meter is done in accordance to the required laboratory standard to
give precise and accurate results.

2.0 SCOPE / RESPONSIBILITY

2.1 This SOP applies to all laboratory technologists/technicians involved in the assay.
2.2 It is the responsibility of the designated Quality Assurance and Quality Control
(QC/QA) personnel, Laboratory Supervisors, SCMLTs, and CMLT to ensure that
the current SOP is available to the staff and the procedure is followed as
documented.

3.0 SAFETY/RISK ASSESSMENT

3.1 Wear personal protective equipment. Handle all samples as potential biohazards.

4.0 DEFINITIONS
4.1 EDTA - Ethylene Diamine Tetra-acetic Acid
4.2 PPE - Personal Protective Equipment.
4.3 QA - Quality Assurance.
4.4 SOP - Standard Operating Procedure.

5.0 SPECIMEN
5.1 EDTA anticoagulated blood sample.
5.2 Capillary blood or Whole blood can also be used directly from a finger prick.

6.0 EQUIPMENT/ MATERIALS/ REAGENTS

6.1 Equipment.
6.1.1 DIASPECT HB machine/meter.
6.1.1.1 Operating Environment Temperature range 18–30 ºC with an upper
humidity limit of 90% non-condensing.
6.2 Materials.
6.2.1 Disposal containers and bucket.
6.2.2 Sterile lancet.
6.2.3 70% methanol or alcohol swabs.
6.2.4 Cotton wool or gauze.

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6.2.5 PPEs e.g. Gloves and a Lab coat.

6.3 Reagent.
6.3.1 Diaspect microcuvettes.

7.0 METHODOLOGY
7.1 Principle.
7.1.1 It is a non-dilution technique where blood is collected directly into a
specifically designed single-use microcuvette which is internally coated
with reagent (sodium deoxycholate) to lyze the blood. The haemoglobin
reacts with sodium nitrite forming methaemoglobin and sodium azide to
give azidemethaemoglobin which is measured by the Hemocue 301 meter at
specific wavelengths to compensate for any turbidity in the sample, e.g.
caused by high WBC count or raised lipids. There is no interference from
biliribin.
7.2 Procedure.
7.2.1 Confirm that the information on the request form is correct and properly
identify the patient/specimen. The collection of capillary blood and venous
blood are described in SOPs LGEN 0020 and 0021.
7.2.2 Assemble the microcuvette and other materials required to ensure easy
access and confirm tests requested.
7.2.3 Wear the appropriate personal protective equipment (PPE).
7.2.4 Pull out the microcuvette holder of the Hemocue 301 meter to its loading
position.
7.2.5 Press and hold down the left button of the meter until the display is
activated. The meter automatically carries out a performance check. After
10 seconds the meter will show three flashing dashes, indicating it is ready
for use.
7.2.6 Fill a microcuvette in one continuous process with capillary blood or well
mixed venous blood. Do not refill. Should air bubbles form, another
microcuvette must be used.
7.2.7 Wipe off excess blood from the outside of the microcuvette tip using a dry
cotton wool, making sure no blood is withdrawn.
7.2.8 Within 10 minutes of filling the microcuvette, Place the microcuvette in the
microcuvette holder and push in the holder to its measuring position.

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7.2.9 After 15–60 seconds, the haemoglobin value will be displayed (it will
remain displayed for as long as the holder is left in its measuring position).
7.2.10 Open the microcuvette holder and discard the microcuvette in sharps box.
7.3 Quality Control.
7.3.1 The meter is calibrated by the manufacturer. A performance check (self-
test) is inbuilt. Every time the meter is turned on and every 2 hours during
operation a performance check is carried out.
7.3.2 The chemicals in the microcuvettes are hygroscopic thus it is important to
protect the microcuvettes from heat and humidity.
7.3.3 The container in which the microcuvettes are stored must be kept tightly
capped.
7.4 Results.
7.4.1 Report results as displayed on the meter screen.
7.4.2 Example: “Hb Level - 10g/dl”
7.4.3 Indicate the date and name of technical staff reporting.
7.5 Limitations.
7.5.1 Not applicable.

8.0 APPENDICES
8.1 Not Applicable.

9.0 REFERENCES
9.1 AMREF (2008), Standard operating procedures for essential laboratory tests,
AMREF – MOH publication.
9.2 Monica Cheesbrough (2006), District laboratory practice in tropical countries,
Cambridge University press, Part 2.

10.0 DOCUMENT CHANGE HISTORY

1.1 Version Table:
Version 1:. Dated: SOP No. No. Pages:

Version 2: Dated: SOP No.: No. Pages:

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Version 3: Dated: SOP No.: No. Pages:

1.2 SOP Review Log.

Date of Changes made. Name of reviewer. Signature.
review.

2.0 SOP AWARENESS LOG.

I, the under named, have read and understand the contents of this SOP. I agree to
contact my supervisor/ designee if I have any query.

NO. DATE NAME SIGNATURE

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