Seasonal Variation,Microscopic and Chromatographic Analysis of Leaves in Malus hupehensis:A Protocol

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Objective To establish a quality control protocol based on microscopic, TLC, and HPLC methods, and to verify the optimal harvesting time for the leaves of Malus hupehensis(LMH). Methods The LMH were pulverized into powder for microscopic identification or TLC and HPLC analysis after ultrasonic extraction with methanol. Seasonal variations of the phlorizin content and average leaf weight were determined by HPLC analysis and weighing up the leaves collected from May to October. Results Microscopic and macromorphologic characteristics have been described for the leaf identification. A qualitative TLC assay and a quantitative HPLC method have been established for the quality control of LMH. Phlorizin was selected as a reference marker, which resolved at Rf 0.53 in TLC assay and at 14.0 min in HPLC assay. The content of phlorizin decreased gradually from 17.0% in leaves collected in May to 7.5% in October. The average leaf weight reached the level of 0.6 g in August and maintained until its falling.Conclusion These methods are simple, selective, accurate, and reliable for the quality control of LMH. The period from late August to early September is suggested as the optimal harvesting time of the LMH. Objective To establish a quality control protocol based on microscopic, TLC, and HPLC methods, and to verify the optimal harvesting time for the leaves of Malus hupehensis (LMH). Methods The LMH were pulverized into powder for microscopic identification or TLC and HPLC analysis after Ultrasonic extraction with methanol. Seasonal qualitative of the phlorizin content and average leaf weight were determined by HPLC analysis and weighing up the leaves collected from May to October. Results Microscopic and macromorphologic characteristics have been described for the leaf identification. A qualitative TLC assay and a quantitative HPLC method have been established for the quality control of LMH. Phlorizin was selected as a reference marker, which resolved at Rf 0.53 in TLC assay and at 14.0 min in HPLC assay. The content of phlorizin decreased gradually from 17.0% in leaves collected in May to 7.5% in October. The average leaf weight reached the level of 0.6 g in August and maintained until it falling.Conclusion These methods are simple, selective, accurate, and reliable for the quality control of LMH. The period from late August to early September is as the optimal harvesting time of the LMH.
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