Assessment of the Total Flavonoid, Phenol, Alkaloid Content and Sun Protection Factor in Grewia abutilifolia Leaf Extract

A variety of secondary metabolites provide a valuable source of treatment for various pharmacological ailments. Grewia abutilifolia is a shrub found in the family Tiliaceae that is also investigated for its nutritional and medicinal benefits. Grewia is reported for its good antioxidant potential, which is applied to protect skin from UV rays. The main goal of this study is to determine the in-vitro sunscreen effect of Grewia abutilifolia based on phytochemical evidence of UV protection. Chemical treatments used to protect skin from the sun have several drawbacks, including numerous hazardous side effects on humans and the environment. The hydroalcoholic, ethyl acetate, and methanolic extracts were studied for their phytochemical, physicochemical, and sun protection activities. Total phenolic content, total alkaloid content, total flavonoid content, and SPF value were all highest in the methanolic leaf extract, with values of 99.20 0.12, 47.661.527, 96.02, and 14.50.01 for total phenolic content, total alkaloid content, total flavonoid content, and SPF value, respectively.

The Cultivation of Chelidonium majus L. Increased the Total Alkaloid Content and Cytotoxic Activity Compared with Those of Wild-Grown Plants

The effect of cultivation practises on both the phytochemical profile and biological activity of aqueous ethanol extracts of Chelidonium majus L. was studied. Extracts were prepared from aerial parts of the same plant population collected in the wild and grown under organic farming conditions. Both qualitative and quantitative analyses of alkaloids and flavonoid derivatives were performed by LC/MS methods, and the cytotoxicity of lyophilised extracts was studied in B16-F10, HepG2, and CaCo-2 cells. Coptisine was the dominant alkaloid of extracts prepared from wild-grown plants, whereas after cultivation, chelidonine was the most abundant alkaloid. The total alkaloid content was significantly increased by cultivation. Ten flavonol glycoconjugates were identified in C. majus extracts, and quantitative analysis did not reveal significant differences between extracts prepared from wild-grown and cultivated specimens. Treatment with C. majus extracts resulted in a dose-dependent increase in cytotoxicity in all three cell lines. The extracts prepared from cultivated specimens showed higher cytotoxicity than the extracts prepared from wild-grown plants. The strongest cytotoxic effect of cultivated C. majus was observed in B16-F10 cells (IC50 = 174.98 ± 1.12 µg/mL). Cultivation-induced differences in the phytochemical composition of C. majus extracts resulted in significant increases in the cytotoxic activities of the preparations.

Optimasi Penggunaan High Shear Mixer pada Pembuatan Fraksi Alkaloid dari Daun Awar-awar (Ficus septica) dengan Desain Faktorial

Awar-awar (Ficus septica) is an Indonesian anticancer plant that contains alkaloids as the active compound. The n-hexane insoluble fraction (FTLH) is alkaloid-containing fraction obtained from the ethanolic extract of F. septica leaves (EEDFS) by fractionation using n-hexane. High shear mixer (HSM) can be used to optimize the separation processes, including fractionation. The application of HSM in the fractionation of EEDFS is affected by fractionation duration, amount of solvent per gram extract, and strirring strength. Thus, these parameters must be optimized to obtain the optimum condition for the production of FTLH with the highest alkaloid content. This study aimed to optimize the production of FTLH using HSM with factorial designs. The single factor experimental design was employed to determine the influence of the individual variable on the alkaloid content and to define the optimum range value of each variable. A full factorial design was used to determine the presence of interaction among the factors and to determine the optimal fractionation condition. The results showed that all investigated factors independently affected alkaloid contents. The solvent volume of 14-18 mL per gram extract, the fractionation duration of 2.5-5.0 minutes, and the stirring strength of 3000 rpm are the optimal range value of each factor. The optimal conditions were solvent volume of 14.0-14.4 mL/gram extract, fractionation duration of 2.5-2.7 minutes, and strirring strength at 3000 rpm. At the optimal condition, the total alkaloid content in the FTLH reached 0.1466% or 1.3 times higher than the extract (0.1128%).

Pharmacognostical evaluation and anti-convulsant property of Annona reticulata Linn. (Annonaceae) root

Background The present investigation was aimed at establishing the pharmacognostical parameters and preliminary phytochemical screening of Annona reticulata Linn. (Annonaceae) root. Pharmacognostical studies and phytochemical evaluation of A. reticulata root were carried out by following standard procedures to provide information that help to identify the species. The species of Annonaceae family are documented to possess anticonvulsant property, while A. reticulata and its root are reported to be used as a folklore medicine to treat epilepsy. In this study, anti-convulsant activity of the root extract was further investigated in mouse model with seizures induced by pentylenetetrazole (PTZ; 60 mg/kg, i.p.). Diazepam (5 mg/kg) was used as a standard anti-convulsant drug. Prior to induction of seizures, ethanol extract (100, 200, and 400 mg/kg, p.o.,) and 0.1% sodium CMC were administered. Later, the onset, duration of convulsions along with recovery was recorded. Results A. reticulata can be recognized by its crown shape, leaves with lengthy petioles, fleshy petals and heart-shaped fruits. Roots are cylindrical with rootlets and have aromatic odour. Presence of oil globules and rhomboidal calcium oxalate crystals in cortex, sclerenchyma cells in cortex and secondary phloem, starch grains in cortex, secondary phloem and secondary xylem, and uni- to tri-seriate medullary rays are the key diagnostic characters of root. Preliminary phytochemical screening of extracts revealed the presence of phenolic compounds, tannins, alkaloids, carbohydrates, flavonoids, glycosides, saponins, proteins, fixed oil and fats. The total alkaloid content in the root was 0.524% w/w, while total flavonoid and total phenolic content in ethanol extract were 16.65 mg QE/g and 59.54 mg GAE/g, respectively. Based on the values obtained from anti-convulsant activity, it is evident that the ethanol extract offered significant protection against PTZ-induced convulsion in mice. Conclusion The current pharmacognostical study aids not only in identification of crude drug material but also in establishing the standardization parameters. Further, the findings of this study indicated that the ethanolic extract of Annona reticulata Linn root displayed significant anti-convulsant property and this property could be attributed to imperative bioactives like flavonoids, phenolic compounds, alkaloids, in addition to other secondary metabolites.

Improving large-scale biomass and total alkaloid production of Dendrobium nobile Lindl. using a temporary immersion system and MeJA

Background Dendrobium nobile Lindl. is an important pharmacopeial plant with medicinal and ornamental value. This study sought to provide technical means for the large-scale artificial production of total alkaloids in D. nobile. Seedlings were cultured in vitro in a temporary immersion bioreactor system (TIBS). The four tested immersion frequencies (min/h; 5/2, 5/4, 5/6, and 5/8) influenced the production of biomass and total alkaloid content. In addition, to compare the effects of different concentrations of phytohormone methyl jasmonate (MeJA) and treatment time on biomass and total alkaloid accumulation, MeJA was added to the TIBS medium after 50 days. Finally, the production of total alkaloids in a semi-solid system (SSS), TIBS, and TIBS combined with the MeJA system (TIBS-MeJA) were compared. Results The best immersion frequency was found to be 5/6 (5 min every 6 h) that ensured good levels of biomass and total alkaloid content in plantlets. 10 µM MeJA and 20 days of culture were the most favorable for alkaloid accumulation in the seedlings cultured in TIBS. The maximum content and productivity of total alkaloid on use of TIBS-MeJA was 2.32- and 4.69- fold, respectively, higher in terms of content, and 2.04- and 10.27- fold, respectively, higher in terms of productivity than those on use of TIBS and SSS. Conclusions Our results show that TIBS-MeJA is suitable for the large-scale production of total alkaloids in in vitro seedlings; therefore, they provide technical means for the large-scale artificial production of total alkaloids in D. nobile.

Application of Ultrasonic-Assisted Fractionation with Full-Factorial Design for the Production of Alkaloid-Rich Fraction from Ficus septica Leaves

Ficus septica Burm. F (Awar-awar) in Indonesian herbal medicine traditionally used for the treatment of various disease. Previous studies indicated that the F. septica leaves contained phenantroindolizidine alkaloids with anticancer activity. Fractionation of the ethanol extract of F. septica leaves using n-hexane was able to separate chlorophylls and other inert non–polar constituents from the extract. This fractionation process can be optimized using ultrasonic-assisted fractionation to yield n-hexane insoluble fraction (HIF) that is rich in alkaloids. This study aimed to produce HIF from the ethanol extract of F. septica leaves by using optimized ultrasonic-assisted fractionation with n-hexane as a solvent and full factorial as experimental design. Ficus septica leaves were macerated in ethanol (60%) and the extract was used for the ultrasonic-assisted fractionation process. The duration of fractionation (5-30 minutes), the extract : n-hexane ratios (1:0.1 - 1:1.66 ml/ml), and the ultrasonic power (21-106 Wrms) were optimized to determine the optimal condition for each variable. These optimal variables were used for the production of HIF from the extract by using full factorial design in the ultrasonic-assisted fractionation process. The total alkaloid content was measured using spectrophotometry and was used as parameter for the optimization process. We found that the optimal condition for the fractionation process based on each single variable optimization were 5 minutes of the fractionation duration, 66-70Wrms of the ultrasonic power, and 1:0.7 - 1:0.8 of the extract : n-hexane ratio. By using these optimized variables, the ultrasonic-assisted fractionation using full factorial design yielded up to 0.035% total alkaloid content which is almost double from those in the extract (0.019%). This study provided a basic experimental model for the production of alkaloid-enriched HIF from F. septica leaves in the industrial or pilot scale, and might contribute to the development of Indonesian herbal medicine products from natural resources.

Extraction of Alkaloids Using Ultrasound from Pulp and By-Products of Soursop Fruit (Annona muricata L.)

The main goal of this work was to measure the total alkaloid content (TALC) from pulp, peel, seed, and columella of soursop fruit (Annona muricata L.) by ultrasound-assisted extraction (UAE) and to obtain the best conditions of the UAE with the response surface methodology (RSM). We evaluated the effect of amplitude (40%, 70%, and 100%), time (5, 10, and 15 min) and pulse-cycles (0.4, 0.7, and 1 s) and compared the best UAE conditions of alkaloids with a conventional extraction (maceration). The structural characterization of the raw material with the highest TALC was developed using nuclear magnetic resonance (NMR) techniques [1H, 13C, heteronuclear single quantum correlation (HSQC), heteronuclear multiple bond correlation (HMBC), and homonuclear correlation spectroscopy (COSY)]. According to the RSM, the best conditions in the UAE for extracting alkaloids varied depending on the type of plant tissue. It took 5 min with an amplitude of 70% and pulse-cycles of 1, 0.4, and 1 s, respectively, to extract the highest TALC in peel, seed, and columella while the optimal conditions for extracting the largest amount of alkaloids from the pulp were obtained at 5 min in UAE with pulse-cycles of 0.55 s and 100% amplitude. The TALC was highest in the peel (7.48 mg/g), which was followed by the seed (2.31 mg/g), the pulp (1.20 mg/g), and the columella (0.79 mg/g) and was positively correlated (R2 = 0.98–0.88) with the predicted values. In addition, the extraction alkaloids from the peel, pulp, seed, and columella using the UAE was 56.31, 5.45, 3.06, and 2.96 times higher, respectively, than the extraction by maceration. The alkaloids identified in the peel have not been reported and were nornuciferin, assimilobin, anonaine, and isolaureline. This study showed that the soursop fruit peel can be a source of alkaloids and that UAE has an important potential for extracting these compounds.

Chromatographic Fingerprinting of the Old World Lupins Seed Alkaloids: A Supplemental Tool in Species Discrimination

The total contents and qualitative compositions of alkaloids in seeds of 10 Old World lupin species (73 accessions) were surveyed using gas chromatography. The obtained results, combined with those for three lupin crops, Lupinus angustifolius, Lupinus albus, and Lupinus luteus, provide the most complete and up-to-date overview of alkaloid profiles of 13 lupin species originating from the Mediterranean Basin. The qualitative alkaloid compositions served as useful supplementary tools of species discrimination. On the basis of the most abundant major alkaloids, lupanine, lupinine, and multiflorine, the Old World lupin species were divided into four groups. Those containing lupanine (L. angustifolius, L. albus, and Lupinus mariae-josephi), containing lupinine (Lupinus luteus, Lupinus hispanicus, and Lupinus × hispanicoluteus), containing lupinine and multiflorine (Lupinus atlanticus, Lupinus palaestinus, Lupinus anatolicus, Lupinus digitatus, Lupinus pilosus, and Lupinus cosentinii), and containing multiflorine (Lupinus micranthus). Within a given group, certain species can be, in most cases, further distinguished by the presence of other major alkaloids. The discrimination of species based on the total alkaloid content was found to be less reliable because of the significant intra-species variations, as well as the influences of environmental factors on the seed alkaloid content.

Quantitative Determination of Principal Aporphine and Benzylisoquinoline Alkaloids Due to Blooming State in Lotus Flower (Flower Buds of Nelumbo nucifera) and Their Hyaluronidase Inhibitory Activity

Using a recently developed analytical protocol, distributions of 5 aporphine alkaloids, nuciferine (1), nornuciferine (2), N-methylasimilobine (3), asimilobine (4), and pronuciferine (5), and 5 benzylisoquinoline alkaloids, armepavine (6), norarmepavine (7), N-methylcoclaurine (8), coclaurine (9), and norjuziphine (10), in lotus flowers (the flower buds of Nelumbo nucifera) were analyzed. The flowers were collected at different blooming states (beginning of blooming, one-third in bloom, half in bloom, three-quarters in bloom, and in full bloom) from Saga prefecture, Japan (NN-S1–5). The samples from the beginning of blooming state (NN-S1, 16.35 mg/g in dried material) were found to possess the richest total alkaloid content (1-10). The samples of half in bloom (NN-S3, 52.69 mg per flower of dried material) had the highest total alkaloid content per flower. Among the alkaloid constituents, nornuciferine (2, IC50 = 22.5 µM), asimilobine (4, 11.7 μM), norarmepavine (7, 26.4 μM), coclaurine (9, 11.4 μM), and norjuziphine (10, 24.3 μM) exhibited hyaluronidase inhibitory activity, which was more potent than that of the antiallergic medicine disodium cromoglycate (DSCG, 64.8 μM).