Bioanalytical Method Development, Validation and Stability Assessment of Xanthohumol in Rat Plasma
Xanthohumol (XH) a prenylated chalcone has diverse therapeutic effects against various diseases. In the present study, a bioanalytical method was developed for XH in rat plasma using reverse phase high performance liquid chromatography. The validation of the method was performed as per ICH M10 guide...
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MDPI AG
2022-10-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/27/20/7117 |
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| author | Vancha Harish Waleed Hassan Almalki Ahmed Alshehri Abdulaziz Alzahrani Sami I. Alzarea Imran Kazmi Monica Gulati Devesh Tewari Dinesh Kumar Chellappan Gaurav Gupta Kamal Dua Sachin Kumar Singh |
| author_facet | Vancha Harish Waleed Hassan Almalki Ahmed Alshehri Abdulaziz Alzahrani Sami I. Alzarea Imran Kazmi Monica Gulati Devesh Tewari Dinesh Kumar Chellappan Gaurav Gupta Kamal Dua Sachin Kumar Singh |
| author_sort | Vancha Harish |
| collection | DOAJ |
| description | Xanthohumol (XH) a prenylated chalcone has diverse therapeutic effects against various diseases. In the present study, a bioanalytical method was developed for XH in rat plasma using reverse phase high performance liquid chromatography. The validation of the method was performed as per ICH M10 guidelines using curcumin as an internal standard. The Isocratic elution method was used with a run time of 10 min, wherein the mobile phase ratio 0.1% <i>v</i>/<i>v</i> OPA (A): Methanol (B) was 15:85 <i>v</i>/<i>v</i> at flow rate 0.8 mL/min and injection volume of 20 µL. The chromatograms of XH and curcumin was recorded at a wavelength of 370 nm. The retention time for XH and curcumin was 7.4 and 5.8 min, respectively. The spiked XH from plasma was extracted by the protein precipitation method. The developed method was linear with R<sup>2</sup> value of 0.9996 over a concentration range of 50–250 ng/mL along with LLOQ. The results of all the validation parameters are found to be within the accepted limits with %RSD value less than 2 and the percentage recovery was found to be greater than 95%. Based on the %RSD and percentage recovery results it was confirmed that the method was precise and accurate among the study replicates. LOD and LOQ values in plasma samples were found to be 8.49 ng/mL and 25.73 ng/mL, respectively. The stability studies like freeze thaw, short term and long-term stability studies were also performed, %RSD and percentage recovery of the XH from plasma samples were within the acceptable limits. Therefore, the developed bioanalytical method can be used effectively for estimation of XH in plasma samples. |
| first_indexed | 2024-03-09T19:41:46Z |
| format | Article |
| id | doaj.art-c87f0488a86c480aa4e11e486f7beaa8 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T19:41:46Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-c87f0488a86c480aa4e11e486f7beaa82023-11-24T01:37:43ZengMDPI AGMolecules1420-30492022-10-012720711710.3390/molecules27207117Bioanalytical Method Development, Validation and Stability Assessment of Xanthohumol in Rat PlasmaVancha Harish0Waleed Hassan Almalki1Ahmed Alshehri2Abdulaziz Alzahrani3Sami I. Alzarea4Imran Kazmi5Monica Gulati6Devesh Tewari7Dinesh Kumar Chellappan8Gaurav Gupta9Kamal Dua10Sachin Kumar Singh11School of Pharmaceutical Sciences, Lovely Professional University, Jalandhar-Delhi G.T Road, Phagwara 144411, Punjab, IndiaDepartment of Pharmacology, Umm Al-Qura College of Pharmacy, Umm Al-Qura University, Makkah 21955, Saudi ArabiaDepartment of Pharmacology & Toxicology, Faculty of Pharmacy, Northern Border University, Rafha 91911, Saudi ArabiaPharmaceuticals Chemistry Department, Faculty of Clinical Pharmacy, Al Baha University, Al Baha 65779, Saudi ArabiaDepartment of Pharmacology, College of Pharmacy, Jouf University, Sakaka 72341, Saudi ArabiaDepartment of Biochemistry, Faculty of Science, King Abdulaziz University, Jeddah 21589, Saudi ArabiaSchool of Pharmaceutical Sciences, Lovely Professional University, Jalandhar-Delhi G.T Road, Phagwara 144411, Punjab, IndiaDepartment of Pharmacognosy and Phytochemistry, School of Pharmaceutical Sciences, Delhi Pharmaceutical Sciences and Research University, New Delhi 110017, Delhi, IndiaSchool of Pharmacy, International Medical University, Bukit Jalil, Kuala Lumpur 57000, MalaysiaSchool of Pharmacy, Suresh Gyan Vihar University, Mahal Road, Jagatpura, Jaipur 302017, Rajasthan, IndiaFaculty of Health, Australian Research Centre in Complementary & Integrative Medicine, University of Technology Sydney, Ultimo, NSW 2007, AustraliaSchool of Pharmaceutical Sciences, Lovely Professional University, Jalandhar-Delhi G.T Road, Phagwara 144411, Punjab, IndiaXanthohumol (XH) a prenylated chalcone has diverse therapeutic effects against various diseases. In the present study, a bioanalytical method was developed for XH in rat plasma using reverse phase high performance liquid chromatography. The validation of the method was performed as per ICH M10 guidelines using curcumin as an internal standard. The Isocratic elution method was used with a run time of 10 min, wherein the mobile phase ratio 0.1% <i>v</i>/<i>v</i> OPA (A): Methanol (B) was 15:85 <i>v</i>/<i>v</i> at flow rate 0.8 mL/min and injection volume of 20 µL. The chromatograms of XH and curcumin was recorded at a wavelength of 370 nm. The retention time for XH and curcumin was 7.4 and 5.8 min, respectively. The spiked XH from plasma was extracted by the protein precipitation method. The developed method was linear with R<sup>2</sup> value of 0.9996 over a concentration range of 50–250 ng/mL along with LLOQ. The results of all the validation parameters are found to be within the accepted limits with %RSD value less than 2 and the percentage recovery was found to be greater than 95%. Based on the %RSD and percentage recovery results it was confirmed that the method was precise and accurate among the study replicates. LOD and LOQ values in plasma samples were found to be 8.49 ng/mL and 25.73 ng/mL, respectively. The stability studies like freeze thaw, short term and long-term stability studies were also performed, %RSD and percentage recovery of the XH from plasma samples were within the acceptable limits. Therefore, the developed bioanalytical method can be used effectively for estimation of XH in plasma samples.https://www.mdpi.com/1420-3049/27/20/7117xanthohumolrat-plasmabioanalytical methodRP-HPLCstability |
| spellingShingle | Vancha Harish Waleed Hassan Almalki Ahmed Alshehri Abdulaziz Alzahrani Sami I. Alzarea Imran Kazmi Monica Gulati Devesh Tewari Dinesh Kumar Chellappan Gaurav Gupta Kamal Dua Sachin Kumar Singh Bioanalytical Method Development, Validation and Stability Assessment of Xanthohumol in Rat Plasma Molecules xanthohumol rat-plasma bioanalytical method RP-HPLC stability |
| title | Bioanalytical Method Development, Validation and Stability Assessment of Xanthohumol in Rat Plasma |
| title_full | Bioanalytical Method Development, Validation and Stability Assessment of Xanthohumol in Rat Plasma |
| title_fullStr | Bioanalytical Method Development, Validation and Stability Assessment of Xanthohumol in Rat Plasma |
| title_full_unstemmed | Bioanalytical Method Development, Validation and Stability Assessment of Xanthohumol in Rat Plasma |
| title_short | Bioanalytical Method Development, Validation and Stability Assessment of Xanthohumol in Rat Plasma |
| title_sort | bioanalytical method development validation and stability assessment of xanthohumol in rat plasma |
| topic | xanthohumol rat-plasma bioanalytical method RP-HPLC stability |
| url | https://www.mdpi.com/1420-3049/27/20/7117 |
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