SIMPLE METHOD FOR OPTIMIZATION OF HEPARIN AND ENOXA PARIN DETECTION USING AGAROSE GEL ELECTROPHORESIS
By: Almeman, Ahmad Abdulrahman
.
Publisher: M P Innovare Academic Sciences Pvt Ltd 2019Edition: Vol.1(4).Description: 12-15p.Subject(s): PHARMACEUTICSOnline resources: Click here
In:
International journal of pharmacy and pharmaceutical scienceSummary: Objective
:
A simple gel electrophoresis method for low-molecul
ar-weight heparins (LMWH) is required for use in a
variety of laboratories to allow further
identification and purification. This study aimed t
o optimize the detection of heparin and enoxaparin
(low-molecular-weight heparin by gel electrophoresi
s.
Methods
:
Several gel electrophoresis conditions were tested
to optimize the detection of enoxaparin by using a
simple method with a modified
Volpi’s approach. Multiple gel thicknesses, voltage
settings, and enoxaparin concentrations were teste
d in the optimization procedure. Enoxaparin
was purchased from a local supplier as pre-filled p
harmaceutical injections. Highly purified 0.5% and
1.0% agarose gels were prepared and a series
of enoxaparin concentrations was added to both gels
for comparison and optimization. The 0.2% toluidin
e blue stain was prepared by the addition
of 1 ml in an ethanol-water-acetic acid mixture (50
:49:1; v/v/w). The staining process comprised two s
teps: first, toluidine blue was added for 30
min and destained overnight in the solvent mixture.
Subsequently, the following morning, the second st
ep was conducted, in which the gel was
restained for 30 min with the same concentration of
toluidine blue. We continued to stain the gel unti
l the bands were visible.
Results:
The gel electrophoresis results showed that cleares
t and sharpest bands were obtained using 65–75 mAh
and 85 V settings. At 95 mAh, the
bands were slightly washed out.
Conclusion:
This study successfully facilitated the detection o
f enoxaparin, a LMWH, and heparin in the laboratory
by using simple tools and
techniques available in most laboratories.
| Item type | Current location | Call number | Status | Date due | Barcode | Item holds |
|---|---|---|---|---|---|---|
Articles Abstract Database
|
School of Pharmacy Archieval Section | Not for loan | 2020901 |
Objective
:
A simple gel electrophoresis method for low-molecul
ar-weight heparins (LMWH) is required for use in a
variety of laboratories to allow further
identification and purification. This study aimed t
o optimize the detection of heparin and enoxaparin
(low-molecular-weight heparin by gel electrophoresi
s.
Methods
:
Several gel electrophoresis conditions were tested
to optimize the detection of enoxaparin by using a
simple method with a modified
Volpi’s approach. Multiple gel thicknesses, voltage
settings, and enoxaparin concentrations were teste
d in the optimization procedure. Enoxaparin
was purchased from a local supplier as pre-filled p
harmaceutical injections. Highly purified 0.5% and
1.0% agarose gels were prepared and a series
of enoxaparin concentrations was added to both gels
for comparison and optimization. The 0.2% toluidin
e blue stain was prepared by the addition
of 1 ml in an ethanol-water-acetic acid mixture (50
:49:1; v/v/w). The staining process comprised two s
teps: first, toluidine blue was added for 30
min and destained overnight in the solvent mixture.
Subsequently, the following morning, the second st
ep was conducted, in which the gel was
restained for 30 min with the same concentration of
toluidine blue. We continued to stain the gel unti
l the bands were visible.
Results:
The gel electrophoresis results showed that cleares
t and sharpest bands were obtained using 65–75 mAh
and 85 V settings. At 95 mAh, the
bands were slightly washed out.
Conclusion:
This study successfully facilitated the detection o
f enoxaparin, a LMWH, and heparin in the laboratory
by using simple tools and
techniques available in most laboratories.
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