2019J. Pl. Res. Vol. 17, No. 1, pp 118-124, 2019Journal of Plant Resources Vol.17, No. 1 Phytochemical, Microscopic and Standardization of Bergenia ciliata for Authentication Laxman Bhandari1*, Bal Bahadur Bista2 and Chetana Khanal1 1Natural Products Research Laboratory, Department of Plant Resources, Thapathali, Nepal 2Department of Plant Resources, Thapathali, Nepal *E-mail: [email protected] Abstract The present research was conducted to investigate the Bergenia ciliata (Haw.) Sternb. (Family- Saxifragaceae) for pharmacognostic study including macroscopical and microscopical observations, UV-fluorescence characters, preliminary phytochemical screening and thin layer chromatography of methanolic extracts. The macroscopy revealed that rhizome was barreled shaped, fibrous fracture surface, brown in color, solid rough texture, pleasant in odor and astringent in taste. While microscopy of rhizome showed typical dicot histological differentiation, no endodermis and pericycle seen. Vascular bundles are arranged in ring. Tanniferous cells are seen abundantly in a cortical region and pith. Rhizome powder has rosette crystals, starch grains, and vessels with scalariform thickenings. The physico-chemical parameters of rhizome powder showed loss on drying 7.04, total ash value 10.23, acid insoluble ash 0.11, water soluble ash 0.92, pH at 10% water solution is basic, water soluble extractive value 74.12, alcohol soluble extractive value 53.23 and extractive value of methanolic extracts 4.234 g. Phytochemicals reveled the presences of flavonoids, terpenoids, tannins, phenols and fatty acids and Rf value 0.74 in Co- TLC with standard bergenin in methanolic extracts. Fluorescence study indicates different colors like dark brown, red, brown black, creamy black, dark black, dark brownish black etc. that revealed the presence of different fluorescent chemical compounds. The rhizome of this plant has a high reputation in indigenous systems of medicine for various medicinal properties so the current research was done in Nepal and might be beneficial for public on accurate identification, in authentication intentional and unintentional adulteration in raw materials of the sample, quality assurance and will give a basis for its phyto-pharmacognostic characterization of the plants. Keywords: Co-TLC, Microscopic evaluation, Phytochemical screening, UV-Fluorescence Introduction 30 cm or more, turning bright red in autumn fringed with long bristle like hairs, leaf stalk short. Flowers Bergenia ciliata in sanskrit: Pashanbheda (Medicinal are white, pinkish or purple in spreading or dense Plants of Nepal, 2007) is a plant species in the clusters borne on a stout leafless stem. Fruits capsule, genus Bergenia is a small perennial rhizomatous 2 mm long (Ghimire, 2014; Pokhrel, 2014).The root creeping herb of family saxifragaceae found is bitter and acrid (Ahmad, 2018). The flowering throughout the temperate himalayans at an altitude season of this plant is March to April. (Khanal, 2018; of 900-3000m (Khan, 2017; Khanal, 2018). It was Ahmad, 2018).The root is used as a tonic in the long since known for its use in Ayurveda and has treatment of fevers, diarrhea and pulmonary been used as medicines for the treatment of different affections and juice is used to treat coughs and colds, human ailments (Khan, 2016). Bergenin, catechin, asthma and urinary problems (Rafi, 2017). It is also gallic acid, tannic acid, gallicin, catechin-7-O- considered helpful in relieving backache (Rajkumar, glucoside, limonene, afzelechin, arbutin, and - 2010). Besides its uses toxicity of this plant is sitosterol can be found in B.ciliata as a major reported (Ahmad, 2018). chemical constituents (Biradar, 2016; DPR, 2016). It is found on rocks ledges with stout rootstock The objectives of present study were microscopic (Ghimire, 2014). Leaves are simple, large with ovate study of plant materials and standardization of or rounded blade, 5 - 15 cm long but enlarging up to methanolic extract of these plants which is helpful 118 2019J. Pl. Res. Vol. 17, No. 1, pp 118-124, 2019Journal of Plant Resources Vol.17, No. 1 2019 Journal of Plant Resources Vol.17, No. 1 Phytochemical, Microscopic and Standardization of to identification of the plants parts and powder with freshly prepared acids, alkaline solution, Bergenia ciliata for Authentication material in public analysis. reagents and solvents and subjected to day light, short UV light 254nm and long UV light 366 nm for Laxman Bhandari1*, Bal Bahadur Bista2 and Chetana Khanal1 Materials and Methods visualization of color. 1Natural Products Research Laboratory, Department of Plant Resources, Thapathali, Nepal 2Department of Plant Resources, Thapathali, Nepal Plant material Soxhlet extraction *E-mail: [email protected] The plant material was collected in an appropriate 20 g each dried powder plant was extracted with Abstract stage of its growth from Pokhara, Kaski district near 250 ml methanol on Soxhlet Apparatus until the to Ghandruk Western part of Nepal in the month of solution of the sample in apparatus is colorless. The The present research was conducted to investigate the Bergenia ciliata (Haw.) Sternb. (Family- April. Plant material was authenticated through solution were evaporated in Rotatory Evaporator Saxifragaceae) for pharmacognostic study including macroscopical and microscopical under reduced pressure to get the viscous extract observations, UV-fluorescence characters, preliminary phytochemical screening and thin layer detail taxonomical study with compare to authentic chromatography of methanolic extracts. The macroscopy revealed that rhizome was barreled literatures and air- dried for study. Dried plant sample which was collected and dried for phytochemical shaped, fibrous fracture surface, brown in color, solid rough texture, pleasant in odor and was washed, cleaned, powdered in blender and screening and thin layer chromatography. astringent in taste. While microscopy of rhizome showed typical dicot histological differentiation, stored in air tight container for study of powder no endodermis and pericycle seen. Vascular bundles are arranged in ring. Tanniferous cells are microscopy, fluorescence, quality testing, extraction, Standardization for Quality Testing seen abundantly in a cortical region and pith. Rhizome powder has rosette crystals, starch grains, and vessels with scalariform thickenings. The physico-chemical parameters of rhizome powder thin layer chromatography profile and qualitative Total Ash: 2 g of sample was incarnated without showed loss on drying 7.04, total ash value 10.23, acid insoluble ash 0.11, water soluble ash 0.92, phytochemical screening. flaming at muffle furnace at 600ºc till residue is free pH at 10% water solution is basic, water soluble extractive value 74.12, alcohol soluble extractive from carbon (2 hrs.). The residue was cooled in value 53.23 and extractive value of methanolic extracts 4.234 g. Phytochemicals reveled the Macroscopic and organoleptic observations desiccators and amount of total ash is determined presences of flavonoids, terpenoids, tannins, phenols and fatty acids and Rf value 0.74 in Co- Z X TLC with standard bergenin in methanolic extracts. Fluorescence study indicates different colors Macroscopic evaluations were carried out through by weighing. Percentage of total ash = x100 like dark brown, red, brown black, creamy black, dark black, dark brownish black etc. that revealed appearance, texture and fracture surface in both fresh Y X the presence of different fluorescent chemical compounds. The rhizome of this plant has a high and dry states. Similarly organoleptic characters were where x = wt. of empty crucible, Y= wt. of crucible reputation in indigenous systems of medicine for various medicinal properties so the current identified through color, odor and taste of the sample with sample, Z= wt. of crucible with ash. research was done in Nepal and might be beneficial for public on accurate identification, in powder. authentication intentional and unintentional adulteration in raw materials of the sample, quality Acid Insoluble Ash: 25 mL of hydrochloric acid assurance and will give a basis for its phyto-pharmacognostic characterization of the plants. (about 70 g/L) was added to the crucible containing Histological evaluation the total ash, covered with watch glass and boiled Keywords: Co-TLC, Microscopic evaluation, Phytochemical screening, UV-Fluorescence Anatomy of fresh plant rhizome was studied through gently for 5 minutes. The watch glass was rinsed microscope by making permanent slide of transverse with 5 mL of hot water and added to crucible. The Introduction 30 cm or more, turning bright red in autumn fringed section of rhizome according standard method. The insoluble matter was collected on an ash less filter with long bristle like hairs, leaf stalk short. Flowers sections were observed through compound paper (Whatsmann 41) and washed with hot water Bergenia ciliata in sanskrit: Pashanbheda (Medicinal are white, pinkish or purple in spreading or dense microscope fitted with camera and photographs were until the filtrate was neutral. The filter-paper Plants of Nepal, 2007) is a plant species in the clusters borne on a stout leafless stem. Fruits capsule, taken. containing the insoluble matter was transferred to genus Bergenia is a small perennial rhizomatous 2 mm long (Ghimire, 2014; Pokhrel, 2014).The root the original crucible, ignited by gradually increasing creeping herb of family saxifragaceae found is bitter and acrid (Ahmad,