High Performance Liquid Chromatography (HPLC) Partially Purified Lantadene B Bioactive Pentacylic Triterpenoid from Lantana camara var. aculeate Leaves and It’s Trypanicidal Activity

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High Performance Liquid Chromatography (HPLC) Partially Purified Lantadene B Bioactive Pentacylic Triterpenoid from Lantana camara var. aculeate Leaves and It’s Trypanicidal Activity

August 24, 2021 Pharmaceutical 0

Lantana camara leaves were picked fresh, dried in the shade, then pounded. 3 x 40-ml MeCN was used to extract the obtained leaf powder (2g) of L. camara. Thin layer chromatography, column chromatography, preparative thin layer chromatography, and fractional crystallisation were used to purify the extract. After being dissolved in HPLC grade methanol, pooled fractions that contained bio constituents of latandene B as revealed on thin layer chromatography were taken for HPLC analysis. For HPLC analysis, aliquots (20 l) were collected. Waters (USA) provided the HPLC system, which included a 510 pump, a Rheodyne injector with a 20-l loop, a 490E multichannel detector, and a Millennium 2010 dataprocessor. The wavelength of the detector was set at 210 nm. The content of purified and spectroscopically determined lantadene B in young and matured leaves of L. camara was 347.0 3.0 and 522.3 37.1, respectively. On Vero cells cultured in Dulbecco’s Modified Eagle Medium (DMEM) and supplemented with foetal calf serum (FCS) 20-40% at suitable conditions, identified Lantadene B was evaluated for trypanocidal activity against Trypanosoma evansi. On Vero cells, an in vitro cytotoxicity test of lantadene B at doses (1.56-100 g ml-1) was performed without FCS. Trypanocidal activity in vitro ranged from immobilisation, reduction, and killing of trypanosomes in ELISA plate wells. At 250 g ml-1 pure Lantadene B, the average mean trypanosome count was reduced dramatically (from 40.0.0 to 1.6670.33). At 500 g ml-1 of test lantadene B, trypanosomes were completely killed (40.0.0 to 0.000.00) after 8 hours of incubation, which was statistically equivalent to diminazine aceturate (50 g ml-1) after 4 hours. With a significant difference (P 0.05 to 0. 01), trypanosome counts dropped in a concentration and time-dependent manner. Purified lantadene B and the standard drug diminazine aceturate were cytotoxic to Vero cells in an in vitro cytotoxicity assay at all doses except 6.25-1.56 g ml-1. At the same concentration, lantadene B and diminazine aceturate were cytotoxic to Vero cells. Higher anti-trypanosomal activity was attributed to lantadene B, which could be a future candidate chemical for a novel trypanocide.

Author (S) Details

Shaba Peter
Division of Medicine, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh (243 122), India.

Sharma Om Prakash
Formerly, Indian Veterinary Research Institute, Regional Station, Palampur Himachal Pradesh, (170), India.

Pandy Nitish Nerender
Division of Medicine, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh (243 122), India.

Sahab Dey
Division of Medicine, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh (243 122), India.

Jammi Rao
Formerly, Division of Parasitology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh (243 122), India.

Singh Raj Kumar
Indian Veterinary Research Institute, Regional Station, Mukteswar, Uttranchal, (263 138), India.

Bhanuprakash Veerakyathappa
Indian Veterinary Research Institute, Regional Station, Mukteswar, Uttranchal, (263 138), India.

Chaudary Pauload
Division of Bacteriology and Biotechnology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh (243 122), India.

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