scholarly journals PHYTOCHEMICAL ANALYSIS OF CALLUS TWO VARIETIES ORTHOSIPHON ARISTATUS (BLUME) MIQ ON MURASHIGE AND SKOOG MEDIA: A STRATEGIC STEP OF SECONDARY METABOLITE PRODUCTION

2021 ◽  
pp. 71-77
Author(s):  
FAHRAUK FARAMAYUDA ◽  
TOTIK SRI MARIANI ◽  
ELFAHMI ◽  
SUKRASNO
Keyword(s):  
Secondary Metabolites ◽  
Rosmarinic Acid ◽  
High Performance ◽  
Leaf Explants ◽  
Gradient Elution ◽  
Growth Media ◽  
Phytochemical Analysis ◽  
New Information ◽  
Sterile Leaf

Objective: The research aimed to provide new information regarding the secondary metabolites content of purple and white-purple Orthosiphon aristatus (Blume) Miq. callus, which can then be used as a basis for developing towards cell suspension and ultimately producing secondary metabolites using bioreactors. Methods: Callus induction of two varieties of O. aristatus were performed by inoculating sterile leaf explants grown on Murashige and Skoog basal media supplemented with 2,4-dichlorophenoxyacetis acid 0.4 ppm. The secondary metabolites were analysed and quantified using high-performance liquid chromatography with gradient elution. Results: The results showed the growth of callus two varieties of O. aristatus in growth media MS with 2,4-D 0.4 ppm. Rosmarinic acid content in the acetone extract of the purple variety callus was 1.28% w/w, and the white-purple variety was 2.22% w/w. Conclusion: This study could form the basis for the development of rosmarinic acid production by In vitro culture modification.

scholarly journals Cytokinin Mediated Increased In Vitro Production of Secondary Metabolites with Special Reference to Solasodine in Solanum erianthum

2021 ◽  
Vol 8 (02) ◽  
pp. e62-e68
Author(s):  
Jeeta Sarkar ◽  
Nirmalya Banerjee
Keyword(s):  
Secondary Metabolites ◽  
High Performance ◽  
Culture Medium ◽  
In Vitro Production ◽  
Leaf Extracts ◽  
Plant Parts ◽  
Regenerated Plants ◽  
Vitro Production ◽  
Regenerated Plantlets

AbstractSteroid alkaloid solasodine is a nitrogen analogue of diosgenin and has great importance in the production of steroidal medicines. Solanum erianthum D. Don (Solanaceae) is a good source of solasodine. The aim of this study was to evaluate the effect of different cytokinins on the production of secondary metabolites, especially solasodine in the in vitro culture of S. erianthum. For solasodine estimation, field-grown plant parts and in vitro tissues were extracted thrice and subjected to high-performance liquid Chromatography. Quantitative analysis of different secondary metabolites showed that the amount was higher in the in vitro regenerated plantlets compared to callus and field-grown plants. The present study critically evaluates the effect of the type of cytokinin used in the culture medium on solasodine accumulation in regenerated plants. The highest solasodine content (46.78±3.23 mg g-1) was recorded in leaf extracts of the in vitro grown plantlets in the presence of 6-γ,γ-dimethylallylamino purine in the culture medium and the content was 3.8-fold higher compared to the mother plant.

scholarly journals In vitro molluscicidal activity of wormwood (Artemisia dubia) leaves against Oncomelania hupensis quadrasi

2019 ◽  
pp. 16-23 ◽  
Author(s):  
Charles Jardian Alinsub ◽  
Melvin Bagot
Keyword(s):  
Secondary Metabolites ◽  
Methanolic Extract ◽  
Positive Control ◽  
Phytochemical Analysis ◽  
Snail Population ◽  
Artemisia Dubia ◽  
Negative Controls ◽  
Infusion Technique ◽  
Molluscicidal Activity

There have been many efforts to eradicate the problem of schistosomiasis. One method is to control the growing snail population. This study aimed to determine the molluscicidal activity of wormwood (Artemisia dubia)leaves methanolic extract against adult and juvenile. Using infusion technique, 80% methanol was used as solvent. There were eight treatments used: distilled water (T0(-)1) and 1% methanol (T0(-)2) as negative controls; 0.0002% (2mg/L) niclosamide (T0(+)) as positive control; and 11.121% (T1), 12.478% (T2), 14% (T3), 15.708% (T4) and 17.625% (T5), which were replicated five times with 10 snails per replicate. Results showed that for adults, the wormwood leaves methanolic extract was not statistically significant with the commercially available molluscicide, although 17.625% (T5) resulted in 82% mortality rate. For juveniles, the different extract concentrations resulted in 98 to 100% mortality, which were comparable to 0.0002% (2mg/L) niclosamide and were considered highly effective based on Reik & Keitz (1954) and European Agency for the Evaluation of Medicinal Products (EMEA). Thus, wormwood leaves methanolic extract has a promising molluscicidal activity against adult snails at a concentration of 17.625%, and at all concentrations for the juvenile snails of O. hupensis quadrasi. The LC50 of the adult and juvenile snails were 14.075% and 10.294%, respectively. Secondary metabolites that were found positive during the qualitative phytochemical analysis with the extract were tannins, saponins, and terpenoids. The mortality of the snails can be attributed to the bioactivity of the secondary metabolites present that may be acting in combination or individually.

scholarly journals Effect Effect of 2,4-D and BA on the establishment of cell suspension from nodes and internodes of Capsicum annuum cv. Etna

2019 ◽  
Vol 7 (5) ◽  
pp. 55-61
Author(s):  
MAURICIO REGINALDO ALVES DOS SANTOS ◽  
CAROLINA SOUZA
Keyword(s):  
Secondary Metabolites ◽  
Cell Suspension ◽  
Capsicum Annuum ◽  
Growth Pattern ◽  
Cellular Proliferation ◽  
Biological Activities ◽  
Leaf Explants ◽  
Deceleration Phase ◽  
A Cell

In vitro cell suspension cultivation systems have been largely reported as safe and standardized methods for production of secondary metabolites with medicinal and agricultural interest. Capsicum annuum is one of the most widely grown vegetable in the world and its biological activities have been demonstrated against insects, fungi, bacteria and other groups of organisms. The determination of procedures for the dedifferentiation of cells into callus cells and the subsequent study of the callus growth pattern are necessary for the establishment of cell suspensions and also to subsidize studies regarding the bioactivity of its secondary metabolites. The objective of this study was to establish a protocol for dedifferentiation of leaf cells of the cultivar C. annuum cv. Etna and to determine the growth pattern of the calluses with a focus on the deceleration phase, when the callus cells must be subcultured into a liquid medium in order to establish cell suspension cultivations aiming at the production of secondary metabolites. treatment that resulted in the highest %CI, ACCC and callus weight was the combination of 4.52 µM 2,4-D + 0.44 µM BA. The calluses produced were friable and whitish and their growth pattern followed a sigmoid shape. The deceleration phase started on the 23rd day of cultivation. Callus induction in leaf explants of C. annuum cv. Etna can be achieved in MS medium supplemented with 4.52 µM 2,4-D + 0.44 µM BA, which results in high cellular proliferation; in order to start a cell suspension culture, callus cells on the 23rd day of culture should be used.

HPTLC analysis and in vitro antioxidant activity of aqueous bark extract of Erythrina variegata L.

2016 ◽  
Vol 63 (3) ◽  
pp. 143-157 ◽  
Author(s):  
Kanakasabapathy Devaki
Keyword(s):  
Antioxidant Activity ◽  
Secondary Metabolites ◽  
High Performance ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Bark Extract ◽  
Erythrina Variegata ◽  
High Antioxidant Activity ◽  
Layer Chromatography

Erythrina variegata L. is an important medicinal plant used in the preparations of Ayurvedic formulations used against several ailments. This study was carried out to investigate the presence of secondary metabolites using phytochemical screening, high-performance thin-layer chromatography (HPTLC) fingerprinting analysis and the antioxidant potential of the aqueous bark extract of E. variegata L. The secondary metabolites and the free radical scavenging activity were analyzed using standard protocols. The results obtained in the present study revealed that E. variegata has high antioxidant activity against free radicals based on phytoconstituents.

scholarly journals Micropropagation of Eryngium campestre L. via Shoot Culture Provides Valuable Uniform Plant Material with Enhanced Content of Phenolic Acids and Antimicrobial Activity

2016 ◽  
Vol 58 (1) ◽  
pp. 43-56 ◽  
Author(s):  
Małgorzata Kikowska ◽  
Barbara Thiem ◽  
Elwira Sliwinska ◽  
Monika Rewers ◽  
Mariusz Kowalczyk ◽  
...  
Keyword(s):  
Phenolic Acids ◽  
Rosmarinic Acid ◽  
High Performance ◽  
Nuclear Dna ◽  
Plant Biomass ◽  
Shoot Culture ◽  
Trichophyton Mentagrophytes ◽  
Inhibitory Effect ◽  
Tlc Analysis

AbstractAn efficient micropropagation protocol for production of genetically uniform clones ofEryngium campestreL. was developed. To determine the effect of nutritional and hormonal factors on shoot and root development and bioactive compounds production, three variants of media differing in the content of macro- and micronutrients, as well as plant growth regulators of various types and concentrations were tested. The highest regeneration (100%), with over 13 shoots per explant, was induced on Murashige and Skoog (MS) medium with 1.0 mg l−1benzyladenine (BA) and 0.1 mg l−1indole-3-acetic acid (IAA). The in vitro derived shoots multiplied through axillary bud formation were rooted and transferred to an experimental plot with 78% frequency of survival. Flow cytometry showed no variation in nuclear DNA between the seedlings and micropropagated plants. Preliminary thin layer chromatography (TLC) analysis indicated that phenolic acids, saponins, flavonoids and acetylenes were present in plant biomass. Ultra high performance liquid chromatography (UHPLC) analysis revealed that shoots and roots from in vitro derived plants and root cultures maintained the ability to produce rosmarinic acid (RA), rosmarinic acid hexoside (RA-HEX) and chlorogenic acid (CGA). The highest phenolic acid content was detected in roots of in vitro regenerated plants. The extract from those roots expressed the highest inhibitory effect against bacteriaStaphylococcus aureus, as well as dermatophytesTrichophyton mentagrophytesandT. rubrum.

scholarly journals RAPID IN VITRO CALLOGENESIS AND PHYTOCHEMICAL SCREENING OF LEAF, STEM AND LEAF CALLUS OF MUSSAENDA FRONDOSA LINN. - A MEDICINAL PLANT.

2017 ◽  
Vol 10 (6) ◽  
pp. 81 ◽  
Author(s):  
Manasa Dj ◽  
Chandrashekar Kr ◽  
Bhagya N
Keyword(s):  
Methanolic Extract ◽  
Antioxidant Activities ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Leaf Explants ◽  
Reducing Power ◽  
Callus Cultures ◽  
Phytochemical Analysis ◽  
Qualitative And Quantitative

Objective: To standardise the protocol for rapid callogenesis in Mussaenda frondosa L. using leaf explants. Qualitative and quantitative phytochemical analysis of leaf, stem and callus cultures.Methods: The leaf explants were inoculated onto MS medium supplemented with varying concentrations of growth regulators such as 2, 4 - D, NAA, BAP, Kn for the induction of callus. Qualitative and quantitative analysis of total phenol, flavonoids and alkaloids contents of leaf, stem and callus were tested by standard methods.  The antioxidant activities were investigated using DPPH radical scavenging method and reducing power assay. The anti - inflammatory activity was evaluated by membrane stabilizing activity.Results: Pale green, healthy, friable and fast growing callus was obtained on the medium enriched with NAA (2mg/l) + Kn (4mg/l). Quantitative determination showed the highest concentration of total phenolics in the methanolic extract of in vitro grown callus (10 ± 1.1 mg of GA/g of extract), flavonoids in methanolic stem extract (137±1.6 mg of Quercitin/g of extract) and alkaloids in methanolic extract of leaf (118.3±1.5 mg/10g of extract). The methanolic leaf extract exhibited highest free radical scavenging activity with IC50 value of 40.6±10.06 μg/ml. The highest membrane stabilizing activity was shown by chloroform extract of the leaf (66.02%).Conclusion: The present preliminary phytochemical and pharmacological analysis may form the basis for drug development in future using callus cultures of M. frondosa.   

scholarly journals Induction, biochemical trait and phytochemical screening of calluses of Amburana cearensis (Allemão) A.C. Smith

2020 ◽  
Vol 42 ◽  
pp. e54187
Author(s):  
Jéssica Nascimento Costa Vascocelos ◽  
Alone Lima Brito ◽  
Andressa Priscila Pianco Santos Lima ◽  
Jackson Roberto Guedes da Silva Almeida ◽  
Ana Paula de Oliveira ◽  
...  
Keyword(s):  
Growth Curve ◽  
High Performance ◽  
Reducing Sugars ◽  
Dichlorophenoxyacetic Acid ◽  
Phytochemical Analysis ◽  
Phytochemical Screening ◽  
Total Proteins ◽  
Biochemical Tests ◽  
2,4 Dichlorophenoxyacetic Acid

Amburana cearensis is an arboreal legume of the Fabaceae family, with high phytotherapic and medicinal potential due the presence of secondary metabolites. The objective of this study was to evaluate the effect of 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-amino-2,5,6- trichloro-2-pyridinecarboxylic acid (picloram) on the in vitro induction of callogenesis of A. cearensis and analyze the biochemical and phytochemical potential of these calluses. For callus induction, leaf and cotyledon segments were used as explants, which were inoculated in woody plant medium (WPM) supplemented with different concentrations of 2,4-D (0, 5, 10, 20, 40 μM) or picloram (0, 5, 10, 20, 40, 80 μM). The callus growth curve was estimated based on fresh weight, measured at 7-day intervals until 28 days after inoculation. The calluses were analyzed by biochemical tests to quantify the reducing sugars and total proteins. Phytochemical screening and high-performance liquid chromatography were performed to establish the phytochemical profile of extracts from calluses. The concentrations of 21.94 μM and 26.46 μM of 2,4-D induced the greatest formation of compact and friable calluses from the leaf and cotyledon segments, respectively. The growth curve had two distinct phases (lag and exponential) for both types of calluses evaluated. The maximum levels of reducing sugars and total proteins in the calluses from leaf and cotyledon segments were obtained on the day of inoculation and after 28 days of cultivation, respectively. The results of the phytochemical analysis identified the presence of coumarin in all the extracts evaluated, this secondary metabolite has high pharmacological potential.

In vitro organogenesis secondary metabolite production and heavy metal analysis in Swertia chirayita

2014 ◽  
Vol 9 (7) ◽  
pp. 686-698 ◽  
Author(s):  
Vijay Kumar ◽  
Shailesh Singh ◽  
Rajib Bandopadhyay ◽  
Madan Sharma ◽  
Sheela Chandra
Keyword(s):  
Heavy Metal ◽  
Secondary Metabolites ◽  
Secondary Metabolite ◽  
Leaf Explants ◽  
Secondary Metabolite Production ◽  
Direct Organogenesis ◽  
Metabolite Production ◽  
Swertia Chirayita

AbstractAn efficient protocol of plant regeneration through direct and indirect organogenesis in Swertia chirayita was developed. Explants cultured on Murashige and Skoog medium supplemented with 2,4-D (0.5 mg L−1) with combination of Kinetin (0.5 mg L−1) showed the highest frequency (84%) of callusing and 1.0mg L−1 6-benzyladenine (BA) in combination with (100 mg L−1) Adenine sulphate (Ads) + (0.1 mg L−1) Indole acetic acid (IAA) was excellent for maximum adventitious shoot (12.69 ± 1.30) formation in four week of culture. A maximum number of (7.14 ± 0.99) shoots were developed per leaf explants through direct organogenesis. The highest frequency of rooting (11.46 ± 1.56) was observed on MS medium augmented with IAA (1.0 mg L−1). Well-rooted shoots transferred to plastic pots containing a soilrite: sand mix and then moved to the greenhouse for further growth and development. Four major secondary metabolites were analyzed and quantified using high performance liquid chromatography. Amount of secondary metabolites was found significantly higher, in in vitro plantlets compared to in vivo plantlets and callus raised from S. chirayita. Higher heavy metal accumulation in in vitro as compared to in vivo plantlets correlates higher secondary metabolite production supporting that they play regulatory role in influencing the plant secondary metabolism.

scholarly journals Determination of Tedizolid in Bacterial Growth Medium Mueller-Hinton Broth by High-Performance Liquid Chromatography and Its Application to an In Vitro Study in the Hollow-Fiber Infection Model

Separations ◽  
2021 ◽  
Vol 8 (9) ◽  
pp. 141
Author(s):  
Khalid Iqbal ◽  
Aliki Milioudi ◽  
Elena Haro Martínez ◽  
Sebastian Georg Wicha
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Growth Medium ◽  
Bacterial Growth ◽  
High Performance ◽  
Infection Model ◽  
Growth Media ◽  
Mueller Hinton ◽  
Mueller Hinton Broth

Pharmacokinetic/pharmacodynamic (PKPD) studies of anti-infectives are frequently performed in in vitro infection models where accurate quantification of antibiotic concentrations in bacterial growth media is crucial to establish PK/PD relationships. Here, a sensitive and rapid high-performance liquid chromatography (HPLC) method was developed to quantify tedizolid (TDZ) in the bacterial growth medium Mueller-Hinton broth (MHB). Matrix components were separated by direct protein precipitation with methanol (1:1). The chromatographic separation was carried out in a Dionex Ultimate 3000 HPLC system using an Accucore® C-18 RPMS HPLC column (2.6 µm, 100 × 2.1 mm) using isocratic elution with 25% acetonitrile and 75% of 0.1% formic acid. The lower limit of quantification was 0.03 mg/L when measured at 300 nm. Following relevant European Medicine Agency guidelines, the method was successfully validated for linearity, selectivity, recovery, inter- and intra-day precision, and accuracy and stability. When applied to in vitro PKPD studies, the method successfully quantified a range of TDZ concentration (Cmin, 0.09-Cmax, 0.65 mg/L) in MHB. The analyzed concentrations were in line with the planned PK profiles. The application of the developed method to quantify TDZ in MHB in in vitro PKPD studies is warranted.

scholarly journals EVALUATION OF HEPATOPROTECTIVE POTENTIAL OF LEAF AND LEAF CALLUS EXTRACTS OF ANISOCHILUS CARNOSUS (L) WALL.

2018 ◽  
Vol 10 (3) ◽  
pp. 156
Author(s):  
Nissar Ahmad Reshi ◽  
Sudarahana Mysore Shankarsingh ◽  
Girish Hodiyala Vasanaika
Keyword(s):  
Secondary Metabolites ◽  
Cell Viability ◽  
Hepg2 Cell ◽  
Hepg2 Cells ◽  
Alcohol Intoxication ◽  
Leaf Explants ◽  
Hepatoprotective Activity ◽  
Phytochemical Analysis ◽  
Toxic Dose ◽  
Viability Percentage

<p>The study was carried out to evaluate the hepatoprotective activity of leaf and leaf callus extracts of <em>Anisochilus carnosus</em> (L) Wall. against alcohol induced toxicity using HepG2 cell line. Leaf explants were cultured on Murashige and Skoog solid medium supplemented with different growth regulators. Prior to the determination of hepatoprotective property leaf and leaf callus extracts were subjected to the toxic dose study. The degree of hepatoprotection of extracts was determined by measuring cell viability percentage by MTT assay. The preliminary phytochemical analysis of leaf and leaf callus was carried out by qualitative analysis. Maximum percentage of callus formation (98%) was obtained in MS medium fortified with 3 mg/l 2,4-D. HepG2 cells were pretreated with the different concentrations (below toxic dose) of leaf and leaf callus extracts for 72 hours followed by alcohol intoxication. Results revealed that ethanolic leaf extract pretreated HepG2 cells show 94% cell viability compared to the standard silymarin pretreated HepG2 cells which showed 81% cell viability. Leaf callus extracts also exhibited significant hepatoprotective activity where ethanolic callus extract pretreated HepG2 cells showed 86% viability after intoxication with alcohol. Results revealed that HepG2 cell viability percentage is dose dependent. Phytochemical studies revealed the presence of different secondary metabolites in leaf and leaf callus extracts. The bio-efficacy study confirms the presence of secondary metabolites of hepatoprotective nature in leaf and leaf callus of <em>A. carnosus.</em></p>

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