Development and validation of high performance liquid chromatography method for analysis of Raloxifene HCl in rat plasma by liquid –liquid extraction
A new, simple and sensitive HPLC method was developed for the quantification of raloxifene in rat plasma. Samples were prepared by liquid- liquid extraction method together with diclofenac sodium as internal standard (IS) at 2 μg/ml concentration. Separation of raloxifene from the treated samples wa...
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| Main Authors: | , , |
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| Format: | Conference or Workshop Item |
| Language: | English |
| Published: |
Kulliyyah of Pharmacy, International Islamic University Malaysia
2016
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/51874/13/51874-new.pdf http://irep.iium.edu.my/51874/ http://www.iium.edu.my/icip2016/ |
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| Summary: | A new, simple and sensitive HPLC method was developed for the quantification of raloxifene in rat plasma. Samples were prepared by liquid- liquid extraction method together with diclofenac sodium as internal standard (IS) at 2 μg/ml concentration. Separation of raloxifene from the treated samples was performed in a thermo-fisher syncronis RP- C18 column (150mm x 4.6 mm, 5μm particle size) at room temperature with a mobile phase containing acetonitrile and phosphate buffer of pH 6.1 (34:66 v/v) at a flow rate of 1 min/ml and ultraviolet detection at 285 nm. The developed method was validated for specificity, linearity, precision, accuracy, stability, sensitivity and recovery. Good linearity was achieved in the range of 125 ng/ml to 10 μg/ml (R2 = 0.9994). Precision and accuracy were within the acceptable limit of ICH guideline. Raloxifene was stable in quality control (QC) samples subjected to three freeze thaw cycles. QC Samples were also stable for 12 hours at room temperature, 3 months stored in deep freeze at -200C, and 12 hours after their processing (extraction followed by reconstitution with the mobile phase). The Overall recovery of raloxifene from the plasma samples were found to be within a range of 75.18% to 88.95%. The developed method was sensitive enough (LOQ: 125 ng/ml) that can be used to determine the plasma concentration of raloxifene in a pharmacokinetic study involving rats. |
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