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The purpose of this paper is to validate the antimicrobial activity (both antibacterial and antifungal) of in vivo and in vitro ethanolic anthocyanin extracts of Clitoria ternatea L. (vivid blue flower butterfly-pea) and Dioscorea alata L. (purple yam) against selected bacteria (Bacillus subtilis, Staphylococcus aureus and Escherichia coli) and fungi (Fusarium sp., Aspergillus niger and Trichoderma sp.).

The freeze-dried samples (0.2 g) from in vivo vivid blue flowers of C. ternatea L. were extracted using 10 mL ethanol (produced ethanolic red extraction) and 10 mL distilled water (produced aqueous blue extraction) separately. Two-month-old in vitro callus samples (0.2 g) were only extracted using 10 mL ethanol. The anthocyanin extractions were separated with the addition (several times) of ethyl acetate and distilled water (1:2:3) to remove stilbenoids, chlorophyll, less polar flavonoids and other non-polar compounds. Furthermore, the antimicrobial properties were determined using agar diffusion technique. Three bacteria (B. subtilis, S. aureus and E. coli) and fungi (F. sp., A. niger and T. sp.) were streaked on bacteria agar and dextrose agar, respectively, using “hockey stick”. Then, the sterile paper discs (6 mm diameter) were pipetted with 20 µL of 1,010 CFU/mL chloramphenicol (as control for antibacterial) and carbendazim (as control for antifungal) in vivo and in vitro extracts. The plates were incubated at room temperature for 48 h, and the inhibition zones were measured.

Based on the results, both in vivo and in vitro ethanolic extracts from vivid blue flowers of C. ternatea L. showed the best antibacterial activity against the same bacteria (B. subtilis), 11 and 10 mm inhibition zones, respectively. However, different antifungal activity was detected in in vitro ethanolic callus extract (12 mm), which was against T. sp., contrary to in vivo ethanolic extract (10 mm), which was against F. sp.; antibacterial activity of D. alata L. was seen against the same bacteria (E. coli) with the highest inhibition zone for in vivo extract (8.8 mm), followed by in vitro extract (7.8 mm).

Anthocyanins are responsible for the water soluble and vacuolar, pink, red, purple and blue pigments present in coloured plant pigments. These pigments (pink, red, purple and blue) are of important agronomic value in many crops and ornamental plants. However, anthocyanins are not stable and are easy to degrade and fade whenever exposed to light.

Plant extracts containing bioactive agents with antimicrobial properties have been found to be useful in treating bacterial and fungal infections, as well as showed multiple antibiotic resistance.

Both in vivo and in vitro extracts from vivid blue flower petals (C. ternatea L.) and purple yam (D. alata L.) have important applications as natural antimicrobial (antibacterial and antifungal) agents in the coating industry, instead of natural pharmaceutical products.

This paper aims to study the effect of different organic solvents on the extraction of pigments present in callus cultures of E. cinerascens.

Attempts have been made to extract pigments from callus cultures through tissue culture system as an alternative replacement for conventional plant cultivation as tissue culture provides unlimited supplies of plant samples. Callus of E. cinerascens was induced from stem explant cultured in Murashige and Skoog medium supplemented with combination of 0.5 mg/L 6-benzylaminopurine and 0.5 mg/L α-naphthaleneacetic acid maintained under photoperiod of 16 h light and 8 h dark. Fresh samples of the callus were harvested and dissolved in various types and concentrations of solvents such as 100 per cent acetone, 80 per cent acetone, 95 per cent ethanol, 100 per cent methanol and 90 per cent methanol. Each of the mixtures was directly centrifuged to get clear supernatant containing pigments of interest. The pigments were detected and subsequently quantified via two simple techniques, ultraviolet-visible (UV-Vis) spectrophotometer and thin layer chromatography (TLC).

UV-Vis spectrophotometer detected two families of pigments present in the callus cultures, namely, carotenoids (carotene and xanthophyll) and tetrapyrroles (chlorophyll a and b). Pigment contents in various solvent extractions were estimated using spectroscopic quantification equations established. Through TLC, spots were seen on the plates, and Rf values of each spots were assessed to indicate the possible existence of carotenoids and tetrapyrroles.

This preliminary study offers significant finding for further advance research related on natural pigments extracted from E. cinerascens that would provide profits in the future applications, especially in food industry, medicine, agriculture, etc.