Chromatographic authentication of botanical origin: Herbaceous pollen profiling with HPLC, HPTLC and GC–MS analysis

This study describes a robust chromatographic authentication methodology forherbaceous pollen, employing gas chromatography–mass spectrometry (GC–MS),high-performance liquid chromatography (HPLC) and high-performance thin liquidchromatography (HPTLC) protocols. The comprehensive profiling of organiccompounds not only distinguishes between different botanical sources but alsoestablishes a reliable framework for quality control and assessment of herbaceouspollen authenticity. Traces of quercetin were detectable using HPTLC in Chaenomelesjaponica, and the composition of the mobile phase led to distinct phenolic acid tracksin the extracts of free phenolic compounds. In Lonicera nummulariifolia, prominentchlorogenic acid signal and traces of 3,4-dihydroxybenzoic acid were identified, alongwith the presence of vanillic, trans-ferulic, p-coumaric and p-hydroxybenzoic andsinapic as phenolic acid standards. The HPLC chromatogram identified six peaksrepresenting bioactive phenolic compounds such as gallic acid measuring 5.89± 0.56 mg g 1 , hydroxybenzoic acid 2.39 ± 0.78 mg g 1 and caffeic acid 2.83± 0.11 mg g 1 . The combined use of GC–MS, HPTLC and HPLC techniques providesa powerful and reliable means of authenticating the botanical origin of herbaceouspollen, offering valuable insights for quality control and ensuring the accuracy ofbotanical source identification