Development and validation of bioanalytical method for the detection of gliclazide in human plasma
A robust High Performance Liquid Chromatography Diod Array Detection (HPLC- DAD) method was developed for the detection of gliclazide in human plasma. Liquid liquid extraction method was used as a sample preparation for HPLC analysis. The separation was carried out on Agilent Zorbax Reversed P...
| Main Authors: | , , , |
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| Format: | Conference or Workshop Item |
| Language: | English |
| Published: |
2016
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/55423/ http://irep.iium.edu.my/55423/1/Development%20and%20Validation%20of%20Bioanalytical%20Method%20for%20the%20Detection.pdf |
| Summary: | A robust High Performance Liquid Chromatography Diod Array Detection (HPLC- DAD) method
was developed for the detection of gliclazide in human plasma. Liquid liquid extraction method
was used as a sample preparation for HPLC analysis. The separation was carried out on Agilent
Zorbax Reversed Phase C-18 column (150 x 4.6 mm, 5µ m) with PDA detection at 236 nm. with
an isocratic elution (1 mL/min) at 30°C column temperature. The mobile phases used consisted
of acetonitrile: 0.3% triethylamine buffer (pH=3.0 adjusted with H3PO4 85%) 57:43 v/v. Then,
10 µL aliquot samples were injected onto chromatographic system. The results showed that the
retention time for the gliclazide and the internal standard, glibenclamide were 3.8 and 4.8
respectively. The method was linear over the range of 10 – 5000 ppb with R2 of 0.9971. The
recoveries of the developed methods for gliclazide were found to be between 84.9% and
104.0%, while for the glibenclamide were found to be between 100.0% and 112.0%. The
stability analysis showed that gliclazide is stable for at least 3 months but glibenclamide is
degraded for about 2 months when stored at -20 ?. For conclusion, the develop method is
suitable and reliable for the detection of gliclazide in human plasma. |
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