Development and validation of a high-performance thin-layer chromatography‒densitometric method and mass spectroscopy profiling for the determination of bioactive phytosterol from Manilkara zapota L. P. Royen leaves and correlating its antioxidant and antiinflammatory potential
{"title":"Development and validation of a high-performance thin-layer chromatography‒densitometric method and mass spectroscopy profiling for the determination of bioactive phytosterol from Manilkara zapota L. P. Royen leaves and correlating its antioxidant and antiinflammatory potential","authors":"","doi":"10.1007/s00764-023-00280-x","DOIUrl":null,"url":null,"abstract":"<h3>Abstract</h3> <p>A simple, reliable high-performance thin-layer chromatography (HPTLC)‒densitometry method was developed and validated for the quantification of β-sitosterol, a significant bioactive phytosterol, in <em>Manilkara zapota</em> L. P. Royen leaves. This method is combined with mass spectroscopy (MS) for relevant structural identification, responsible for their antioxidant and antiinflammatory therapeutic potential. The pet ether (PE), chloroform (CH), ethyl acetate (EA), and hydro-ethanolic (HE) leaf extracts were prepared using the cold maceration technique. These extracts were screened for preliminary qualitative tests, and based on the screening results, secondary metabolites such as flavonoids, phenols, steroids, terpenoids, and tannins were further quantified. Antioxidant potential was assessed using 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2-azino-bis-(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS), ferric reducing antioxidant power (FRAP), and nitric oxide (NO) free radical scavenging assays. antiinflammatory activity was evaluated through protein denaturation and human red blood cell (HRBC) membrane stability methods. Additionally, an HPTLC method was developed and validated for β-sitosterol quantification. Among all the extracts, the ethyl acetate leaf extract exhibited the highest steroidal content [50% inhibitory concentration (IC<sub>50</sub>) 2.92 ± 0.21 mg β-sitosterol equivalent (BSE)/g dry extract (DE)], flavonoid content [IC<sub>50</sub> 22.9 ± 0.25 mg quercetin equivalent (QE)/g DE], and phenolic content [IC<sub>50</sub> 103.8 ± 0.23 mg gallic acid equivalent (GAE)/g DE]. The ethyl acetate extract also demonstrated the highest efficacy against DPPH (IC<sub>50</sub> 16.35 ± 1.49), ABTS (IC<sub>50</sub> 17.52 ± 2.36), FRAP [41.32 ± 0.02 mg ascorbic acid (AA)/g crude extract], NO (IC<sub>50</sub> 17.13 ± 2.06), protein denaturation (IC<sub>50</sub> 31.75 ± 2.1), and HRBC membrane stability (IC<sub>50</sub> 24.8 ± 2.44). Correlation analysis further supported these findings. Furthermore, following the International Council for Harmonisation guidelines, an HPTLC method was developed, and 434.4 ng of β-sitosterol was quantified, with the highest content found in the ethyl acetate extract. Therefore, <em>M. zapota</em> leaf extract exhibits beneficial antioxidant and antiinflammatory effects due to its richness in phytosterols. This extract has the potential to be further explored as a therapeutic agent against inflammatory diseases.</p>","PeriodicalId":17616,"journal":{"name":"Jpc-journal of Planar Chromatography-modern Tlc","volume":null,"pages":null},"PeriodicalIF":1.1000,"publicationDate":"2024-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Jpc-journal of Planar Chromatography-modern Tlc","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1007/s00764-023-00280-x","RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
引用次数: 0
Abstract
A simple, reliable high-performance thin-layer chromatography (HPTLC)‒densitometry method was developed and validated for the quantification of β-sitosterol, a significant bioactive phytosterol, in Manilkara zapota L. P. Royen leaves. This method is combined with mass spectroscopy (MS) for relevant structural identification, responsible for their antioxidant and antiinflammatory therapeutic potential. The pet ether (PE), chloroform (CH), ethyl acetate (EA), and hydro-ethanolic (HE) leaf extracts were prepared using the cold maceration technique. These extracts were screened for preliminary qualitative tests, and based on the screening results, secondary metabolites such as flavonoids, phenols, steroids, terpenoids, and tannins were further quantified. Antioxidant potential was assessed using 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2-azino-bis-(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS), ferric reducing antioxidant power (FRAP), and nitric oxide (NO) free radical scavenging assays. antiinflammatory activity was evaluated through protein denaturation and human red blood cell (HRBC) membrane stability methods. Additionally, an HPTLC method was developed and validated for β-sitosterol quantification. Among all the extracts, the ethyl acetate leaf extract exhibited the highest steroidal content [50% inhibitory concentration (IC50) 2.92 ± 0.21 mg β-sitosterol equivalent (BSE)/g dry extract (DE)], flavonoid content [IC50 22.9 ± 0.25 mg quercetin equivalent (QE)/g DE], and phenolic content [IC50 103.8 ± 0.23 mg gallic acid equivalent (GAE)/g DE]. The ethyl acetate extract also demonstrated the highest efficacy against DPPH (IC50 16.35 ± 1.49), ABTS (IC50 17.52 ± 2.36), FRAP [41.32 ± 0.02 mg ascorbic acid (AA)/g crude extract], NO (IC50 17.13 ± 2.06), protein denaturation (IC50 31.75 ± 2.1), and HRBC membrane stability (IC50 24.8 ± 2.44). Correlation analysis further supported these findings. Furthermore, following the International Council for Harmonisation guidelines, an HPTLC method was developed, and 434.4 ng of β-sitosterol was quantified, with the highest content found in the ethyl acetate extract. Therefore, M. zapota leaf extract exhibits beneficial antioxidant and antiinflammatory effects due to its richness in phytosterols. This extract has the potential to be further explored as a therapeutic agent against inflammatory diseases.
期刊介绍：
JPC - Journal of Planar Chromatography - Modern TLC is an international journal devoted exclusively to the publication of research papers on analytical and preparative planar chromatography. The journal covers all fields of planar chromatography, on all kinds of stationary phase (paper, layer, gel) and with various modes of migration of the mobile phase (capillary action or forced flow).
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
