Validation of HPLC and Liquid-Liquid Extraction Methods for Warfarin Detection in Human Plasma and its Application to a Pharmacokinetics Study
A reversed-phase HPLC method to determine total plasma warfarin was developed and validated. Warfarin was extracted from human plasma using a two-step liquid-liquid extraction method. The residue was reconstituted with a phenylbutazone standard solution, which was used as the internal standard. The...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Academy of Sciences Malaysia
2019-06-01
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| Series: | ASM Science Journal |
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| Online Access: | https://doi.org/10.32802/asmscj.2019.215 |
| _version_ | 1828010318023884800 |
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| author | Yung An Chua Wan Zaidah Abdullah Zurkurnai Yusof Siew Hua Gan |
| author_facet | Yung An Chua Wan Zaidah Abdullah Zurkurnai Yusof Siew Hua Gan |
| author_sort | Yung An Chua |
| collection | DOAJ |
| description | A reversed-phase HPLC method to determine total plasma warfarin was developed and validated. Warfarin was extracted from human plasma using a two-step liquid-liquid extraction method. The residue was reconstituted with a phenylbutazone standard solution, which was used as the internal standard. The analytical column was a Purospher STAR RP-18e (4 x 4mm I.D., 5m particle size). The mobile phase consisted of acetonitrile: potassium dihydrogen orthophosphate buffer solution at pH 6.5 [30:70 (v/v)] with a flow rate of 1mL/min. Both warfarin and phenylbutazone were detected using a photodiode array detector. The lower limit of quantification was 100ng/mL, while the limit of detection was 20ng/mL. The linearity of the assay was good (r2=0.992) in the concentration range from 0.1 - 6.0µg/mL. The extraction recovery of warfarin was 93.53 ± 12.40%. Both the intraday and interday quality control assay for warfarin demonstrated good precision and accuracy, with all of the percentage coefficients of variation being less than 15%. Warfarin was stable in human plasma for up to three months of storage. The validated method was successfully applied to four human samples for a pharmacokinetics study. |
| first_indexed | 2024-04-10T08:52:09Z |
| format | Article |
| id | doaj.art-3bd168fdc84e4978af72c87f5fc15627 |
| institution | Directory Open Access Journal |
| issn | 1823-6782 |
| language | English |
| last_indexed | 2024-04-10T08:52:09Z |
| publishDate | 2019-06-01 |
| publisher | Academy of Sciences Malaysia |
| record_format | Article |
| series | ASM Science Journal |
| spelling | doaj.art-3bd168fdc84e4978af72c87f5fc156272023-02-22T03:11:21ZengAcademy of Sciences MalaysiaASM Science Journal1823-67822019-06-011211010.32802/asmscj.2019.215215Validation of HPLC and Liquid-Liquid Extraction Methods for Warfarin Detection in Human Plasma and its Application to a Pharmacokinetics StudyYung An Chua0Wan Zaidah Abdullah1Zurkurnai Yusof2Siew Hua Gan3Universiti Sains MalaysiaUniversiti Sains MalaysiaUniversiti Sains MalaysiaUniversiti Sains MalaysiaA reversed-phase HPLC method to determine total plasma warfarin was developed and validated. Warfarin was extracted from human plasma using a two-step liquid-liquid extraction method. The residue was reconstituted with a phenylbutazone standard solution, which was used as the internal standard. The analytical column was a Purospher STAR RP-18e (4 x 4mm I.D., 5m particle size). The mobile phase consisted of acetonitrile: potassium dihydrogen orthophosphate buffer solution at pH 6.5 [30:70 (v/v)] with a flow rate of 1mL/min. Both warfarin and phenylbutazone were detected using a photodiode array detector. The lower limit of quantification was 100ng/mL, while the limit of detection was 20ng/mL. The linearity of the assay was good (r2=0.992) in the concentration range from 0.1 - 6.0µg/mL. The extraction recovery of warfarin was 93.53 ± 12.40%. Both the intraday and interday quality control assay for warfarin demonstrated good precision and accuracy, with all of the percentage coefficients of variation being less than 15%. Warfarin was stable in human plasma for up to three months of storage. The validated method was successfully applied to four human samples for a pharmacokinetics study.https://doi.org/10.32802/asmscj.2019.215hplcliquid-liquid extractionmethod validationpharmacokineticswarfarin |
| spellingShingle | Yung An Chua Wan Zaidah Abdullah Zurkurnai Yusof Siew Hua Gan Validation of HPLC and Liquid-Liquid Extraction Methods for Warfarin Detection in Human Plasma and its Application to a Pharmacokinetics Study ASM Science Journal hplc liquid-liquid extraction method validation pharmacokinetics warfarin |
| title | Validation of HPLC and Liquid-Liquid Extraction Methods for Warfarin Detection in Human Plasma and its Application to a Pharmacokinetics Study |
| title_full | Validation of HPLC and Liquid-Liquid Extraction Methods for Warfarin Detection in Human Plasma and its Application to a Pharmacokinetics Study |
| title_fullStr | Validation of HPLC and Liquid-Liquid Extraction Methods for Warfarin Detection in Human Plasma and its Application to a Pharmacokinetics Study |
| title_full_unstemmed | Validation of HPLC and Liquid-Liquid Extraction Methods for Warfarin Detection in Human Plasma and its Application to a Pharmacokinetics Study |
| title_short | Validation of HPLC and Liquid-Liquid Extraction Methods for Warfarin Detection in Human Plasma and its Application to a Pharmacokinetics Study |
| title_sort | validation of hplc and liquid liquid extraction methods for warfarin detection in human plasma and its application to a pharmacokinetics study |
| topic | hplc liquid-liquid extraction method validation pharmacokinetics warfarin |
| url | https://doi.org/10.32802/asmscj.2019.215 |
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