1. The project aims to isolate and characterize berberine from Anamirta cocculus stem through ultrasonication assisted extraction. The biological activities of the extracts will also be evaluated.
2. Berberine will be isolated through column chromatography and characterized using spectroscopy. Antioxidant, anti-inflammatory, and antimicrobial activities of the extracts and isolated berberine will be determined.
3. The project involves preparation of stem extracts, isolation of berberine, and evaluation of antioxidant, anti-inflammatory, and antimicrobial properties to understand the biological potential of Anamirta cocculus stem.
what is extraction. what are the conventional methods what are their drawbacks. what are the advanced techniques used to overcome those drawbacks. major focus on Microwave assisted extraction. its factors advantages types and applications
what is extraction. what are the conventional methods what are their drawbacks. what are the advanced techniques used to overcome those drawbacks. major focus on Microwave assisted extraction. its factors advantages types and applications
INDUSTRIAL PHARMACOGNOSTICAL TECHNOLOGY
Herbal drug industry: Infrastructure of herbal drug industry
involved in production of standardized extracts and various
dosage forms. Current challenges in upgrading and
modernization of herbal formulations. Entrepreneurship
Development, Project selection, project report, technical
knowledge, Capital venture, plant design, layout and construction.
Pilot plant scale –up techniques, case studies of herbal extracts.
Formulation and production management of herbals.
Extraction is a process of separation or isolation of pharmaceutical active ingredients
from plant or animal drugs with the help of solvent.
On the basis of the physical nature of crude drug to be extracted i.e. liquid or solid ,the extraction process may be:
Liquid –Liquid Extraction Or
Solid –Liquid Extraction.
The solvent used for extraction is called as ‘Menstruum’ and the residue left after extracting desired constituents is called ‘Marc’.
Required Ideal Properties of Menstruum :
Should be inert and non –toxic
Should extract only the desirable constituent of the crude drug .
Should be cheap and easily available
INDIAN AND INTERNATIONAL PATENT LAW AS APPLICABLE TO HERBAL AND NATURAL PRODUCTSTejaswini Chandra
ITS A USEFUL TOPIS FOR LAW STUDNETS REGARDING THE HERBAL DRUGS AND NATURAL PRODUCTS .
ALSO USEFUL FOR THE MPHARM STUDENTS OF ALL DOMAIN TO STUDY REGARDING THE PATENT LAW
HOPE YOU ALL MAKE USE OF IT TO FULLEST AND SUCCEED.
method to separate compounds based on their relative solubilities in two different immiscible liquids, usually water and an organic solvent. It is an extraction of a substance from one liquid into another liquid phase.
coumarin; umbelliferone and its biosynthesis and isolation.
terpenoide; cucurbitacine and its biosynthesis and isolation purification and characterization
INDUSTRIAL PHARMACOGNOSTICAL TECHNOLOGY
Herbal drug industry: Infrastructure of herbal drug industry
involved in production of standardized extracts and various
dosage forms. Current challenges in upgrading and
modernization of herbal formulations. Entrepreneurship
Development, Project selection, project report, technical
knowledge, Capital venture, plant design, layout and construction.
Pilot plant scale –up techniques, case studies of herbal extracts.
Formulation and production management of herbals.
Extraction is a process of separation or isolation of pharmaceutical active ingredients
from plant or animal drugs with the help of solvent.
On the basis of the physical nature of crude drug to be extracted i.e. liquid or solid ,the extraction process may be:
Liquid –Liquid Extraction Or
Solid –Liquid Extraction.
The solvent used for extraction is called as ‘Menstruum’ and the residue left after extracting desired constituents is called ‘Marc’.
Required Ideal Properties of Menstruum :
Should be inert and non –toxic
Should extract only the desirable constituent of the crude drug .
Should be cheap and easily available
INDIAN AND INTERNATIONAL PATENT LAW AS APPLICABLE TO HERBAL AND NATURAL PRODUCTSTejaswini Chandra
ITS A USEFUL TOPIS FOR LAW STUDNETS REGARDING THE HERBAL DRUGS AND NATURAL PRODUCTS .
ALSO USEFUL FOR THE MPHARM STUDENTS OF ALL DOMAIN TO STUDY REGARDING THE PATENT LAW
HOPE YOU ALL MAKE USE OF IT TO FULLEST AND SUCCEED.
method to separate compounds based on their relative solubilities in two different immiscible liquids, usually water and an organic solvent. It is an extraction of a substance from one liquid into another liquid phase.
coumarin; umbelliferone and its biosynthesis and isolation.
terpenoide; cucurbitacine and its biosynthesis and isolation purification and characterization
Metabolomic Profiling of Spent Biomass Of Marine Microalgae, Chlorella vulgarispriyanka raviraj
OBJECTIVE:
To evaluate the presence of any high value added compounds in the spent biomass of C. vulgaris
To identify the biological activity of the extracted compounds
To evaluate the structure and nature of the compounds using Nuclear Magnetic Resonance Spectroscopy and other analytical techniques.
Development of economically viable methodologies for the simultaneous extraction of by-products from a single set of biomass.
biological activities performed -Total antioxidant capacity, Anti bacterial activity, Anti-tuberculosis activity, Anti proliferative assay
LC-MS/MS method for the quantification of carbinoxamine in human plasmaIOSR Journals
A simple, reverse-phase high performance liquid chromatographic method with mass spectrometric detection (HPLC-MS/MS) was developed for determination of carbinoxamine in human plasma using pargeverine HCl as an internal standard. The procedure involves a simple protein precipitation technique using BDS HYPERSIL C8 (100 x 4.6mm) column. The mobile phase used was acetonitrile: buffer (25mm ammonium formate solution) (80:20). Precipitation was done using acetonitrile and detection was done in MRM mode, using an Electro Spray positive ionization. The ion transition monitored was (m/z) carbinoxamine (Q1 Mass: 291.2; Q3 Mass: 167.1), Internal standard (Q1 Mass: 338.1; Q3 Mass; 167.0). The retention time of carbinoxamine and internal Standard were 1.61 and 1.75 respectively. Method was evaluated in terms of linearity, accuracy, precision, recovery, sensitivity. The simple extraction procedure and short chromatographic runtime make the method suitable for therapeutic drug monitoring studies.
Research Inventy : International Journal of Engineering and Scienceresearchinventy
Research Inventy : International Journal of Engineering and Science is published by the group of young academic and industrial researchers with 12 Issues per year. It is an online as well as print version open access journal that provides rapid publication (monthly) of articles in all areas of the subject such as: civil, mechanical, chemical, electronic and computer engineering as well as production and information technology. The Journal welcomes the submission of manuscripts that meet the general criteria of significance and scientific excellence. Papers will be published by rapid process within 20 days after acceptance and peer review process takes only 7 days. All articles published in Research Inventy will be peer-reviewed.
DEVELOPING CRYO-ELECTRON MICROSCOPY OF BIOMOLECULES IN WATERGuttiPavan
Cryo-electron microscopy (Cryo-EM) is a type of transmission electron microscopy that allows for the specimen of interest to be viewed at cryogenic temperatures (-150°C)
Following years of improvement, the cryo-electron microscope has become a valuable tool for viewing and studying the 3D structures of various biological molecules in water.
Experimental Designs in Next Generation Sequencing GuttiPavan
Experimental Designs in Next Generation Sequencing
Introduction
Types of experimental designs
Basic NGS chemistry
Tools used in NGS
Good and Bad experimental designs
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
Ultrasonication Assisted Extraction of Isolation Characterization of Berberine from Annamita Cooculs Stem & it’s biological activities
1. PROJECT TITLE
Ultrasonication Assisted Extraction of Isolation
Characterization of Berberine from Annamita
Cooculs Stem & it’s biological activities.
Submitted By
GUTTI . PAVAN
M.Sc -2 , Biochemistry
Under The Guidance of
Dr. UTKARSHA LEKHAK
Department of Biochemistry,
The Institute of Science Mumbai 400032
2. SN TOPIC SN TOPIC
1 Aims 10 Determination of berberine content
2 Phytochemicals introduction 11 Colum chromatography
3 Alkaloids 12 Spectra analysis of berberine
4 Berberine 13 Antioxidant studies
5 Anamirta cocculus 14 Anti inflammatory studies
6 Project overview 15 Antimicrobial studies
7 Preparation stem water extract 16 Conclusion
8 Phytochemical ( alokilds) qualitative analysis : 17 Acknowledgement
9 Determination of alkaloid by BCG method 18 Bibliography
INDEX
3. To detect the presence of alkaloids in extracts by qualitative
method.
Isolation and characterization of berberine.
To detect antioxidant & anti-inflammatory activity in plant
extracts.
Antimicrobial activity studies.
AIMS
4. PHYTOCHEMICALS INTRODUCTION
Phytochemical is a broad term meaning plant (photo) chemical
referring to a wide variety of biologically activity compounds that
occur naturally in plants. which produced health benefits for
humans as medicinal ingredients and nutrients .
5. Alkaloids
Alkaloids are natural
product that contains
heterocyclic nitrogen
atoms, are basic in
character.
Alkaloids are naturally
synthesis by a large
numbers of organisms,
including animals, plants,
bacteria and fungi
6. BERBERINE
Berberine is a quaternary ammonium salt from
the protoberber.ine group of benzylisoquinoline alkaloids.
Berberine is an alkaloid found in a wide variety of traditional
plants.
Berberine has a broad spectrum of pharmacological activities.
Berberine comes to us from India and China and, where it was
first used in traditional Chinese medicine and Ayurvedic medicine.
Berberine compound is supplemented for its anti-diabetic and
anti-inflammatory effects.
7. anamirta cocculus
Anamirta cocculus (Marathi: काकमारी) is a Southeast
Asian and Indian climbing plant. Its fruit is the source
of picrotoxin, a poisonous compound with stimulant
properties.
Chemical substances
• The stem and the roots contain quaternary alkaloids,
suchas berberine, palmatine, magnoflorine and columba
mine.
• The seeds deliver picrotoxin, a sesquiterpene, while the
seed shells contain the tertiary
alkaloids menispermine and paramenispermine.
9. Preparation stem water
extract
Weight 2 grams of stem powder
with 50 ml distilled water in 250 ml
standard conical flask. mix well
properly. then keep the water flask in
sonication, at 37 c, for 30 mins.
The crude aqueous extract was
acidified with 1N HCl and then
treated with 1N NaOH. The aqueous
layer was further extracted using
chloroform.
The extraction was repeated until
the last fraction did not give any
precipitate (orange color) with
Dragendorff’s reagent.
10. Phytochemical ( alokilds) Qualitative Analysis :
1. Dragondroff’s reagent test
• take 2 ml plant extract into the test tube and add 5 drops dragendorff reagent, The formation of an
orange-red colored precipitate indicated the presence of alkaloids.
2. Mayer's test
• take 2 ml plant extract into the test tube and add 5 drops Mayer's reagent reagent, The formation of an
cream precipitate indicate the presence of alkaloids.
3. Hager test
• take 2 ml plant extract into the test tube and add 5 drops hangers reagent, The formation of a yellow
precipitate indicate the presence of alkaloids.
4.wagner test
• take 2 ml plant extract into the test tube and add 5 drops Wagner reagent, The formation of a brown coal
precipitate indicates the presence of alkaloids.
5.scheibler test
• take 2 ml plant extract into the test tube and add 5 drops scheibler reagent, The formation of a light yellow
-white precipitate indicates the presence of alkaloids.
6.sonnensceins test
• take 2 ml plant extract into the test tube and add 5 drops sonnensceins reagent, The formation of a light
yellow precipitate indicates the presence of alkaloids.
11. A-DRAGENDORFF TEST : orange -red color ppt , B- HAGERS TEST: Yellow color PPT ,
C- MAYER TEST: Cream color ppt , D- WAGER TEST: Red- brown colored PPT ,
E-SCHEUBLER TEST: white with light yellow , F- SONNENSCEINS TEST: Light yellow color
13. Determination of berberine content
• Take 1 ml plant extract
sample The absorbance for
test and standard solutions
were determined against
the blank ( only methanol )
at 420 nm .
• berberine sulphate Was
used as the standard
concentration of (40, 80,
120 ,160 and 200 μg/ml) .
14. colum chromatography
SAMPLE : load 1 ml crude sample ( 53
mg/ml).
SOLVENT SYSTEM : when finished the
loading sample into column, after runs with
column using different concentrations of
solvent system CHCl3: MeOH (in ratios of
98:2, 96:4, 95:5, 90:10, 80:20 and 70:30) .
SUCCESSIVE SOLVENT SYSTEM : The
CHCl3: MeOH (98:2) fraction yielded a
single peak in TLC, which was identified as
berberine by UV-Visible spectroscopy, FT-IR
and 1H NMR and LCMS.
16. UV data
The UV spectrum of
isolated berberine
sample and standard
berberine was found to
overlap, indicating the
purity of the spots.
The absorption
spectrum of berberine
in methanol solution is
424 nm.
17. IR DATA
FTIR spectra were recorded on
Shimadzu IR-Spectrometer (FTIR 8201)
central university of Hyderabad, at
room temperature within the wave
number range of 4000 to 400 cm−1
using KBr discs. The observed IR
absorption peak at ranges of
3500 cm−1 represents the N-H stretch
group, peak 2951.80cm−1 indicates
to C-H stretch, peak 1253cm−1 C-H
stretch aromatic, 2731 cm−1 shows C-
H stretch alkane, 1622 cm−1 peak
shows C-C stretch ring aromatic , peak
1328 cm−1 shows C-N stretch ,
mainly 1597 cm−1 shows benzene ,
1564 cm−1 indicates C=N, 972 cm−1
peak indicates O-C-O groups.
18. H1 NMR DATA
• 1H NMR spectra were recorded
in CDCl3. All assignments of the
proton atoms were found in
their expected region.
• The NMR spectra of berberine
was confirmed the absence of in
the berberine the signals
corresponding to two CH3
groups is observed at 1.68 and
1.64 ppm, (6H) C-H protons at
7.49 , 7.42, 6.83, 7.28, 7.24 and
6.85 ppm , (3H) CH2 protons at
1.64, 2.19 and 2.03 ppm.
19. Mass spectra data
Mass spectra provide an
essential clue for elucidating the
structure of compounds.
The esi mass spectra of the
berberin were recorded and
used to compare their
stoichiometry composition. The
berberin displays the prominent
molecular ion peak (m+) at 𝑚/𝑧
=336.1235
21. ANTI INFLAMMATORY ACTIVITY
• .
Procedure The anti-inflammatory
activity was measured by the method
of (standard protocol, Plant Research
Scheme).
Inflammation is a complex process,
which is frequently associated with
pain and involves occurrences
such as: the increase of vascular
permeability, increase of protein
denaturation and membrane
alteration.
When cells in the body are
damaged by microbes, physical
agents or chemical agents, the
injury is in the form stress.
Inflammation of tissue is due to
response to stress
22. DPPH Radical Scavenging Activity
Procedure ferric reducing antioxidant
(FRAP)The activity was measured by the
method of (KalaisezhiyenPavithra, 2015.
Take different volume of plant(
5,10,20,40,80.100,200 and 300 ug/ml) extracts
were made up to 40 ul with DMSO and 2.96 ml
DPPH (0.1 mM) solution was added. The
reaction mixture was incubated in dark
condition at room temperature at 20 mins,
after 20 mins the absorbance of the mixture
was measured at 517 nm .
2ml of DPPH was taken as control. Prepare one
blank solution which contains the ml of 9 ml
reagent and 1ml 1 ml distilled water.
The following equation was used to determine
the percentage of the radical scavenging
activity of each extract.
Percentage of radical scavenging activity=[(OD
control- OD sample)/OD control] × 100
23. The antioxidant activity of samples was evaluated by
the green phosphomolybdenum complex formation
according to the method of Prieto(1999).
Ferric reducing antioxidant
power assay Phosphomolybdenum assay
Procedure ferric reducing antioxidant (FRAP)The
activity was measured by the method of (standard
protocol, Plant Research Scheme)
24. ABTS activity aassay:
• ABTS radical scavenging activity was
estimated by following re et.al(1999)
method.
Hydroxyl radical scavenging activity
Hydroxyl radical scavenging activity of the
extracts was determined according to the
method reported by (Klein et al).
25. The chelating ability of the extract against iron was
determined using the method of Puntel et al.
Reducing power assay
• Reducing the power of MPE was
determined according to the
developed method
Hydrogen peroxide scavenging assay
26. Iron Chelation Assay
• The chelating ability of the extract against iron was determined using the method of
Puntel et al.
28. Steps
1.Collection of bacterial strains
• Pure cultures of all experimental bacteria were obtained from the
microbiology department of the Institute of science Mumbai,
Maharashtra. The pure bacterial cultures were maintained on nutrient
agar medium.
2.Bacterial Inoculums preparation
• Make The different bacterial suspensions( at lest 1ml) were adjusted
with sterile saline to a concentration of 1.0 X 108 CFU/ml. Inoculums
preparation Each bacterial strain was subcultured overnight at 35 C in
Mueller-Hilton agar slants. The bacterial growth was harvested using 5
ml of sterile saline water, its absorbance was adjusted at 580 um and
diluted to attain viable cell count of 108 CFU/ml using
spectrophotometer.
29. 3.Media Preparation and Its Sterilization:
• Antimicrobial susceptibility was tested on solid Agar-agar media (gm/100
ml: peptone 1gm , NaCl 0.5gms, yeast extract 0.3 grams and agar 4.gms
maintain pH 7.1) was used for developing surface colony growth. take above
chemical into starization conical flask boiled to dissolve the contents of the
medium and after pack with sterilization cotton. It is sterilized by
autoclaving at 121ºC for 20 minutes at 15 Lbs. pressure.
4.Pour media into a petri dish, and sample loading
• take 20 ml media into which were already washed and sterilized petri dish.
Do under near flame and Petri dishes( contain media ) keep for turn to
semi-solid phase under sterilization conditions. and loaded one control disc
(chloramphenicol (30 µg/disc) , respectively. The plates were incubated for
24 h at 37 °C.
5.Measurement zone of inhibition
• The measured diameters of the zone of inhibition for berberine against
different bacteria were measured in millimetre by using scale .recorded and
considered an an indication for antibacterial activity.
34. CONCLUSION
The berberine alkaloid was first time isolated at 370 C from stem Anamirta
cocculus plant using ultrasonication extraction method. The CHCl3: MeOH (98:2)
fraction yielded a single peak in TLC, which was identified as berberine by UV-
Visible spectroscopy. After the extraction of Berberine for the confirmation, TLC
was also performed to confirm the Rf value to be 0.52.
The experimental values of IR, UV , MS and H1NMR are used to predict the
molecular structure, atomic stretching, possible molecular functional group, etc.,
for the confirmation of berberine alkaloid present in the Anamirta cocculus plant
and are confirmed. The theoretical values are good agreement with the
experimental values.
The molecular structure is found by Gaussian software (V3) and also IUPAC name
by Chem Doodle software and is represented as : 5,6-dihydro-9,10-
dimethoxybenzo[g]-1,3-benzodioxolo[5,6a]quinolizinium). and the molecular
formula is C20H18NO.
35. The Anamitra cooclus water stem extract has great potential as
antimicrobial compounds against microorganisms. Thus, they can
help to use in the treatment of infectious diseases in human and
fruit spoilage caused by resistant microbes.
Food spoilage is often caused by the growth of many different
pathogenic bacterial strains. Prevention of food spoilage in the food
industry and foodstuff is mainly based on the application of
chemical preservatives.
we concluded The anamirta cocculus plant water extracts which
proved to be potentially effective as (anamirta cocculus ) can be
used as natural alternative preventives to control food poisoning
diseases and preserve foodstuff avoiding healthy hazards of
chemically antimicrobial agent applications.
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38. ACKNOWLEDGEMENT
• I would like to express my special thanks of gratitude to my Guide
Hon’ble Dr.Utkarsha leahak, who gave me the golden opportunity
to do this wonderful PPT “Ultrasonication Assisted Extraction of
Isolation Characterization of Berberine from Annamita Cooculs
Stem & it’s biological activities.” for her constant support and
motivation that has encouraged me to come up with the
assignment. Secondly i would also like to thank my Parents and
Friends (M.Sc-2 Biochemistry IOS) and Non-teaching staff who
have rendered their whole hearted support to all times for the
successful completion of this assignment .
My Family
Ios Biochemistry non-teaching staff
My M.Sc Biochemistry friends
Hon’ble
Dr.Utkarsha leahak