Isolation and Purification of Secoisolariciresinoldiglucoside oligomers (Lign...IOSR Journals
The present study aimed to extract and purify the compound of Secoisolariciresinoldiglucoside
oligomers (lignan) from flax seed (Linumusitatissimum) and its antioxidant activity. The Lignan was extracted
by solvents which gave the best results were ethanol : 1,4 dioxane (1:1, v:v).SDG release after alkaline
hydrolysisby using a methanolicNaOH , 20 mM, pH=8 at 50 ºC.followed by using following chromatographic
techniques: Liquid-liquid, Sephadex LH-20 column chromatography, thin layer chromatographic (TLC), high
performance liquid chromatographic (HPLC) and Fourier Transform Infra-Red(FTIR) . The EC50 values of
Pure lignan extract (9 μg/ml) was shown possess DPPH radical scavenging activity compared to reference
substances BHT and vitamin C (EC50= 3 and 4.2 μg/ml) respectively, and this was higher than partial pure
lignan component (EC50= 25.5 μg/ml).The total phenolic content of the pure lignanwas higher than partial
pure lignan which gave 22.312 and 14.85 g/ml respectively.
Separation of L-Phenylalanine by Solvent Sublation and Solvent Extraction MethodBRNSS Publication Hub
Aims and Objectives: Separation and purification is a series of processes intended to isolate a single type of biomolecule from a complex mixture. Innovations in improvement of biodownstream processing, which is responsible for the separation of about 50–80% of recombinant proteins and other biomolecules, play a very important role in increasing the yield and reducing the cost of biopharmaceutical production. Methods: Biomolecule isolation and purification from a fermentation broth usually involve centrifugation, filtration, adsorption, and chromatography steps. Results and Discussions: Each step contributes to the product cost and product loss. Thus, we consider that solvent extraction and solvent sublation are the more economic processes for the separation of biomolecules. Conclusion: In extraction of phenylalanine, maximum extraction was observed at amino acid: surfactant ratio 1:1, amino acid: extractant ratio 1:1500, and pH at 3.1. The highest value of % recovery percentage and Co/Cw was 76.3 and 10.21, respectively. The main motive of this article is to provide the advantages of study on the solvent sublation and solvent extraction of l-phenylalanine over the other techniques.
Extraction, identification and antioxidant activities of carotenoids from Ipo...Pragati Shah
These presentation slides are about extraction, identification and antioxidant activities of carotenoids from Ipomoea aquatica Forsk. It is commonly known as Morning water glory or water spinach which is rich source of nutrients,, wide array of carotenoids are extracted and identified by GC- MS techniques by researcher arond the Globe. Antioxidant activity of natural products can be evaluated by different assay; some them are discussed in these slides.
Isolation and Purification of Secoisolariciresinoldiglucoside oligomers (Lign...IOSR Journals
The present study aimed to extract and purify the compound of Secoisolariciresinoldiglucoside
oligomers (lignan) from flax seed (Linumusitatissimum) and its antioxidant activity. The Lignan was extracted
by solvents which gave the best results were ethanol : 1,4 dioxane (1:1, v:v).SDG release after alkaline
hydrolysisby using a methanolicNaOH , 20 mM, pH=8 at 50 ºC.followed by using following chromatographic
techniques: Liquid-liquid, Sephadex LH-20 column chromatography, thin layer chromatographic (TLC), high
performance liquid chromatographic (HPLC) and Fourier Transform Infra-Red(FTIR) . The EC50 values of
Pure lignan extract (9 μg/ml) was shown possess DPPH radical scavenging activity compared to reference
substances BHT and vitamin C (EC50= 3 and 4.2 μg/ml) respectively, and this was higher than partial pure
lignan component (EC50= 25.5 μg/ml).The total phenolic content of the pure lignanwas higher than partial
pure lignan which gave 22.312 and 14.85 g/ml respectively.
Separation of L-Phenylalanine by Solvent Sublation and Solvent Extraction MethodBRNSS Publication Hub
Aims and Objectives: Separation and purification is a series of processes intended to isolate a single type of biomolecule from a complex mixture. Innovations in improvement of biodownstream processing, which is responsible for the separation of about 50–80% of recombinant proteins and other biomolecules, play a very important role in increasing the yield and reducing the cost of biopharmaceutical production. Methods: Biomolecule isolation and purification from a fermentation broth usually involve centrifugation, filtration, adsorption, and chromatography steps. Results and Discussions: Each step contributes to the product cost and product loss. Thus, we consider that solvent extraction and solvent sublation are the more economic processes for the separation of biomolecules. Conclusion: In extraction of phenylalanine, maximum extraction was observed at amino acid: surfactant ratio 1:1, amino acid: extractant ratio 1:1500, and pH at 3.1. The highest value of % recovery percentage and Co/Cw was 76.3 and 10.21, respectively. The main motive of this article is to provide the advantages of study on the solvent sublation and solvent extraction of l-phenylalanine over the other techniques.
Extraction, identification and antioxidant activities of carotenoids from Ipo...Pragati Shah
These presentation slides are about extraction, identification and antioxidant activities of carotenoids from Ipomoea aquatica Forsk. It is commonly known as Morning water glory or water spinach which is rich source of nutrients,, wide array of carotenoids are extracted and identified by GC- MS techniques by researcher arond the Globe. Antioxidant activity of natural products can be evaluated by different assay; some them are discussed in these slides.
Extraction and Isolation of Lycopene Form Various Natural Sourcesiosrjce
Lycopene is a carotenoid pigment and phytochemical found in tomatoes, water melon and other
fruits mostly red coloured. The antioxidant properties of lycopene have attracted attentions due to its biological
properties and are thought to be primarily responsible for its benefits in health related problems. Extraction
and isolation of lycopene can be done with various methods. Several studies have been undertaken for the
extraction of lycopene and for its quantification in various natural sources.
Lipid profiling and corresponding biodiesel quality of mortierella isabellina...zhenhua82
Four lipid extraction methods (Bligh & Dyer, hexane & isopropanol, dichloromethane & methanol, and hexane) were evaluated to extract lipid from freeze- and oven-dried fungus Mortierella isabellina ATCC42613. The highest lipid yield (41.8%) was obtained from Bligh & Dyer extraction on the oven-dried fungal biomass with a methanol:chloroform:water ratio of 2:1:0.8. Other lipid extraction methods on both freeze- and oven-dried samples had lipid yields ranging from 20.7% to 35.9%. Non-polar lipid was the main lipid class (more than 90% of total lipid) in M. isabellina. Regarding fatty acid profile, there was no significant difference on fatty acid concentration between different drying and extraction methods. Estimation of biodiesel fuel properties using correlative models further demonstrated that the fungal biodiesel is a good alternative to fossil diesel.
Metabolic Pathways in Higher Plants and their DeterminationDr. Siddhi Upadhyay
a) Brief study of basic metabolic pathways and formation of different secondary metabolites through these pathways- Shikimic acid pathway, Acetate pathways and Amino acid pathway.
b) Study of utilization of radioactive isotopes in the investigation of Biogenetic studies.
Flavonoids classification, isolation and identificationMona Ismail
Flavonoids are groups of polyphenolic compounds which are found in fruits, flowers, seeds & vegetable.
(named from the Latin word flavus meaning yellow, their colour in nature)
Chemistry of Natural Products
Alkaloids
• Introduction; classification; isolation; general methods for structure elucidation; discussion with particular reference to structure and synthesis of ephedrine, nicotine, atropine, quinine, papaverine and morphine.
• Terpenoids
• Introduction; classification; isolation; general methods for structure elucidation; discussion with particular reference to structure and synthesis of citral, α-terpineol, α-pinene, camphor and α-cadinene.
• Steroids
• Introduction; nomenclature and stereochemistry of steroids; structure determination of cholesterol and bile acids; introduction to steroidal hormones with particular reference to adrenal cortical hormones.
Extraction and Isolation of Lycopene Form Various Natural Sourcesiosrjce
Lycopene is a carotenoid pigment and phytochemical found in tomatoes, water melon and other
fruits mostly red coloured. The antioxidant properties of lycopene have attracted attentions due to its biological
properties and are thought to be primarily responsible for its benefits in health related problems. Extraction
and isolation of lycopene can be done with various methods. Several studies have been undertaken for the
extraction of lycopene and for its quantification in various natural sources.
Lipid profiling and corresponding biodiesel quality of mortierella isabellina...zhenhua82
Four lipid extraction methods (Bligh & Dyer, hexane & isopropanol, dichloromethane & methanol, and hexane) were evaluated to extract lipid from freeze- and oven-dried fungus Mortierella isabellina ATCC42613. The highest lipid yield (41.8%) was obtained from Bligh & Dyer extraction on the oven-dried fungal biomass with a methanol:chloroform:water ratio of 2:1:0.8. Other lipid extraction methods on both freeze- and oven-dried samples had lipid yields ranging from 20.7% to 35.9%. Non-polar lipid was the main lipid class (more than 90% of total lipid) in M. isabellina. Regarding fatty acid profile, there was no significant difference on fatty acid concentration between different drying and extraction methods. Estimation of biodiesel fuel properties using correlative models further demonstrated that the fungal biodiesel is a good alternative to fossil diesel.
Metabolic Pathways in Higher Plants and their DeterminationDr. Siddhi Upadhyay
a) Brief study of basic metabolic pathways and formation of different secondary metabolites through these pathways- Shikimic acid pathway, Acetate pathways and Amino acid pathway.
b) Study of utilization of radioactive isotopes in the investigation of Biogenetic studies.
Flavonoids classification, isolation and identificationMona Ismail
Flavonoids are groups of polyphenolic compounds which are found in fruits, flowers, seeds & vegetable.
(named from the Latin word flavus meaning yellow, their colour in nature)
Chemistry of Natural Products
Alkaloids
• Introduction; classification; isolation; general methods for structure elucidation; discussion with particular reference to structure and synthesis of ephedrine, nicotine, atropine, quinine, papaverine and morphine.
• Terpenoids
• Introduction; classification; isolation; general methods for structure elucidation; discussion with particular reference to structure and synthesis of citral, α-terpineol, α-pinene, camphor and α-cadinene.
• Steroids
• Introduction; nomenclature and stereochemistry of steroids; structure determination of cholesterol and bile acids; introduction to steroidal hormones with particular reference to adrenal cortical hormones.
IOSR Journal of Applied Chemistry (IOSR-JAC) is an open access international journal that provides rapid publication (within a month) of articles in all areas of applied chemistry and its applications. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Chemical Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Phytochemical and Biological Evaluation of Cassia tora, L. Seedsiosrjce
In the present study the total and the defatted alcoholic extracts of seeds of Cassia tora Linn.
(Leguminosae) were screened for hepatoprotective activity using adult Wister albino rats (120-170 g) as the
experimental animals. Hepatic injury caused by carbon tetra chloride, was analyzed through estimation of AST,
ALT, ALB and platelets in blood samples taken from the veins of orbital plexus of each animal as well as the
histopathological examination of the liver.The effects of the extracts were comparable with standardhepatoprotective drug Silymarin. On the other hand GC-MS analysis was performed on the fatty acid
composition of the lipoidal fraction for the seeds. The separated fatty acids were converted to their methyl ester
and then subjected to the analysis.
Degradation of an organophosphorus insecticide (chlorpyrifos) in simulated wa...Salah Hussein
Induced degradation of chlorpyrifos insecticide in simulated wastewater with advanced oxidation processes (AOPs), using ultraviolet irradiation (UV), ozonation and chemical oxidation using (sodium hypochlorite, calcium hypochlorite, monochloride-isocyanuric acid (MCICA), dichloroiso-cyanuric acid (DCICA), trichloroisocyanuric acid (TCICA) ) was studied. Chlorpyrifos and its degradation products were extracted using solid phase extraction (SPE) method, identified using GC-MS. Results showed that the degradation of chlorpyrifos in simulated wastewater followed the first order reaction, and its half life was 3.34, 5.64, 7.13 and 10.69h under ozonation, UV, 1.5%TCICA and 1.5%DCICA respectively when chlorpyrifos solutions treated for 12 h. The concentrations of chemical oxidative substances, active chlorine content and time of treatments had a significant effect on degradation rate of chlorpyrifos, which increased with increasing of each. The most enhancement of chlorpyrifos degradation was observed in treatment with ozonation, UV, TCICA and DCICA where the dissipations % of the parent compounds were 85.70, 57.71, 43.71 and 35.07 %, respectively. The intermediates products of chlorpyrifos degradation using chemical method were identified as O,O-Diethyl thiophosphate(DEP), 3,5,6-trichloro-2-pyridinol(TCP), 3,5,6-trichloro-2-methoxypyridine(TMP) and 2,3,5,6-tetrachloro-pyridine. UV leads to formation of O,O-Diethyl phosphate, TCP and Chlorpyrifos oxon. Ozonation leads to formation of O,O-Diethyl thiophosphate beside the UV degradation products.
Total phenolics and total flavonoids of extracts from freshwater Clam (Corbic...Innspub Net
The ethanol, ethyl acetate, and hexane extracts of the freshwater clam (Corbicula fluminea) were studied for the total phenolics and total flavonoids. Total phenolics and total flavonoids of the extracts were evaluated using Folin-Ciocalteau and Aluminum chloride colorimetric methods respectively. The findings showed that the total phenolics of the ethanol extract (1.67±0.28mg GAE/g of dried sample) were substantially higher than the total phenolics obtained from the ethyl acetate (0.70±0.00mg GAE/g) and hexane extracts (0.56±0.23mg GAE/g). While the total flavonoids in the ethyl acetate extract displayed a slightly higher total flavonoid (43.84±0.92mg QE/g of dried sample) relative to ethanol (30.41±1.34mg QE/g of dried sample) and hexane extracts (20.28±0.00mg QE/g of dried sample). Using ethanol, the highest yield for extraction was obtained. Ethanol is the best solvent among the three – ethanol, ethyl acetate, and hexane in terms of extraction yield and total phenolics. In addition, it can be inferred that the presence of significant amounts of phenolics and flavonoids suggests that freshwater clam is a promising source of antioxidants that provides nourishing proteins and oxidative stress remedies
A STUDY ON FORMATION OF SALYCILIC ACID FORMALDEHYDE POLYMER SAMPLEEDITOR IJCRCPS
Condensation of salicylic acid (0.02 mole) with formaldehyde (0.016 mole) in presence of aqueous 40% H2SO4.
Keywords: pipette,thermometer,spectro-photometer,conicalflakk,waterbath.
This is a study undertaken on nutrient compositions of and processed products development from Son Tra (Docynia indica. The in-depth study of some bioactive substances of son tra fruit and its processed product development was undertaken as part of understanding existing market value chains, and identifying the necessary interventions to improve the performance of son tra.
Biochemical components of three marine macroalgae (Padina pavonica, Ulva lact...Innspub Net
Green macroalgae Ulva lactuca, brown macroalgae Taonia atomaria and Padina pavonica are spread in the Turkish Levantine Sea. There is limited information about antioxidant activities and fatty acid composition of these species from Levantine Sea. In this study was to determine and compare antioxidant activities, vitamin and fatty acid (FA) composition of U. lactuca, T. atomaria and P. pavonica. The analysis was made with HPLC and GC device. g. Then, the results were analyzed using SPSS software. The results showed; palmitic acid (C16:0) as the most abundant saturate fatty acid (21-41%). The green algae was rich palmitic acid (C16:0) (41.68%). Monounsaturated fatty acids (MUFAs) were major components (39.81–42.89%). The total MUFA content for U. lactuca was 40.63%, P. pavonica 42.89% and for T. atomaria 38.81%. Oleic acid (C18:1 n-9) was the most abundant MUFA in all the species analyzed. Eicosapentaenoic acid ( C20:5 n-3) and arahidonic acid (C20:4 n-6) were found in significant levels in T. atomaria. P. pavonica and T. atomaria showed similar amounts of C18 and C20 PUFAs contents. In T. atomaria eicosopentaenoic acid (EPA, C20:5n3) accounted 4.78% of total fatty acids. PUFA/SFA ratio in T. atomaria was 1.10%, U. lactuca; 0.26% and for P. pavonica 0.68%.The total phenolic contents ranged from 0.96 to 2.22 mg gallic acid equivalents per 1 g of dry macroalgae material. Phenolic content of the water extract of T. atomaria (2.22 mg GAE /g) was higher than that of the water extract of P. pavonica and U. lactuca. It has been thought that the amount of α-tocoferol was higher than the other lipophilic vitamins in all the three species tested. In Conclusion; these species can be used as food and in food industry.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
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.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
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.
In silico drugs analogue design: novobiocin analogues.pptx
H0434555
1. IOSR Journal of Applied Chemistry (IOSR-JAC)
e-ISSN: 2278-5736.Volume 4, Issue 3 (May. – Jun. 2013), PP 45-55
www.iosrjournals.org
www.iosrjournals.org 45 | Page
Isolation And Identification Of Some Chemical Constituents In
Two Different Types Of Fresh Water Macro Algae In Bestansur
Village In Suleiman City Kurdistan Region (North Iraq) By Hplc
Technique
Trifa Attar Omer
Department of Chemistry, School of science, Faculty of science & science education, University of Suleiman.
Abstract: In the present work HPLC analysis for two genus of green algae were collected locally and identified
from Bestansur village fresh water in Sulaimani city in north Iraq in October 2011,shows that contain different
alkaloid components such as Chara sp. Contain (Kaemfertin 57.0831µgm/ml ,isoquinoline 74.8601µ gm/ml
,corilagin 109.3617µ gm/ml and colocynthitin 57.3929 µ gm/ml) while Spirogyra contain (Gallic acid 77.2055
µ gm/ml ,Kaempfertin 67.8407 µ gm/ml ,isoquinoline 69.6164 µ gm/ml , corilagin 105.8581 µ gm/ml and
colocynthitin 85.9098 µ gm/ml).
Divert compounds of steroids composition were also analyzed by HPLC technique in two types of
macro algae five steroid compounds were detected (stigmasterol 69.9213 µ gm/ml , 73.102 µ gm/ml ,
campsterol 98.8401 µ gm /ml ,157.8413 µ gm/ml , β-sitosterol 127.8065 µ gm/ml,37.1229 µ gm/ml ,avenasterol
43.15194 µ gm/ml ,72.400 µ gm/ml and clerosterol 89.6958 µ gm/ml ,84.50799 µ gm/ml respectively).
In this study HPLC analysis for Chara show that contain( α-tocopherol 334.53417 µ gm/ml , β-
tocopherol 175.13087 µ gm/ml and γ-tocopherol 71.7641 µ gm /ml) .
In the end of this study ICP analysis for two genus macroalgae shows that contain this element s (Mn,V,Se,
Ca,Mg, Cr,As,Pb,Co,Cd and Ni).
Key Words: HPLC analysis, macro algae ,Alkaloid, steroid, tocopherol
I. Introduction
Macro algae are major contributors to the total marine plant biomass (1).Currently, Macro algae are attracting
increasing interest ,in view of their low caloric content and high vitamin ,mineral and dietary fiber contents (2),
making them attractive to both consumers and the food industry (3).Numerous biological amines and
halogenated cyclic nitrogen –containing substances are included in the term alkaloid. The latter is specific from
marine organisms and marine algae. They could not be found in terrestrial plants .after the isolation of alkaloids;
pure active compounds were used in therapy instead of plant extracts. Isolation of active compounds from plants
begin in 18th
century .Morphine was the first alkaloid extracted from a terrestrial plant in 1805 as reported by
Keppel mayer (4) and hordenine was the first alkaloid isolated from a marine algae in 1969.(5,6).
As a consequence of an increasing demand for biodiversity in the screening programs seeking
therapeutic drugs from natural products , there is now a greater interest in marine organisms , especially algae.(
7)Sterols are an important family of lipids, present in the majority of the cells .because of different routes of
synthesis ,sterols from plants, fungi and animals show marked differences.
Tasende (2000) confirmed that fatty acids and sterol of algal class, families and sometimes even
species are characteristics to those particular taxa and could be useful as chemotaxonomic (8).Clinical studies
have demonstrated that dietary intake of plant sterols ( as part of the normal diet ,or as a supplement ) may help
reduce blood cholesterol level.(9)Vitamin E is a generic term applied to the tocopherols and tocotrienols (which
show similar nutritional properties to α-tocopherol (10) ,these substances are highly stable to heat and acids ,and
unstable to alkalis ,UV light and oxygen ;in addition ,they are regarded on contact with rancid fats, lead and iron
(11).α-tocopherol is a liposoluble compounds that is capable of fixing free radicals via its phenol group ,and is
thus considered to play an antioxidant role in biological membranes , lipoproteins and fat deposits, controlling
or reducing lipid peroxidation .
In addition, it has been suggested that it helps protect against cancer induced by free –radical –
generating contaminants, such as ozone or nitrogen dioxide (12).
2. Isolation and identification of some chemical constituents in two different types of fresh water macro
www.iosrjournals.org 46 | Page
Clerosterol
Fig: (1) Structure of Steroid compounds
Isoquinoline
corilagin
Figure :( 2) structure of alkaloids compound
3. Isolation and identification of some chemical constituents in two different types of fresh water macro
www.iosrjournals.org 47 | Page
Fig: (3) tocopherol structure
II. Experimental
Two types of Macro algae (chara sp. ,spirogyra) were collected locally and identified from fresh water
in Bestansur village in Sulaimani city in Kurdistan region-north Iraq in October 2011.Algae samples were
cleaned at epiphytes and necrotic parts were removed .
Samples were rinsed with sterile water to remove any associated debris .samples was kept under
sunshade for 7 days .after drying the samples , it was ground thoroughly to powder form .
After a morphological and chemical characterization, the samples were prepared for determination of
different compounds of alkaloids, steroids and tocopherol and ICP –analysis.
A / RP- HPLC determination of alkaloids in chara sp. & spirogyra.
Extraction procedure for alkaloids compound.
The alcoholic extract of alkaloids were separated on FLC (Fast liquid chromatographic) column, 3um
particle size (50×4.6 mm I.D) chiral coloumn,mobile phase were 0.01 M phosphate buffer PH 8.2: acetonitrile
(60:40 v/v) detection UV set as 220 nm ,Flow rate 0.9 ml/min , the sequence of the eluted material of the
standard were as follow , each standard was 25µ g/ml.
Assay results for calibration standards was of the linear range of 1-100 µg/ml , the recovery rate of
%96.8 ,RSD of %1.8 (n=5)
The separation occurred on liquid –chromatography shimadzu 2010 LC equipped with binary delivery
pump model 2010 shmadzu , the eluted peaks were monitored by UV-VIS 2010 SPD spectrophotometer.
Preparation of sample
1 gm of the sample added in 250 ml glass beaker 250 ml of 0.25% acetic acid in ethanol has been added ,the
beaker was covered with glass watch for 4 hrs , the extract where filtered and concentrated to 100 ml using
rotary evaporator drop of %5 ammonium hydroxide were added to precipitation the alkaloids , the precipitated
were filtered and dissolve in 1ml of mobile phase ,then separated on HPLC under the same condition of
standard. Concentration for each sample were quantitatively determined by comparison the peak area of the
standard with that of the samples.
B / Extraction procedure for steroid compounds.
Active ingredient sterols:
The extract were separated on FLC (Fast liquid chromatography , columns, C-18,3 µm particle size
(50×4.6 mm ID) ,mobile phase 0.1% acetic acid in deionized water (solvent A) and acetonitrile (solvent B)
using linear gradient from 0-100% B in 10 minutes , detection UV set as 275 nm , flow rate 1.2 ml/min
temperature 30 c.The separation occurred on liquid – chromatography shimadzu 2010 LC equipped with binary
pump model 2010 shimadzu the eluted peaks were monitored by UV-VIS 2010 SPD spectrophotometer.
Extraction procedure:
1 gm sample has been dissolved in 5 ml of hot water for 2 hour , then using Ultrasonic bath for 20
minutes to get all extract dissolved in hot water 60 c degree , then the extract were filtered on filter paper no
1,0.5 mm to separate the fiber , 20 µgm from the sample injected on HPLC system .
The sequences of the eluted material of the standard were as follow, each standard was 25 µg/ml.
C/ RP- HPLC determination of tocopherol
Fast soluble tocopherols were separated on FLC (Fast liquid chromatographic ) column ,discovering
HSC -18 ,3 µgm particle size (50×4.6 nm I.D) column , Mobile phase : acetonitrile , detection :UV set at 290
nm ,Flow rate 1.0 ml/min ,injection :20µL .the sequence of the eluted standard was 50 µg /ml.
4. Isolation and identification of some chemical constituents in two different types of fresh water macro
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D / Determination of elements :
The sample 1 gm. weighed and adds 2 drops of nitric acid conc. And the volume was completed to 50
ml distilled water the solution was injected to OES – ICP Perkin Elmer 2100 for determine.
III. Calculation
Concentration of sample µg/ml = area of sample/area of standard ×conc. of standard ×dilution factor
IV. Result and discussion:
HPLC analysis of alkaloid compounds in two green genus Macro algae ( chara and spirogyra) are
represented in table (1-2) , from the result chara sp. Contain four types of alkaloids like (Kaempfertin ,
isoquinoline ,corilagin and colocynthitin) , While spirogyra contain five types of alkaloid compounds (Gallic
acid ,Kaempfertin ,isoquinoline , corilagin and colocynthitin) ,HPLC Chromatogram's of two types of macro
algae for alkaloids show in fig (4),(5) and (6)
The structure of some alkaloids compound shown in Fig -1-
Two green genus Macro algae contain (Kaempfertin, isoquinoline, corilagin and colocynthitin) while chara sp.
not contain Gallic acid.
Also HPLC analysis of steroid compounds in two green genus macro algae (chara sp. and spirogyra)
are represented in table (3-4) , from the result chara and spirogyra contain five types of steroid compounds like
(stigmasterol , campesterol , β-sito sterol , avenasterol and clerosterol ) from the result appear that chara contain
high amount of β-sitosterol 127.80656 µgm/ml while spirogyra contain high amount of campesterol.,in Fig -2-
show structure of all steroids compound,HPLC Chromatogram's of two types of macro algae for steroids show
in fig (7),(8) and (9) HPLC analysis for tocopherol in chara sp.are represented in table (5) show that contain α-
tocopherol 334.530417 µgm/ml , β-tocopherol 175.13087 µgm/ml and γ- tocopherol 71.764118 µgm/ml
.from the result appear that contain high amount of α-tocopherol 334.530417 µgm/ml in Fig -3- show the
chemical structure of α- tocopherol and low amount of γ- tocopherol 71.764118 µgm/ml .HPLC
Chromatogram's of two types of macro algae for tocotocopherol show in fig (10), (11)
In the end Elemental analysis have been done by Inductive couple plasma (ICP) technique (OES – ICP
Perkin Elmer 2100) for each types of Macro algae and the result cited in table (6) .from the result show that
contain (Mn , V, Se ,Ca ,Mg ,Cr ,As ,Pb ,Co,Cd ,Ni) from the results appear that contain high amount of Ca
,Mg . And (Co) does not contain in spirogyra sp.
Table -1- Alkaloid compounds in chara sp
no subjects Rt/minute area/standard df area/sample conce.µg/ml
conce. of
sample
1 Gallic acid 1.17 29925 3 0.0000
2 Kaempfertin 2.19 40060 3 30490 25 57.0831
3 isoquinoline 3.11 45597 3 45512 25 74.8602
4 corilagin 4.3 28021 3 40859 25 109.3617
5 colocynthitin 5.19 49689 3 38024 25 57.3930
Table (2): alkaloid compounds in spirogyra
no subjects Rt/minute area/standard df area/sample conce.µg/ml
conce. of
sample
1 Gallic acid 1.17 29925 3 30805 25 77.2055
2 Kaempfertin 2.19 40060 3 36236 25 67.8407
3 isoquinoline 3.11 45597 3 42324 25 69.6164
4 corilagin 4.3 28021 3 39550 25 105.8581
5 colocynthitin 5.19 49689 3 56917 25 85.9099
5. Isolation and identification of some chemical constituents in two different types of fresh water macro
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Table (3): steroid compounds in char a sp.
no subjects Rt/minute area/standard df area/sample conce.µg/ml
conce. of
sample
1 stigmasterol 0.99 21221 3 19784 25 69.9213
2 campesterol 2.15 21597 3 28462 25 98.8401
3 β-sitosterol 3 22973 3 39148 25 127.8066
4 avenasterol 3.92 24496 3 14094 25 43.1519
5 clerosterol 4.99 19878 3 23773 25 89.6959
Table (4): steroid compounds in spirogyra
no subjects Rt/minute area/standard df area/sample conce.µg/ml
conce. of
sample
1 stigmasterol 0.99 21221 3 20684 25 73.1021
2 campesterol 2.15 21597 3 45452 25 157.8414
3 β-sitosterol 3 22973 3 11371 25 37.1229
4 avenasterol 3.92 24496 3 23647 25 72.4006
5 clerosterol 4.99 19878 3 22398 25 84.5080
Table (5): tocopherol compounds in chara sp
no subjects Rt/minute area/standard df area/sample conce.µg/ml
conce. of
sample
1
α -
tocopherol
2.08 18612 3 41509 50 334.5342
2
β-
tocopherol
2.89 36869 3 43046 50 175.1309
3
γ-
tocopherol
3.82 24012 3 11488 50 71.7641
Table 6: ICP determination of elements
Conc.in ppm
Spirogyra
Conc.in ppm
Chara sp
Elements
4.5781.744Mn
0.5060.403V
0.1560.181Se
1673931Ca
106149Mg
0.0350.039Cr
0.2090.271As
0.1460.150Pb
Nil0.208Co
2.05810.86Cd
0.2312.251Ni
6. Isolation and identification of some chemical constituents in two different types of fresh water macro
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Fig (4): HPLC Chromatograms' of alkaloids compounds in chara sp.
Fig (5): HPLC Chromatograms' of alkaloids compounds in spirogra sp.
7. Isolation and identification of some chemical constituents in two different types of fresh water macro
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Fig(6): HPLC Chromatograms of a mixture of authentic standards of alkaloids compounds
Fig (7): HPLC Chromatograms' of steroids compounds in chara sp.
8. Isolation and identification of some chemical constituents in two different types of fresh water macro
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Fig (8): HPLC Chromatograms' of steroids compounds in spirogyra sp.
Fig(9): HPLC Chromatogram's of a mixture of authentic standards of steroids compounds
9. Isolation and identification of some chemical constituents in two different types of fresh water macro
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Fig (10):HPLC Chromatograms' of tocopherol compounds in chara sp
10. Isolation and identification of some chemical constituents in two different types of fresh water macro
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Fig (11): HPLC Chromatogram's of a mixture of authentic standards of tocopherol compounds
V. Conclusion :
Different compounds of alkaloid occur in relatively high concentration in two green genus Macro algae
that have taken from Bestansur village from sulaimani city Kurdistan region –Iraq.
Different compounds of steroid also occur in relatively high concentration in two genus Macro algae
from Bestansur like (stigmasterol ,campesterol ,β- sitosterol ,avenasterol and clerosterol ).
Physterol occur as organic compounds and essential constituents of cell membranes in all plant oils.
Recently increased interest in phytosterol lies in their potential to reduce plasma.
Different tocopherol occurs in relatively high concentration especially α-tocopherol.
And the amount of these elements like Ca,Mg ,Cr, Cd ,As are exist in these these Macro algae that useful .
This validated method could help to identify the level of numerous toxic and bioactive elements in
Macro algae.
Acknowledgement:
We are very much indebted to the department of chemistry, faculty of science and science education,
university of sulaimani for providing the facilities, encouragement and financial support during the investigation
.also thanks due to the University of Bagdad for obtaining HPLC technique for identification of the phytosterol
compounds and alkaloids and tocopherol in two types of macro algae.
11. Isolation and identification of some chemical constituents in two different types of fresh water macro
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