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Abstract
Toddalia asiatica Linn. is an important medicinal plant belonging to the family Rutaceae. The plant is well known for its antimalarial activity, which has been attributed to the presence of benzophenanthridine alkaloid nitidine in the roots of plants. A simple, rapid, sensitive, accurate, repeatable and robust HPTLC method has been developed and validated for the quantitative determination of nitidine in the dried roots and plant tissue culture extracts of T. asiatica. Nitidine was estimated at 332 nm by densitometry using Silica gel 60 F254 as stationary phase and chloroform:methanol (7:1, v/v), and as mobile phase. Linearity was observed in the concentration range of 25 -200 ng/spot for nitidine. The limit of detection and limit of quantitation were found to be 0.026 and 0.086 ng/spot respectively for nitidine. Developed method was validated according to the ICH guidelines with respect to precision, accuracy, specificity and robustness. The technique has been applied for the first time for the estimation of nitidine in roots and plant tissue culture extracts of T. asiatica. Statistical analysis data indicate the accuracy and reliability of the method.
Highlights
Toddalia asiatica Linn. (Rutaceae), commonly known as Lopez root, forest pepper and wild orange tree, is an important medicinal plant and has been used in folklore use in India and China from the 18th century
Various extracts of root bark were studied for in vitro antimalarial activity against Chloroquine-susceptible strains (K67, K39, M24, UPA, SL/D6, HB3 etc.) and Chloroquine-resistant strains (ItD12, FCR3, FCB etc.) of Plasmodium falciparum and compared with nitidine
Literature survey reveals that there are no reports on HPTLC determination of nitidine from the roots of T. asiatica
Summary
Toddalia asiatica Linn. (Rutaceae), commonly known as Lopez root, forest pepper and wild orange tree, is an important medicinal plant and has been used in folklore use in India and China from the 18th century. Various extracts of root bark were studied for in vitro antimalarial activity against Chloroquine-susceptible strains (K67, K39, M24, UPA, SL/D6, HB3 etc.) and Chloroquine-resistant strains (ItD12, FCR3, FCB etc.) of Plasmodium falciparum and compared with nitidine. Bioassay-directed fractionation of T. asiatica extract was studied and found that nitidine inhibited human lymphoblastoid cell killing by HIV-1 in vitro XTTbased anti-HIV assay [5,6]. HPTLC method is used for quantification of phytochemicals [10,11] and qualitative analysis of plant extracts [12], and for quality control of raw materials and standardization of polyherbal formulations [13,14]. Literature survey reveals that there are no reports on HPTLC determination of nitidine from the roots of T. asiatica. The present work illustrates the densitometric HPTLC method establishment and validation for quantitation of nitidine in roots and tissue culture extracts of T. asiatica
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