scholarly journals Extraction of Artemisinin, an Active Antimalarial Phytopharmaceutical from Dried Leaves of Artemisia annua L., Using Microwaves and a Validated HPTLC-Visible Method for Its Quantitative Determination

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Himanshu Misra ◽  
Darshana Mehta ◽  
Bhupendra Kumar Mehta ◽  
Dharam Chand Jain
Keyword(s):  
Thin Layer ◽  
Artemisia Annua ◽  
Main Stem ◽  
Thin Layer Plate ◽  
Growth Stages ◽  
Root Extract ◽  
Hptlc Plate ◽  
Pharmaceutical Industries ◽  
Artemisinin Content

A simple, rapid, precise, and accurate high-performance thin-layer chromatographic method coupled with visible densitometric detection of artemisinin is developed and validated. Samples of the dried Artemisia annua leaves were extracted via microwaves using different solvents. This method shows the advantage of shorter extraction time of artemisinin from leaves under the influence of electromagnetic radiations. Results obtained from microwave-assisted extraction (MAE) were compared with hot soxhlet extraction. Chromatographic separation of artemisinin from plant extract was performed over silica gel 60 F254 HPTLC plate using n-hexane : ethyl acetate as mobile phase in the ratio of 75 : 25, v/v. The plate was developed at room temperature 25 ± 2.0°C. Artemisinin separation over thin-layer plate was visualized after postchromatographic derivatization with anisaldehyde-sulphuric acid reagent. HPTLC plate was scanned in a CAMAG’s TLC scanner 3 at 540 nm. Artemisinin responses were found to be linear over a range of 400–2800 ng spot−1 with a correlation coefficient 0.99754. Limits of detection and quantification were 40 and 80 ng spot−1, respectively. The HPTLC method was validated in terms of system suitability, precision, accuracy, sensitivity (LOD and LOQ), and robustness. Additionally, calculation of plate efficiency and flow constant were included as components of validation. Extracts prepared from different parts of the plant (leaves, branches, main stem, and roots) were analyzed for artemisinin content, in which, artemisinin content was found higher in the leaf extract with respect to branches and main stem extracts; however, no artemisinin was detected in root extract. The developed HPTLC-visible method of artemisinin determination will be very useful for pharmaceutical industries, which are involved in monitoring of artemisinin content during different growth stages (in vitro and in vivo) of A. annua for qualitative and quantitative assessment of final produce prior to commercial-scale processing for assessment of cost-benefit ratio.

scholarly journals Antitrypanosomal and Antileishmanial Activities of Organic and Aqueous Extracts of Artemisia Annua

2008 ◽  
Vol 3 (12) ◽  
pp. 1934578X0800301
Author(s):  
Anna Rita Bilia ◽  
Marcel Kaiser ◽  
Franco Francesco Vincieri ◽  
Deniz Tasdemir
Keyword(s):  
Artemisia Annua ◽  
Antimalarial Activity ◽  
Antiparasitic Activity ◽  
Growth Inhibitory ◽  
Growth Inhibitory Activity ◽  
Organic Extracts ◽  
Antileishmanial Activities ◽  
Artemisinin Content ◽  
Protozoan Species

Artemisia annua is an herbal drug with profound antimalarial activity, which can be ascribed to the sesquiterpene lactone artemisinin. Artemisinin also shows efficacy against other parasitic protozoan species, such as Trypanosoma and Leishmania, however trypanocidal and leishmanicidal effects of A. annua extracts have not been reported so far. In the current study, we evaluated the in vitro growth inhibitory activity of a number of organic and aqueous A. annua extracts, including tinctures, infusions and decoctions against three parasitic protozoa, T. brucei rhodesiense, T. cruzi and L. donovani. Artemisinin content of these extracts was determined by HPLC/DAD/MS. Artemisinin was also evaluated for its antiparasitic activity for comparison. Among the tested extracts, the acetone- and the n-hexane-solubles of A. annua were the most potent against T. b. rhodesiense with IC50 values of 0.30 and 0.455 μg/mL, respectively, whereas the other extracts were ten- to fifty-fold less potent. Neither of the extracts nor artemisinin had trypanocidal activity against T. cruzi (IC50 > 30 μg/mL). Only the organic extracts of A. annua arrested the growth of L. donovani with modest IC50 values (5.1 to 9.0 μg/mL) comparable to that of artemisinin (IC50 8.8 μg/mL). This study highlights significant variations in the artemisinin content of A. annua extracts and underlines the potential of A. annua extracts and artemisinin in the treatment of trypanosomal and leishmanial infections.

scholarly journals In vitro reduction of Plasmodium falciparum gametocytes by Artemisia spp. tea infusions

2020 ◽  
Author(s):  
Danielle Snider ◽  
Pamela J. Weathers
Keyword(s):  
Gene Expression ◽  
Clinical Trial ◽  
Plasmodium Falciparum ◽  
Late Stage ◽  
Artemisia Annua ◽  
Clinical Results ◽  
Marker Genes ◽  
Tea Infusions ◽  
Artemisinin Content

AbstractIn this study, we showed in vitro evidence that supports the efficacy of A. annua and A. afra tea infusions used in a 2015 clinical trial not only for clearing asexual Plasmodium falciparum parasites, but also for eliminating sexual gametocytes. P. falciparum NF54 was grown in vitro, synchronized, and induced to form gametocytes using N-acetylglucosamine. Cultures during asexual, early, and late stage gametogenesis were treated with artemisinin, methylene blue, and Artemisia annua and A. afra tea infusions (5g DW/L) using cultivars that contained 0-283 μM artemisinin. Asexual parasitemia and gametocytemia were analyzed microscopically. Gametocyte morphology was also scored. Markers of early (PfGEXP5) and late stage (Pfs25) gametocyte gene expression were also measured using RT-qPCR. Both A. annua and A. afra tea infusions reduced gametocytemia in vitro, and the effect was mainly artemisinin dependent. Expression levels of both marker genes were reduced with the effect mainly attributed to artemisinin content of the our tested Artemisia cultivars. Tea infusions of both species also inhibited asexual parasitemia and although mainly artemisinin dependent, there was a weak antiparasitic effect from artemisinin-deficient A. afra. These results showed that A. annua and to a lesser extent, A. afra, inhibited parasitemia and gametogenesis in vitro, and results are consistent with earlier observed clinical results.

scholarly journals Differentially Expressed Genes during Contrasting Growth Stages of Artemisia annua for Artemisinin Content

PLoS ONE ◽  
2013 ◽  
Vol 8 (4) ◽  
pp. e60375 ◽  
Author(s):  
Priya Nair ◽  
Amita Misra ◽  
Alka Singh ◽  
Ashutosh K. Shukla ◽  
Madan M. Gupta ◽  
...  
Keyword(s):  
Differentially Expressed Genes ◽  
Artemisia Annua ◽  
Differentially Expressed ◽  
Growth Stages ◽  
Artemisinin Content

scholarly journals In vitro alteration of artemisinin biosynthesis in Artemisia annua L during treatment with methyl jasmonate

2020 ◽  
Vol 19 (1) ◽  
pp. 33-37
Author(s):  
Pravej Alam ◽  
Thamer H. Albalawi
Keyword(s):  
Methyl Jasmonate ◽  
Plant Height ◽  
Artemisia Annua ◽  
Dry Weight ◽  
Artemisia Annua L ◽  
Biomass Plant ◽  
Vitro Effect ◽  
Artemisinin Biosynthesis ◽  
Artemisinin Content

Purpose: To investigate the in vitro effect of methyl jasmonate (MeJA) on artemisinin biosynthesis in Artemisia annua.Method: The effect of two concentrations of MeJA i.e. 2 μm (T2) and 5 μm (T5) on biomass, plant height and artemisinin content of Artemisia annua L. was investigated in vitro in MS medium.Results: Plant height (6.5 ± 1.8 cm), biomass (0.18 ± 0.02 g/plant) and artemisinin content (0.035 ± 0.002 % dry weight) were higher in T5 and T2 treatment groups than in non-treated control plants (C). Artemisinin content, biomass and plant height were positively correlated after 2 - 8 days of treatment, but remained unchanged on the 12th day of treatment.Conclusion: The observed effects of exogenous MeJA on the biosynthesis of artemisinin and other secondary metabolites may lead to elucidation of promising targets for further studies on metabolic engineering of Artemisia annua L. Keywords: Artemisia annua L. Methyl jasmonate, HPLC, Artemisinin

scholarly journals Associating gene expressions with curcuminoid biosynthesis in turmeric

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Dipendra Kumar Ayer ◽  
Kaushal Modha ◽  
Vipulkumar Parekh ◽  
Ritesh Patel ◽  
Gopal Vadodariya ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Patterns ◽  
Negative Association ◽  
Growth Stages ◽  
Gene Expressions ◽  
Individual Gene ◽  
Pharmaceutical Industries ◽  
The Impact

Abstract Background Biologically important curcuminoids viz curcumin, demethoxycurcumin, and bisdemethoxycurcumin in turmeric rhizome have several health benefits. Pharmaceutical industries synthesize curcuminoids manipulating gene expressions in vitro or in vivo because of their medicinal importance. In this experiment, we studied the gene expressions involved in the curcuminoid biosynthesis pathway in association with curcuminoid yield in turmeric rhizome to study the impact of individual gene expression on curcuminoid biosynthesis. Results Gene expressions at the different growth stages of turmeric rhizome were association tested with respective curcuminoid contents. Gene expression patterns of diketide-CoA synthase (DCS) and multiple curcumin synthases (CURSs) viz curcumin synthase 1 (CURS1), curcumin synthase 2 (CURS2), and curcumin synthase 3 (CURS3) were differentially associated with different curcuminoid contents. Genotype and growth stages showed a significant effect on the gene expressions resulting in a significant impact on curcuminoid balance in turmeric rhizome. DCS and CURS3 expression patterns were similar but distinct from CURS1 and CURS2 expression patterns in turmeric rhizome. DCS expression had a significant positive and CURS3 expression had a significant negative association with curcumin, demethoxycurcumin, bisdemethoxycurcumin, and total curcuminoid in turmeric rhizome. CURS1 expression had a negative association with curcumin whereas CURS2 expression showed a positive association with demethoxycurcumin. Conclusions The effects of DCS and CURS expressions are not always positive with different curcuminoid contents in turmeric rhizome. DCS expression has a positive and CURS3 expression has a negative association with curcuminoids. CURS1 and CURS2 are also associated with curcumin and demethoxycurcumin synthesis. This mechanism of co-expression of diketide-CoA synthase and multiple curcumin synthases in turmeric rhizome has a significant effect on curcuminoid balance in different turmeric cultivars. Further experiment will explore more insights; however, association-based test results from this experiment can be useful in improving curcuminoid yield in turmeric rhizome or in vitro through the application of genetic engineering and biotechnology. Graphical abstract Associating gene expressions with curcuminoid biosynthesis in turmeric

Antimutagenic, Antiproliferative and Antioxidant Properties of Sea Grape Leaf Extract Fractions (Coccoloba uvifera L.)

2021 ◽  
Vol 21 ◽  
Author(s):  
Jorge A. Ramos-Hernández ◽  
Montserrat Calderón-Santoyo ◽  
Armando Burgos-Hernández ◽  
Joel S. García- Romo ◽  
Arturo Navarro-Ocaña ◽  
...  
Keyword(s):  
Antioxidant Properties ◽  
In Vitro Tests ◽  
Antimutagenic Activity ◽  
Hexane Extraction ◽  
Biological Potential ◽  
Hct 116 ◽  
Pharmaceutical Industries ◽  
Grape Leaf ◽  
First Time

Background: Cancer is a disease characterized by the invasion and uncontrolled growth of cells. One of the best ways to minimize the harmful effects of mutagens is through the use of natural antimutagens. In this regard, the search for new antimutagens that act in the chemoprevention could represent a promising field in this area. Objective: In this study biological potential of 11 fractions from Coccoloba uvifera L. leaf hexane extract was evaluated by several in vitro tests. Methods: Leaves were lyophilized and hexane extraction was performed. The extract was fractionated by column chromatography with hexane, ethyl acetate, and methanol. The antimutagenic (Ames test), antiproliferative (MTT test), and antioxidant capacity (DPPH, ABTS, and ferrous ion chelation) of the fractions were evaluated. Results: Fractions 4, 6, 8, and 9 have antimutagenic activity (against sodium azide in strain TA100), fraction 11 showed antiproliferative capacity (IC50 of 24 ± 9 μg/mL in cells of HCT 116). The fractions with the highest activity were analyzed by HPLC-MS and lupeol, acacetin, and β-sitosterol were identified. Conclusion: This study demonstrates, for the first time, the bioactivity of C. uvifera leaf as a new source of high biological value compounds (HBVC), which can be of interest to the food and pharmaceutical industries.

Phytochemical, Analytical and Medicinal Studies of Holoptelea integrifolia Roxb. Planch - A Review

2019 ◽  
Vol 5 (4) ◽  
pp. 270-277 ◽  
Author(s):  
Vijay Kumar ◽  
Simranjeet Singh ◽  
Ragini Bhadouria ◽  
Ravindra Singh ◽  
Om Prakash
Keyword(s):  
Liquid Chromatography ◽  
Thin Layer ◽  
Thin Layer Chromatography ◽  
High Performance ◽  
Biologically Active ◽  
Toxicological Studies ◽  
Wide Range ◽  
Layer Chromatography

Holoptelea integrifolia Roxb. Planch (HI) has been used to treat various ailments including obesity, osteoarthritis, arthritis, inflammation, anemia, diabetes etc. To review the major phytochemicals and medicinal properties of HI, exhaustive bibliographic research was designed by means of various scientific search engines and databases. Only 12 phytochemicals have been reported including biologically active compounds like betulin, betulinic acid, epifriedlin, octacosanol, Friedlin, Holoptelin-A and Holoptelin-B. Analytical methods including the Thin Layer Chromatography (TLC), High-Performance Thin Layer Chromatography (HPTLC), High-Performance Liquid Chromatography (HPLC) and Liquid Chromatography With Mass Spectral (LC-MS) analysis have been used to analyze the HI. From medicinal potency point of view, these phytochemicals have a wide range of pharmacological activities such as antioxidant, antibacterial, anti-inflammatory, and anti-tumor. In the current review, it has been noticed that the mechanism of action of HI with biomolecules has not been fully explored. Pharmacology and toxicological studies are very few. This seems a huge literature gap to be fulfilled through the detailed in-vivo and in-vitro studies.

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