Bioanalytical Method Development, Validation and Stability Assessment of Xanthohumol in Rat Plasma

Harish, V; Almalki, WH; Alshehri, A; Alzahrani, A; Alzarea, SI; Kazmi, I; Gulati, M; Tewari, D; Chellappan, DK; Gupta, G; Dua, K; Singh, SK

Bioanalytical Method Development, Validation and Stability Assessment of Xanthohumol in Rat Plasma

Harish, V Almalki, WH Alshehri, A Alzahrani, A Alzarea, SI Kazmi, I Gulati, M Tewari, D Chellappan, DK Gupta, G Dua, K

Singh, SK

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Publisher:: MDPI AG
Publication Type:: Journal Article
Citation:: Molecules, 27, (20), pp. 7117-7117

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Field	Value	Language
dc.contributor.author	Harish, V
dc.contributor.author	Almalki, WH
dc.contributor.author	Alshehri, A
dc.contributor.author	Alzahrani, A
dc.contributor.author	Alzarea, SI
dc.contributor.author	Kazmi, I
dc.contributor.author	Gulati, M
dc.contributor.author	Tewari, D
dc.contributor.author	Chellappan, DK
dc.contributor.author	Gupta, G
dc.contributor.author	Dua, K https://orcid.org/0000-0002-7507-1159
dc.contributor.author	Singh, SK
dc.date.accessioned	2022-10-26T22:46:31Z
dc.date.available	2022-10-26T22:46:31Z
dc.identifier.citation	Molecules, 27, (20), pp. 7117-7117
dc.identifier.issn	1420-3049
dc.identifier.uri	http://hdl.handle.net/10453/162727
dc.description.abstract	<jats:p>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% v/v OPA (A): Methanol (B) was 15:85 v/v 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 R2 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.</jats:p>
dc.language	en
dc.publisher	MDPI AG
dc.relation.ispartof	Molecules
dc.relation.isbasedon	10.3390/molecules27207117
dc.rights	info:eu-repo/semantics/openAccess
dc.subject	0304 Medicinal and Biomolecular Chemistry, 0305 Organic Chemistry, 0307 Theoretical and Computational Chemistry
dc.subject.classification	Organic Chemistry
dc.title	Bioanalytical Method Development, Validation and Stability Assessment of Xanthohumol in Rat Plasma
dc.type	Journal Article
utslib.citation.volume	27
utslib.for	0304 Medicinal and Biomolecular Chemistry
utslib.for	0305 Organic Chemistry
utslib.for	0307 Theoretical and Computational Chemistry
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Health
pubs.organisational-group	/University of Technology Sydney/Faculty of Health/Graduate School of Health
pubs.organisational-group	/University of Technology Sydney/Faculty of Health/Graduate School of Health/GSH.Pharmacy
utslib.copyright.status	open_access	*
dc.date.updated	2022-10-26T22:46:29Z
pubs.issue	20
pubs.publication-status	Published online
pubs.volume	27
utslib.citation.issue	20

Abstract:

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% v/v OPA (A): Methanol (B) was 15:85 v/v 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 R2 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.

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http://hdl.handle.net/10453/162727