{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T14:15:01Z","timestamp":1791987301342,"version":"4.4.0"},"reference-count":51,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2020,8,17]],"date-time":"2020-08-17T00:00:00Z","timestamp":1597622400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001321","name":"National Research Foundation","doi-asserted-by":"publisher","award":["107805"],"award-info":[{"award-number":["107805"]}],"id":[{"id":"10.13039\/501100001321","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Phytochemical Analysis"],"published-print":{"date-parts":[[2021,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Introduction<\/jats:title><jats:p>The minor phenolic constituents of <jats:italic>Cyclopia pubescens<\/jats:italic> Eckl. &amp; Zeyh. are unknown and one dimensional (1D) liquid chromatography (LC) is unable to provide sufficient separation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methodology<\/jats:title><jats:p>A two\u2010dimensional (2D) LC method incorporating normal\u2010phasehigh performance countercurrent chromatography (NP\u2010HPCCC) in the first dimension (<jats:sup>1<\/jats:sup>D) and reversed\u2010phase ultra\u2010high\u2010performance liquid chromatography (RP\u2010UHPLC) as the second dimension (<jats:sup>2<\/jats:sup>D) was developed. The analytical HPCCC method was subsequently scaled up to semi\u2010preparative mode and fractions pooled based on phenolic sub\u2010groups. The phenolic compounds in selected fractions were subsequently isolated using RP\u2010HPLC on a C<jats:sub>18<\/jats:sub> column. Isolated compounds were identified by nuclear magnetic resonance (NMR) spectroscopy. The absolute configurations of compounds were determined by optical rotation and electronic circular dichroism spectra. Sugars were identified by gas chromatography\u2010mass spectrometry (GC\u2013MS) analysis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The comprehensive off\u2010line 2D CCC \u00d7 LC method gave a good spread of the phenolic compounds. Orthogonality calculated using both the convex hull and conditional entropy methods were 81%. High\u2010resolution mass spectrometric fragmentation spectra obtained from a quadrupole\u2010time\u2010of\u2010flight instrument and ultraviolet\u2010visible (UV\u2010vis) spectral data were used to (tentatively) identify 32 phenolic compounds from the analytical CCC fractions. Of the seven isolated compounds, (2<jats:italic>S<\/jats:italic>)\u20105\u2010<jats:italic>O<\/jats:italic>\u2010[\u03b1\u2010<jats:sc>l<\/jats:sc>\u2010rhamnopyranosyl\u2010(1 \u2192 2)\u2010\u03b2\u2010<jats:sc>d<\/jats:sc>\u2010glucopyranosyl]eriodictyol (<jats:bold>3<\/jats:bold>) and (2<jats:italic>S<\/jats:italic>)\u20105\u2010<jats:italic>O<\/jats:italic>\u2010[\u03b1\u2010<jats:sc>l<\/jats:sc>\u2010rhamnopyranosyl\u2010(1 \u2192 2)\u2010\u03b2\u2010<jats:sc>d<\/jats:sc>\u2010glucopyranosyl]\u20105,7,3\u2032,4\u2032\u2010tetrahydroxyflavan (<jats:bold>4<\/jats:bold>) were newly identified in all plants. The other isolated compounds were identified as (2<jats:italic>S<\/jats:italic>)\u20105\u2010<jats:italic>O<\/jats:italic>\u2010[\u03b1\u2010<jats:sc>l<\/jats:sc>\u2010rhamnopyranosyl\u2010(1 \u2192 2)\u2010\u03b2\u2010<jats:sc>d<\/jats:sc>\u2010glucopyranosyl]naringenin (<jats:bold>1<\/jats:bold>), <jats:italic>R<\/jats:italic>\u2010neo\u2010eriocitrin (<jats:bold>2<\/jats:bold>), 3\u2010<jats:italic>O<\/jats:italic>\u2010\u03b1\u2010<jats:sc>l<\/jats:sc>\u2010arabinopyranosyl\u20103,4\u2010dihydroxybenzoic acid (<jats:bold>5<\/jats:bold>), 4\u2010<jats:italic>O<\/jats:italic>\u2010\u03b2\u2010<jats:sc>d<\/jats:sc>\u2010glucopyranosyl\u2010<jats:italic>Z<\/jats:italic>\u20104\u2010hydroxycinnamic acid (<jats:bold>6<\/jats:bold>) and 4\u2010(4\u2032\u2010<jats:italic>O<\/jats:italic>\u2010\u03b2\u2010<jats:sc>d<\/jats:sc>\u2010glucopyranosyl\u20104\u2032\u2010hydroxy\u20103\u2032\u2010methoxyphenyl)\u20102\u2010butanone (<jats:bold>7<\/jats:bold>).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Among the 32 compounds (tentatively) identified, only six were previously identified in <jats:styled-content style=\"fixed-case\"><jats:italic>Cyclopia pubescens<\/jats:italic><\/jats:styled-content> using 1D LC. Most of the isolated compounds were also identified for the first time in <jats:italic>Cyclopia<\/jats:italic> spp., improving the knowledge of the minor phenolic compounds of this genus.<\/jats:p><\/jats:sec>","DOI":"10.1002\/pca.2981","type":"journal-article","created":{"date-parts":[[2020,8,28]],"date-time":"2020-08-28T01:37:35Z","timestamp":1598578655000},"page":"347-361","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Comprehensive off\u2010line CCC \u00d7 LC\u2010DAD\u2010MS separation of <i>Cyclopia pubescens<\/i> Eckl. &amp; Zeyh. phenolic compounds and structural elucidation of isolated compounds"],"prefix":"10.1002","volume":"32","author":[{"given":"Nico A.","family":"Walters","sequence":"first","affiliation":[{"name":"Plant Bioactives Group Post\u2010Harvest &amp; Agro\u2010Processing Technologies, Agricultural Research Council (ARC), Infruitec\u2010Nietvoorbij  Stellenbosch South 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