Carbohydrates are one of the three macronutrients in the human diet, along with protein and fat. These molecules contain carbon, hydrogen, and oxygen atoms. Carbohydrates play an important role in the human body. They act as an energy source, help control blood glucose and insulin metabolism, participate in cholesterol and triglyceride metabolism, and help with fermentation. The digestive tract begins to break down carbohydrates into glucose, which is used for energy, upon consumption. Any extra glucose in the bloodstream is stored in the liver and muscle tissue until further energy is needed. Carbohydrates is an umbrella term that encompasses sugar, fruits, vegetables, fibers, and legumes. While there are numerous divisions of carbohydrates, the human diet benefits mostly from a certain subset.
Estimation of reducing and non reducing sugarJasmineJuliet
Reducing sugar definition and example, non-reducing sugar definition and example, Estimation of reducing sugar by DNSA method, Estimation of total sugars by anthrone metod, Estimation of non-reducing sugar from amount of total sugars and reducing sugar, formula for estimation of non-reduci
Role of immobilized Enzymes in Food industryJasmineJuliet
Immobilization techniques, Immobilization techniques in food industry, Immobilized Enzymes, Need for immobilization, Role of immobilized Enzymes in Food Industry, Methods of immobilization, Production of lactose free milk, Production of High Fructose corn syrups, Production of Juice in industry level by Immobilized enzymes of Pectinase, Meat tenderization by immobilized Enzymes, Immobilized Amino acylase, immobilized glucose isomerase, immobilized pectinase, Immobilized alkaline phosphatase.
This ppt explains the properties of monosaccharides, polysaccharides. the properties like mutarotation, reduction, optical activity, caramerlization, osazone is given in the ppt. Also the determination of ring size of the monosaccharide is explained/
Estimation of reducing and non reducing sugarJasmineJuliet
Reducing sugar definition and example, non-reducing sugar definition and example, Estimation of reducing sugar by DNSA method, Estimation of total sugars by anthrone metod, Estimation of non-reducing sugar from amount of total sugars and reducing sugar, formula for estimation of non-reduci
Role of immobilized Enzymes in Food industryJasmineJuliet
Immobilization techniques, Immobilization techniques in food industry, Immobilized Enzymes, Need for immobilization, Role of immobilized Enzymes in Food Industry, Methods of immobilization, Production of lactose free milk, Production of High Fructose corn syrups, Production of Juice in industry level by Immobilized enzymes of Pectinase, Meat tenderization by immobilized Enzymes, Immobilized Amino acylase, immobilized glucose isomerase, immobilized pectinase, Immobilized alkaline phosphatase.
This ppt explains the properties of monosaccharides, polysaccharides. the properties like mutarotation, reduction, optical activity, caramerlization, osazone is given in the ppt. Also the determination of ring size of the monosaccharide is explained/
Introduction of fats, Reaction of fatty acids, Reaction of fats or oil- Hydrolysis, Hydrogenation, Halogenation, saponification, Drying of oil, Rancidity, Determination of acid value, saponification value, iodine value, acetyl value,
Fermentation in food processing is the process of converting carbohydrates to alcohol or organic acids using microorganisms—yeasts or bacteria under anaerobic conditions.
Or
Any metabolic process that releases energy from a sugar or other organic molecule, does not require oxygen or an electron transport system, and uses an organic molecule as the final electron acceptor
Fermentation usually implies that the action of microorganisms is desired.
The science of fermentation is known as zymology.
in microorganisms, fermentation is the primary means of producing ATP by the degradation of organic nutrients anaerobically
Rancidification is the process of complete or incomplete oxidation or hydrolysis of fats and oils when exposed to air, light, or moisture or by bacterial action, resulting in unpleasant taste and odor. Specifically, it is the hydrolysis or autoxidation of fats into short-chain aldehydes and ketones, which are objectionable in taste and odor. When these processes occur in food, undesirable odors and flavors can result.
CYTOLOGY 2
BIOCHEMISTRY
ORGANIC CONSTITUENT OF THE CELLS.
Bio chemistry: is the study of structures, properties and functions of chemical constituents of the cells.
-It is a great unifying theme in biology.
It finds applications in fields like;
1. Agriculture; in developing pesticides and herbicides.
2. Medicine; including all pharmaceuticals.
3. Fermentation; baking products, food products and breweries.
4. New development of biology eg genetic engineering.
ELEMENTS FOUND IN LIVING ORGANISMS ARE
Introduction of fats, Reaction of fatty acids, Reaction of fats or oil- Hydrolysis, Hydrogenation, Halogenation, saponification, Drying of oil, Rancidity, Determination of acid value, saponification value, iodine value, acetyl value,
Fermentation in food processing is the process of converting carbohydrates to alcohol or organic acids using microorganisms—yeasts or bacteria under anaerobic conditions.
Or
Any metabolic process that releases energy from a sugar or other organic molecule, does not require oxygen or an electron transport system, and uses an organic molecule as the final electron acceptor
Fermentation usually implies that the action of microorganisms is desired.
The science of fermentation is known as zymology.
in microorganisms, fermentation is the primary means of producing ATP by the degradation of organic nutrients anaerobically
Rancidification is the process of complete or incomplete oxidation or hydrolysis of fats and oils when exposed to air, light, or moisture or by bacterial action, resulting in unpleasant taste and odor. Specifically, it is the hydrolysis or autoxidation of fats into short-chain aldehydes and ketones, which are objectionable in taste and odor. When these processes occur in food, undesirable odors and flavors can result.
CYTOLOGY 2
BIOCHEMISTRY
ORGANIC CONSTITUENT OF THE CELLS.
Bio chemistry: is the study of structures, properties and functions of chemical constituents of the cells.
-It is a great unifying theme in biology.
It finds applications in fields like;
1. Agriculture; in developing pesticides and herbicides.
2. Medicine; including all pharmaceuticals.
3. Fermentation; baking products, food products and breweries.
4. New development of biology eg genetic engineering.
ELEMENTS FOUND IN LIVING ORGANISMS ARE
Structure and function of immunoglobulins(antibodies) Likhith KLIKHITHK1
Immunoglobulins (Ig) or antibodies are glycoproteins that are produced by plasma cells. B cells are instructed by specific immunogens.For, example, bacterial proteins, to differentiate into plasma cells, which are protein-making cells that participate in humoral immune responses against bacteria, viruses, fungi, parasites, cellular antigens, chemicals, and synthetic substances.
The immunogen or antigen reacts with a B-cell receptor (BCR) on the cell surface of B lymphocytes, and a signal is produced that directs the activation of transcription factors to stimulate the synthesis of antibodies, which are highly specific for the immunogen that stimulated the B cell. Furthermore, one clone of B cell makes an immunoglobulin (specificity). Besides, the immune system remembers the antigens that caused a previous reaction (memory) due to the development of memory B cells. These are intermediate, differentiated B cells with the capability to quickly become plasma cells. Circulating antibodies recognize antigen in tissue fluids and serum. This activity describes the physiology and pathophysiology of immunoglobulins
The advent of the polymerase chain reaction (PCR) radically transformed biological science from the time it was first discovered (Mullis, 1990). For the first time, it allowed for specific detection and production of large amounts of DNA. PCR-based strategies have propelled huge scientific endeavors such as the Human Genome Project. The technique is currently widely used by clinicians and researchers to diagnose diseases, clone and sequence genes, and carry out sophisticated quantitative and genomic studies in a rapid and very sensitive manner. One of the most important medical applications of the classical PCR method is the detection of pathogens. In addition, the PCR assay is used in forensic medicine to identify criminals. Because of its widespread use, it is important to understand the basic principles of PCR and how its use can be modified to provide for sophisticated analysis of genes and the genome
Recently, the advantages of biopolymers over conventional plastic polymers are unprecedented, provided that they are used in situations in which they raise the functionality and generate extra benefits for human life. Therefore, biopolymers have received much attention because they play an important place in day-to-day life for their specific tunable characteristics, making them attractive in a wide range of applications. Biopolymers can produce materials with tunable properties such as biodegradability, biocompatibility, renewability, inexpensiveness, availability, which are critically important for designing materials for use in biomedical applications. In addition to these properties, smart biopolymers could be prepared by changing the polymer components, which would create more target oriented applications. Biopolymers are potentially used in biomedical applications, including drug delivery, infections, tissue engineering, wound healings, and other as wells.
Quantitative estimation of protein Likhith KLIKHITHK1
Proteins are polypeptide structures consisting of one or more long chains of amino acid residues. They carry out a wide variety of organism functions, including DNA replication, transporting molecules, catalyzing metabolic reactions, and providing structural support to cells. A protein can be identified based on each level of its structure. Every protein at least contains a primary, secondary, and tertiary structure. Only some proteins have a quaternary structure as well. The primary structure is comprised of a linear chain of amino acids. The secondary structure contains regions of amino acid chains that are stabilized by hydrogen bonds from the polypeptide backbone. These hydrogen bonds create alpha-helix and beta-pleated sheets of the secondary structure. The three-dimensional shape of a protein, its tertiary structure, is determined by the interactions of side chains from the polypeptide backbone. The quaternary structure also influences the three-dimensional shape of the protein and is formed through the side-chain interactions between two or more polypeptides. Each protein at least contains a primary, secondary, and tertiary structure. Only some proteins have a quaternary structure as well.
Accurate protein quantitation is essential to protein studies in a multitude of research topics. A wide array of different methods have been developed to quantitate both complex mixtures of proteins as well as a single type of protein.
Total protein quantitation methods comprise traditional methods such as the measurement of UV absorbance at 280 nm, Bicinchoninic acid (BCA) and Bradford assays, as well as alternative methods like Lowry or novel assays developed by commercial suppliers, which often provide a well-designed, convenient kit for each type of the assay. Individual protein quantitation methods include enzyme-linked immunosorbent assay (ELISA), western blot analysis, and more recently, mass spectrometry, among others. Accurate protein quantitation is essential to all experiments related to proteins studies in a multitude of research topics. Different wide array of different methods have been developed to quantitate both complex mixtures of proteins in a given assay for total protein content and as well as for a single type of protein.
Transparent unstained samples do not absorb light and are called phase objects. When light passes through a sample area with no phase object, there is no significant change in the refractive index or optical path length. Non-diffracted light is referred to as direct or zero-order light as it continues unchanged through the sample. On the other hand, when the light passes through an area of the sample with a phase object, small changes in the refractive index will diffract and scatter some light and cause changes to the optical path length, depending on the thickness and refractive index of each structure. Thicker the structure, the greater the diffraction of the light. The diffracted light represents only a small part of the total light that has passed through the sample. This diffracted light arrives at the detector out of phase with the direct light. The small phase shift created by this, is not enough to cause great interference between the direct and diffracted light. Which along with the low absorption of transparent structures means there is negligible amplitude difference between areas where such structures are present and where they are not. Phase-contrast microscopy is a method that manipulates this property of phase objects to introduce additional interference between the direct and diffracted light. This method transforms differences in phase into differences in brightness, increasing contrast in images of non-absorbing samples.
In light microscopy, illuminating light is passed through the sample as uniformly as possible over the field of view. For thicker samples, where the objective lens does not have sufficient depth of focus, light from sample planes above and below the focal plane will also be detected. The out of focus light will add blur to the image reducing the resolution. In fluorescence microscopy, any dye molecules in the field of view will be stimulated, including those in out-of-focus planes. Confocal microscopy provides a means of rejecting the out-of-focus light from the detector such that it does not contribute blur to the images being collected. This technique allows for high-resolution imaging in thick tissues.
In a confocal microscope, the illumination and detection optics are focused on the same diffraction limited spot in the sample, which is the only spot imaged by the detector during a confocal scan. To generate a complete image, the spot must be moved over the sample and data collected point by point.
A significant advantage of the confocal microscope is the optical sectioning provided, which allows for 3D reconstruction of a sample from high-resolution stacks of images. The primary functions of a confocal microscope are to produce a point source of light and reject out-of-focus light, which provides the ability to image deep into tissues with high resolution, and optical sectioning for 3D reconstructions of imaged samples. The basic principle include illumination and detection optics are focused on the same diffraction-limited spot, which is moved over the sample to build the complete image on the detector. The entire field of view is illuminated during confocal imaging, anything outside the focal plane contributes little to the image, lessening the haze observed in standard light microscopy with thick and highly-scattering samples, and providing optical sectioning.
Atomic force microscope (AFM) is a scanning near-field tool for nanoscale investigation which was invented in 1986. Instead of using light or electron beam, AFM uses a sharp tip to ‘‘feel’’ samples. As the tip radius of curvature is on the order of nanometers, AFM can detect changes at a spatial resolution up to sub nanometer level. Compared to the optical microscope, AFM has a much higher spatial resolution which provides the ability to investigate ultrafine structure of samples and even map the distribution of single molecules.
As AFM utilizes direct contact between the tip and the sample, minimum or even no sample preparation is required.
Moreover, AFM can investigate samples in liquid which provides an opportunity to monitor samples close to their native surroundings. Further, AFM provides true 3D images. With optical and electron microscopies, only limited ranges in heights can be ‘‘in-focus’’ at any one time. Therefore, AFM can provide unique insight into the structure and functional behavior of materials. AFM is a versatile technique. Besides scanning the topography of a sample, it can also be used to investigate the mechanical properties of the sample as well as the interactions between the tip and the sample. AFM has been successfully applied in widespread branches of science and technology such as nanofabrication, material science, chemical and drug engineering, biotechnology and microbiology. As for above mentioned reasons, Atomic force microscope (AFM) is considered a useful tool for the nanoscale measurement in material-polymer science and engineering. AFM lacks the robust ability to chemically characterize materials.
Fluorescence as a phenomenon is part of a larger family of related luminescent processes in which a susceptible substance absorbs light, only to reemit light (photons) from electronically excited states after a given time.
Photo luminescent processes that are generated through excitation, whether this is via physical, mechanical, or chemical mechanisms, can generally be subdivided into fluorescence and phosphorescence. Absorption of a light quantum (blue) causes an electron to move to a higher energy orbit. After residing in this “excited state” for a particular time, the fluorescence lifetime, the electron falls back to its original orbit and the fluorochrome dissipates the excess energy by emitting a photon (green).
Compounds that display fluorescent properties are generally termed fluorescent probes or dyes. Often ‘fluorochrome’ and ‘fluorophore’ are used interchangeably. The term ‘fluorophore’ refers to fluorochromes that are conjugated covalently or through adsorption to biological macromolecules, such as nucleic acids, lipids, or proteins. Fluorochromes come in different flavors and include organic molecules (dyes), inorganic ions (e.g., lanthanide ions such as Eu, Tb, Yb, etc.)fluorescent proteins (e.g., green fluorescent protein) atoms (such as gaseous mercury in glass light tubes).
Recently, inorganic luminescent semiconducting nanoparticles, quantum dots, have been introduced as labels for biological assays, bio-imaging applications, and theragnostic purposes (the combination of diagnostic and therapeutic modalities in one and the same particle).
Fluorescence microscopy provides an efficient and unique approach to study fixed and living cells because of its versatility, specificity, and high sensitivity.
Fluorescence microscopes can both detect the fluorescence emitted from labeled molecules in biological samples as images or photometric data from which intensities and emission spectra can be deduced. By exploiting the characteristics of fluorescence, various techniques have been developed that enable the visualization and analysis of complex dynamic events in cells, organelles, and sub-organelle components within the biological specimen.
The most common techniques are
Fluorescence recovery after photo bleaching (FRAP)
Fluorescence loss in photo bleaching (FLIP)
Fluorescence localization after photo bleaching (FLAP)
Fluorescence resonance energy transfer (FRET)
Transmission electron microscope (TEM) Likhith KLIKHITHK1
Microscopy is a means by which an object is transformed in to magnified image. There are different ways for magnifying the images of very small objects by large amounts. In any type of microscopy (optical microscopy or electron microscopy), a wave of wavelength λ (light wave or electron wave) interacts with the matter and as a result of this interaction we get the
microstructural information about the object. As the study of the materials at the nano-metric level is drawing much attention of the researchers in the current era, Electron Microscopy becomes a very important physical characterization tool at the nano-metric level. Electron Microscopy stands far ahead of the optical microscopy as it can provide the much improved
resolution and depth of focus compared to optical microscopy. This is a very introductory report on the basics of the electron microscopy (particularly on Transmission electron microscopy). Transmission electron Microscopy (TEM) operates on the same basic principles as the light microscope but uses electrons as “light source” and their much lower wavelength makes it possible to get a resolution thousand times better than with a light Microscopy.
Scanning electron microscopy (SEM) Likhith KLIKHITHK1
Scanning Electron Microscope functions exactly as their optical counterparts except that they use a focused beam of electrons instead of light to “image” the specimen and gain information as to its structure and composition. Given sufficient light, the unaided human eye can distinguish two points 0.2 mm apart. If the points are closer together, they will appear as a single point. This distance is called the resolving power or resolution of the eye. Similarly, light microscopes use visible light (400- 700nm) and transparent lenses to see objects as small as about one micrometer (one millionth of a meter), such as a red blood cell (7 μm) or a human hair (100 μm). Light microscope has a magnification of about 1000x and enables the eye to resolve objects separated by 200 nm. Electron Microscopes were developed due to the limitations of light microscopes, which are limited by the physics of light. Electron Microscopes are capable of much higher magnifications and have a greater resolving power than a light microscope, allowing it to see much smaller objects at sub cellular, molecular and atomic level. The smallest the wavelength of the illuminating sources is the best resolution of the microscope. De Broglie defined the wavelength of moving particles (electron) λ = h/mv, Where λ= wavelength of particles, h= Planck, s constant, m= mass of the particle (electron), v= velocity of the particles; after substituting the known values, λ = 12.3 Ao/V. The resolution of an optical microscope is defined as the shortest distance between two points on a specimen that can still be distinguished by the observer or camera system as separate entities. Resolution (r) = λ/ (2NA), Where λ is the imaging wavelength, NA is objective numerical aperture. Magnification is the process of enlarging the appearance, not physical size, of something. Magnification is defined as the ratio of image distance versus object distance. M= v/u, Where M= magnification, u= object distance, v= image distance. Magnification is also defined as the ratio of the resolving power of the eye to resolving power (δ) of the microscope M= δ eye/ δ microscope.
Crystallography and X-ray diffraction (XRD) Likhith KLIKHITHK1
Atoms in materials are arranged into crystal structures and microstructures.
Periodic arrangement of atoms depends strongly on external factors such as temperature, pressure, and cooling rate during solidification. Solid elements and their compounds are classified into amorphous, polycrystalline, and single crystalline materials. The amorphous solid materials are isotropic in nature because their atomic arrangements are not regular and possess the same properties in all directions. In contrast, the crystalline materials are anisotropic because their atoms are arranged in regular and repeated pattern, and their properties vary with direction. The polycrystalline materials are combinations of several crystals of varying shapes and sizes. The properties of polycrystalline materials are strongly dependent on distribution of crystals sizes, shapes, and orientations within the individual crystal. Diffraction pattern or intensities of X-ray diffraction techniques are used for characterizing and probing arrangement of atoms in each unit cell, position of atoms, and atomic spacing angles because of comparative wavelength of X-ray to atomic size.The X-ray diffraction, which is a non-destructive technique, has wide range of material analysis including minerals, metals, polymers, ceramics, plastics, semiconductors, and solar cells. The technique also has wide industry application including aerospace, power generation, microelectronics, and several others. The X-ray crystallography remained a complex field of study despite wide industrial applications.
Fourier transform infrared spectroscopy (FTIR) is a largely used technique to identify the functional groups in the materials (gas, liquid, and solid) by using the beam of infrared radiations. An infrared spectroscopy measures the absorption of IR radiation made by each bond in the molecule and as a result gives spectrum which is commonly designated as % transmittance versus wavenumber (cm−1). The IR region is at lower energy and higher wavelength than the UV-visible light and has higher energy or shorter wavelength than the microwave radiations. For the determination of functional groups in a molecule, it must be IR active. An IR active molecule is the one which has dipole moment. When the IR radiation interacts with the covalent bond of the materials having an electric dipole, the molecule absorbs energy, and the bond starts back and forth oscillation. Therefore, the oscillation which cause the change in the net dipole moment of the molecule should absorb IR radiations.
A single atom doesn’t absorb IR radiation as it has no chemical bond.
Symmetrical molecules also do not absorbed IR radiation, because of zero dipole moment. For example, H2 molecule has two H atoms; both cancel the effect of each other and giving zero dipole moment to H2 molecule. Therefore, H2 molecule is not an IR active molecule. On the other hand, HF is an IR active molecule, because when IR radiation interacts with HF molecule, the charge transferred toward the fluorine atom and as a result fluorine becomes partial negative and hydrogen becomes partial positive, giving net dipole moment to H-F molecule. A particular IR radiation will be absorbed by a particular bond in the molecule, because every bond has their particular natural vibrational frequency. For example, a molecule such as acetic acid (CH3COOH) containing various bonds (C-C, C-H, C-O, O-H, and C=O), all these bonds are absorbed at specific wavelength and are not affected by other bond. In general we can say that two molecules with different structures don’t have the same infrared spectrum, although some of the frequencies might be same.
UV-visible spectroscopy is a fast analytical technique that measures the absorbance or transmittance of light. Although the UV wavelength ranges from 100–380 nm and the visible component goes up to 800 nm, most of the spectrophotometers have a working wavelength range between 200–1100 nm.
The practical range for UV-vis spectroscopy varies from 200–800 nm; above 800 nm is infrared, while below 200 nm is known as vacuum UV. The ability of matter to absorb and to emit light is what defines its color and the human eye is capable of differentiating up to 10 million unique colors. Light passes through media (transmission), reflects off both opaque and transparent surfaces, and is refracted by crystals. Covalently unsaturated compounds with electronic transition energy differences equivalent to the energy of the UV-visible light absorb at specific wavelengths. These compounds are known as chromophores and are responsible for their color. Covalently saturated groups that do not absorb UV-visible electromagnetic radiation but affect the absorption of chromophore groups are called auxochromes. When UV-vis radiation hits chromophores, electrons in the ground state jump to an excited state, which we refer to as electron-excitation, while auxochromes are electron-donating and have the capacity to affect the color of choromophores while they do not change color themselves. Water and alcohols are mostly transparent and do not absorb in the UV-vis range and so are excellent mediums for UV-visible spectroscopy. Acetone and dimethylformamide (DMF) are good solvents for compounds insoluble in water and alcohol, but they absorb light below 320 and 275 nm, respectively, so are appropriate only above these cut-off wavelengths.
Enzymes principles and applications Likhith KLIKHITHK1
Enzymes are biological catalysts (also known as biocatalysts) that speed up biochemical reactions in living organisms. They can also be extracted from cells and then used to catalyse a wide range of commercially important processes. For example, they have important roles in the production of sweetening agents and the modification of antibiotics, they are used in washing powders and various cleaning products, and they play a key role in analytical devices and assays that have clinical, forensic and environmental applications. The word ‘enzyme’ was first used by the German physiologist Wilhelm Kühne in 1878, when he was describing the ability of yeast to produce alcohol from sugars, and it is derived from the Greek words en (meaning ‘within’) and zume (meaning ‘yeast’).In the late nineteenth century and early twentieth century, significant advances were made in the extraction, characterization and commercial exploitation of many enzymes, but it was not until the 1920s that enzymes were crystallized, revealing that catalytic activity is associated with protein molecules. For the next 60 years or so it was believed that all enzymes were proteins, but in the 1980s it was found that some ribonucleic acid (RNA) molecules are also able to exert catalytic effects.These RNAs, which are called ribozymes, play an important role in gene expression. In the same decade, biochemists also developed the technology to generate antibodies that possess catalytic properties. These so-called ‘abzymes’ have significant potential both as novel industrial catalysts and in therapeutics. Notwithstanding these notable exceptions, much of classical enzymology, and the remainder of this essay, is focused on the proteins that possess catalytic activity.As catalysts, enzymes are only required in very low concentrations, and they speed up reactions without themselves being consumed during the reaction.
Cheese is a very popular food produced worldwide from the milk of ruminants using a combination of physical treatments. Key milk components for the transformation of milk into curd are casein and calcium. The majority of cheese varieties are based on the curd of modified casein micelles that result from the enzymatic rennet clotting of milk in the presence of calcium ions. The remarkable ability of the spontaneous syneresis of rennet-induced curds can be adjusted to the desired level by biological acidification, cutting, stirring, heating, pressing and salting in order to achieve the desired level of water removal in the form of whey. The mode of curdling (acid or rennet coagulation), the conditions, and the combinations of curd treatments result in numerous cheese varieties with different appearances, textures, flavors and shelf lives. Moreover, most of them are kept under specific temperature and humidity conditions to ripen for a short or a considerable amount of time. The classification of cheese varieties is not unambiguous and can be based on various criteria. For example, cheeses can be classified according to their moisture content related to yield and shelf life or according to specific features related to the treatments applied during cheesemaking and ripening. During ripening, the main solid constituents of young cheese—fat and caseins—undergo changes that increase the concentration of small size compounds in cheese—such as peptides, amino acids, small volatile molecules—control moisture loss and configure textural properties. In particular, the compounds of mature cheese flavor result from complicated biochemical pathways that take place during cheese ripening or even storage.
The function of the fermenter or bioreactor is to provide a suitable environment in which an organism can efficiently produce a target product—the target product might be cell biomass,metabolite and bioconversion Product. It must be so designed that it is able to provide the optimum environments or conditions that will allow supporting the growth of the microorganisms. The design and mode of operation of a fermenter mainly depends on the production organism, the optimal operating condition required for target product formation, product value and scale of production.
The choice of microorganisms is diverse to be used in the fermentation studies. Bacteria, Unicellular fungi, Virus, Algal cells have all been cultivated in fermenters. Now more and more attempts are tried to cultivate single plant and animal cells in fermenters. It is very important for us to know the physical and physiological characteristics of the type of cells which we use in the fermentation. Before designing the vessel, the fermentation vessel must fulfill certain requirements that is needed that will ensure the fermentation process will occur efficiently. Some of the actuated parameters are: the agitation speed, the aeration rate, the heating intensity or cooling rate, and the nutrients feeding rate, acid or base valve. Precise environmental control is of considerable interest in fermentations since oscillations may lower the system efficiency, increase the plasmid instability and produce undesirable end products.
Penicillin is one of the most commonly used antibiotics globally, as it has a wide range of clinical indications. Penicillin is effective against many different types of infections involving gram-positive cocci, gram-positive rods (e.g., Listeria), most anaerobes, and gram-negative cocci (e.g., Neisseria). Importantly, certain bacterial species have obtained penicillin resistance, including enterococci. Enterococci infections now receive treatment with a combination of penicillin and streptomycin or gentamicin. Certain gram-negative rods are also resistant to penicillin due to penicillin’s poor ability to penetrate the porin channel. However, later generations of broad-spectrum penicillins are effective against gram-negative rods. Second-generation penicillins (ampicillin and amoxicillin) can also penetrate the porin channel, making these drugs effective against Proteus mirabilis, Shigella, H. influenzae, Salmonella, and E. coli. Third-generation penicillins such as carbenicillin and ticarcillin are also able to penetrate gram-negative bacterial porin channels. Fourth-generation penicillins such as piperacillin are effective against the same bacterial strains as third-generation penicillins as well as Klebsiella, enterococci, Pseudomonas aeruginosa, and Bacteroides fragilis.
Beer is one of the oldest and most widely consumed alcoholic drinks in the world, and the third most popular drink overall after water and tea. Beer is brewed from cereal grains most commonly from malted barley, though wheat, maize (corn), and rice are also used. The process of beer production is known as brewing. Word brewing is derived from “Bieber” its means to drink.
Brewing is a complex fermentation process. It differs from other industrial fermentation because flavor, aroma, clarity, color, foam production, foam stability and percentage of alcohol are the factors associated with finished product.
During the brewing process, fermentation of the starch sugars in the wort produces ethanol and carbonation in the resulting beer. Most modern beer is brewed with hops, which add bitterness and other flavors and act as a natural preservative and stabilizing agent. Other flavoring agents such as gruit, herbs, or fruits may be included or used instead of hops.
Chromatography is based on the principle where molecules in mixture applied onto the surface or into the solid, and fluid stationary phase (stable phase) is separating from each other while moving with the aid of a mobile phase.
The factors effective on this separation process include molecular characteristics related to adsorption (liquid-solid), partition (liquid-solid), and affinity or differences among their molecular weights
Because of these differences, some components of the mixture stay longer in the stationary phase, and they move slowly in the chromatography system, while others pass rapidly into mobile phase, and leave the system faster.
The word "vitamin" comes from the Latin word “vita”, means "life". Vitamins are organic components in food that are required in very small amounts for growth and for maintaining good health. Vitamins are chemicals found in very small amounts in many different foods Vitamins and minerals are measured in a variety of ways. The most common are:
mg – milligram (a milligram is one thousandth of a gram)
mcg – microgram (a microgram is one millionth of a gram. 1,000 micrograms is equal to one milligram)
IU – international unit (the conversion of milligrams and micrograms into IU depends on the type of vitamin or drug)
(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.
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.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
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.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
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.
2. Contents
Introduction
Definition and classification of carbohydrates
Monosaccharides
Disaccharides
Oligosaccharides
Polysaccharides
Quantitative analysis of carbohydrates
Anthrone method
Nelson Somogyi method
Dinitrosalicylic acid method (DNSA)
Conclusion
Reference
3. INTRODUCTION
Carbohydrates, which are made up of carbon, hydrogen, and oxygen, are organic compounds that serve as a
source of energy for microorganism, plants and animals.
The main monosaccharide is glucose (fig 1), which is utilized as an energy source by most living organisms.
Glucose can be derived from starch and sugars in the diet, from glycogen that is stored in the body, or synthesized
from the carbon skeleton of amino acids, lactate, glycerol, or propionate via gluconeogenesis (fig 2).
The brain preferentially uses glucose as its main source of energy, and glucose is the required energy source for
red blood cells and other cells with few or no mitochondria.
Figure 1
5. The fate of ingested carbohydrates in an animal is determined by the monomeric composition of the
carbohydrate, the types of linkages among monomers, and the degree of polymerization.
Digestible carbohydrates include monosaccharides, disaccharides, starch, and glycogen.
Only monosaccharides can be absorbed from the small intestine, but glycosidic linkages in
disaccharides, starch, and glycogen may be hydrolyzed by endogenous enzymes in the small intestine,
resulting in release of their constituent monosaccharides.
However, these enzymes show high specificity to their target sugar units (substrate), which
consequently results in only a limited number of carbohydrates in the feed that can be digested by the
animal.
Non-digestible carbohydrates that reach the large intestine may be digested by microbial enzymes
because intestinal microorganisms secrete glycoside hydrolases and polysaccharide lyases that humans
and other mammals do not express.
Non-digestible carbohydrates include oligosaccharides, resistant starch, and non-starch
polysaccharides and are collectively known as fiber.
The large differences in the physical properties of carbohydrates make it difficult to analyze fiber and
non-digestible carbohydrates.
6. Dietary fiber may be divided according to solubility.
Soluble dietary fiber (SDF) may be partially or completely fermented by the microbiota in the large intestine,
producing short-chain fatty acids (SCFA), which include acetate, propionate, and butyrate.
Insoluble dietary fiber (IDF) may also be fermented, but to a lesser extent than SDF.
Fermentation of dietary fiber is a major source of energy in ruminants and hindgut fermenters, but only to a lesser
extent in pigs and poultry.
The relationship between the host and the gut microbiota is symbiotic.
As microorganisms ferment non-digestible carbohydrates, endogenous mucosal secretions, and exfoliated
epithelial cells to utilize the carbon and N to sustain themselves, SCFA and lactate are produced and absorbed by
the animal.
Estimation of carbohydrates is very important in laboratory practices to quantify the amount of sugar in any give
biological sample.
It gives out preliminary data about the sugar composition in given sample
Carbohydrate estimation have a great role in medical, agriculture and food industry.
7. DEFINITION AND CLASSIFICATION OF CARBOHYDRATES
Classification(fig 3) based on molecular size or Degree of Polymerization, carbohydrates are grouped into
1. Monosaccharides
2. Disaccharides
3. Oligosaccharides
4. Polysaccharides
Figure 3
8. Monosaccharides are chiral, poly-hydroxylated aldoses or ketoses that cannot be hydrolyzed into smaller
carbohydrate units.
They can be classified according to the number of carbon atoms in their structure (fig 4), which range from three
to nine carbon atoms (i.e., triose, tetrose, pentose, hexose, heptose, octose, and nonose), by the type of carbonyl
group they contain (i.e., aldose or ketose), and by their stereochemistry (i.e., D or L (fig 1)), and they have the
general chemical formula (CH2O)n.
Figure 4
9. Aldoses are referred to as reducing sugars because of their reducing effect on certain ions or compounds,
oxidizing their aldehyde group to a carbonyl group.(fig 5)
The simplest aldose sugar with a chiral atoms glyceraldehyde, with its second C molecule attached to four
different groups, giving the ability for this C to have two spatial configurations, and glyceraldehyde therefore
exist in both the D- and the L- forms.(fig 6)
Chiral carbon atoms have each of their four tetrahedral bonds connected to a different group.
The chirality of sugars and amino acid are commonly designated by the D/L system and is named in relation to
the structure of glyceraldehyde.
Figure 5
Figure 6
10. 1. Monosaccharides
The most common monosaccharides are the 6-C aldohexoses, which include the aldohexose D-glucose, and are
usually present in their ring structures called a pyranose ring rather than in open-chain structures (fig7).
Figure 7
11. In oligo- and polysaccharides, aldo-pentoses can occur as a 5-C ring structure known as a furanose ring(fig 8).
D-Glucose, considering all of its combined forms, is the most abundant monosaccharide that naturally occurs in
nature.
The most abundant ketose is D-arabino-hexulose, known more commonly by its trivial name, D-fructose. (fig 9)
The three trioses include ketose dihydroxyacetone and both enantiomeric forms of glyceraldehyde.
Erythrose and threose are examples of tetroses, and pentoses include ribose, arabinose, xylose, and apiose.
Figure 8 Figure 9
12. Sugars, such as glucose, galactose, mannose, and fructose, which have different structures, but have the same
chemical formula, C6H12O6, are called isomers. (fig 10)
Sugars that differ in configuration around only one carbon atom are called epimers, such as D-glucose and D-
mannose, which vary in their structures around C-2(fig 11).
Figure 10 Figure 11
13. A pair of enantiomers is a special type of isomerism where the two members of the pair are mirror images of
each other and are designated as being in the D- or L- structure (i.e., D-glucose or ʟ-glucose(fig 1)), depending
on the position of the –OH group linked to the asymmetric carbon farthest from the carbonyl group.
Other types of monosaccharides include alditols, or polyols, which are aldoses or ketoses that had their carbonyl
groups reduced to an alcohol.
An example of a naturally occurring alditol in plants and other organisms is D-glucitol, known commonly as
sorbitol(fig12), which is the product of the reduction of D-glucose.
Figure 12
14. Absorption and metabolism of polyols vary among types, but most are fermented in the large intestine.
Deoxy sugars are missing one or more hydroxyl groups attached to their carbon atoms, such as 6-deoxy-L-
mannose (L-rhamnose), which is commonly associated with pectin, 2-deoxy- D-ribose, the sugar component of
DNA, and 6-deoxy-L-galactose (L-fucose), a component of glycoproteins and glycolipids in cell walls and
mammalian cells. (Fig13)
Figure 13
15. Uronic acids are sugar acids in which the terminal –CH2OH group undergoes oxidation to yield a carboxylic
acid.(fig 14)
Uronic acids that contribute to dietary fiber include constituents of non-digestible polysaccharides of plants and
algae, such as D-glucuronic acid, D-galacturonic acid and D-mannuronic acid. (fig 15)
Sugar from the activated form of glucuronic acid is used in the synthesis of glycosaminoglycans in mammals, and
L-iduronic acid is synthesized from D-glucuronic acid after it has been incorporated into the carbohydrate chain.
Figure 14 Figure 15
16. 2. Disaccharides
Two monosaccharide units joined by an acetal or ketal linkage is referred to as a disaccharide.
A glycosidic bond joins 2 monosaccharide units and it can either be an α-glycosidic bond (A) if the anomeric
hydroxyl group of the sugar is in the α configuration or a β-glycosidic bond (B) if it is in the β configuration. (fig
16)
A glycosidic bond is named according to the position of the carbon atom being linked, for example, an α-
glycosidic bond connecting C-1 of a glucose molecule and C-4 of another glucose molecule in maltose is called
an α-(1,4) glycosidic bond.
Figure 16
A B
17. The three most common disaccharides (fig 17)are
1. Maltose
2. Lactose
3. Sucrose
Maltose is a reducing sugar that is a product of the hydrolysis of starch by the enzyme α-amylase.
Lactose is a reducing sugar that consists of a D-glucosyl unit and an α-D-galactopyranosyl unit linked by a β-
(1,4) glycosidic bond and is present in milk and milk products such as skim milk and whey.
Sucrose is made up of a glucose and a fructose linked by an α-(1,2) glycosidic bond.
Contrary to the general head-to-tail linkage (anomeric carbon atom to carbon atom containing a hydroxyl group)
in the structure of oligo and polysaccharides, in sucrose the glycosidic bond linking an α-D-glucopyranosyl unit
and a β-D-fructofuranosyl unit is in a head-to-head fashion (anomeric carbon atom to anomeric carbon atom)
making it a non-reducing sugar.
Sucrose is synthesized through the process of photosynthesis to provide energy and carbon atoms for the
synthesis of other compounds in the plant.
Figure 17
18. Maltose, lactose, and sucrose are hydrolyzed into their constituent monosaccharide units by the enzymes maltase,
lactase, and invertase, respectively.(fig 18)
The α-glucosidases maltase-glucoamylase and sucrase-isomaltase complexes that are present in the
brush border of the small intestine cleave the glycosidic bonds in maltose and sucrose, respectively,
with most of the maltase activity coming from the sucrase-isomaltase complex.
The monosaccharides that result from the digestion of these disaccharides are readily absorbed in the
small intestine.
Lactase, a β-galactosidase, also is expressed by young mammals that digest lactose into its constituent
monosaccharides that are subsequently absorbed in the small intestine.
Figure 18
19. Other disaccharides that are present in nature include trehalose, cellobiose, and gentiobiose.(fig 19)
Trehalose is a nonreducing disaccharide made up of two α-D-glucopyranosyl units linked together by
an α-(1,1) (A) glycosidic bond.
Trehalose is found in small amounts in mushrooms, yeasts, honey, certain seaweeds, and invertebrates
such as insects, shrimps, and lobsters.
Trehalose is digested by the α-glucosidase enzyme trehalase, which is expressed in the small intestine
of humans and most animals.
Two glucose molecules are linked together by a β-(1,4) (B)and a β-(1,6) (C) glycosidic bonds to form
cellobiose and gentiobiose, respectively, and these disaccharides can be utilized only after microbial
fermentation because mammals lack the enzymes capable of digesting these bonds.
Figure 19
A B C
20. 3. Oligosaccharides
Oligosaccharides consist (fig 20) of
1. Galacto-oligosaccharides
2. Fructo-oligosaccharides
3. Mannan-oligosaccharides
that cannot be digested by pancreatic or intestinal enzymes, but are soluble in 80% ethanol.
B
A
C
Figure 20 (A,B,C)
21. Galacto-oligosaccharides, or α-galactosides, that are present in large amounts in legumes, are comprised of
raffinose, stachyose, and verbascose, which have a structure consisting of a unit of sucrose linked to one, two, or
three units of D-galactose, respectively.
These oligosaccharides cause flatulence in humans due to the lack of an enzyme, α-galactosidase, that
hydrolyzes the glycosidic bonds linking the monosaccharides that constitute these α-galactosides and are,
therefore, utilized by bacteria in the large intestine.
In raffinose, (fig 21) D-galactose is linked to sucrose by an α-(1,6) bond, whereas two units and three units of D-
galactose are linked to sucrose, also via α-(1,6) glycosidic bonds, in stachyose and verbascose, respectively.
Figure 21
22. Transgalacto-oligosaccharides are another type of galacto-oligosaccharides that may have prebiotic effects in
humans and are commercially synthesized from the transglycosylation actions of β-glycosidases on lactose,
creating β-(1,6) polymers of galactose linked to a terminal glucose unit via an α-(1,4) glycosidic bond.
However, transgalacto-oligosaccharides are not naturally synthesized.
Fructo-oligosaccharides, or fructans, are chains of fructose monosaccharides with a terminal glucose unit and are
classified as inulins or levans. (fig 22)
Figure 22
23. Inulin is mostly found in dicotyledons, whereas levans are mainly found in monocotyledons.
Fructo-oligosaccharides are not hydrolyzed in the small intestine due to the β-linkages between their monomers,
but can be fermented to lactic acid and SCFA in the large intestine.
Inulin occurs naturally in onions, garlic, asparagus, bananas, wheat, and chicory as a storage carbohydrate.
Inulin is made up of β-D-fructofuranosyl units linked by β-(2,1) glycosidic linkages and have a Degree of
Polymerization that ranges from 2 to 60.
The polymer is composed of fructose residues present in the furanose ring form and often have a terminal
sucrose unit at the reducing end.
Levans are fructans that have an average length of 10 to 12 fructose units linked by β-(2,6) linkages, but can
have a Degree of Polymerization of more than 100,000 fructose units and are found in bacterial fructans and in
many monocotyledons.
Levans are derived from the transglycosylation reactions catalyzed by the enzyme levansucrase that is secreted
by certain bacteria and fungi that preferentially use the D-glycosyl unit of sucrose, thereby converting sucrose to
levans with β-(2,1) linked side-chains.
Polysaccharides containing a significant number of β-(2,1) linkages also can be referred to as “levan”.
24. A third type of fructans, called graminantype fructans, contain a combination of both β-(2,1) and β-(2,6) linkages
and are present in wheat and barley.
Mannan-oligosaccharides are composed of polymers of mannose that are derived from yeast cell walls, and are
located on the outer surface of yeast cell walls attached to β-glucans of the inner matrix via β-(1,6) and β-(1,3)
glycosidic linkages.
Mannan-oligosaccharides and fructo-oligosaccharides may behave as prebiotics due to their beneficial health
effects on the host by stimulating the growth or activity of certain bacteria in the large intestine.
4. Polysaccharides
Polysaccharides are high-molecular-weight carbohydrates that are polymers of monosaccharides.
Polysaccharides are made up of sugar polymers that vary in size and may either be linear or branched.
The Degree of Polymerization varies with the type of polysaccharide and may range from 7,000 to 15,000 in
cellulose and up to more than 90,000 in amylopectin.
Polysaccharides can be classified as homo-polysaccharides if they contain only one type of sugar residue (e.g.,
starch, glycogen, and cellulose) or as hetero-polysaccharides if they contain two or more different kinds of sugar
residues in their structure (e.g., arabinoxylans, glucomannans, and hyaluronic acid).(fig 23)
In humans polysaccharides are present in large quantities, and are divided into starch and glycogen and non-
starch polysaccharides.
25. Starch can be linear or branched and is the storage form of carbohydrates in plants, whereas glycogen is highly
branched and is present only in animal tissue, primarily in the muscle and liver.
Starch is one of the most abundant carbohydrates in nature.
It is synthesized to store energy for plant growth and is stored in seeds, tubers, roots, stems, leaves, and some
fruits.
Starch is a polymer of D-glucose that is comprised of two types of molecules, amylose and amylopectin (fig 24).
Figure 23
26. Figure 24
Amylose is a short linear polymer of glucose with an average Degree of Polymerization of 1,000 glucose units
linked via α-(1,4) bonds.
Amylopectin contains larger chains of glucose with Degree of Polymerization of 10,000 to 100,000 with branch
points at the α-(1,6) linkages for every 20 to 25 glucose units.
The total number of α-(1,6) linkages are only about 4 to 5 % of the total glycosidic bonds in amylopectin.
Native starch contains both forms as semi-crystalline granules of varying proportions of amylose and
amylopectin, depending on the plant source.
Starch granules have varying structural and chemical compositions depending on the plant species and the part
of the plant where it is located.
The size of the starch granules influences the surface-to-volume ratio, and the smaller the granule, the larger the
surface-to-volume ratio resulting in more surface area for enzyme hydrolysis in the digestive tract.
27. Digestion of starch begins in the mouth where salivary α-amylase is secreted, which acts only on the α-(1,4)
linked linear chains of amylose and amylopectin, until this enzyme is deactivated by the low pH in the stomach.
Large quantities of pancreatic α-amylase specific only to α-(1,4) linkages are secreted into the duodenal lumen,
producing maltose and maltotriose as the products of luminal amylose and amylopectin digestion, along with the
branched oligosaccharide α-dextrin resulting from the partial hydrolysis of amylopectin due to the inability of α-
amylase to cleave α-(1,6) linkages.
Starch digestion is completed by oligosaccharidosis (i.e., α-glucosidases) expressed by glands in the small
intestine.
These α-glucosidases include sucrose-isomaltase and maltase-glucoamylase complexes.
Both complexes have differences in their degree of specificity for the products of α-amylase digestion and
cleave the α-(1,4) and α-(1,6) bonds in α-dextrins in a complementary manner, producing free glucose that is
transported into the enterocytes.
28. Glycogen, (fig 25) an α-(1,4)-D-glucan with α-(1,6) linked branches, has a higher degree of branching compared
with amylopectin and is present in animal tissues, mainly in skeletal muscle and the liver.
The branch points of glycogen occur after an average of 8 to 10 glycosyl units.
A polymer of glycogen may contain up to 100,000 units of glucose.
Digestion of glycogen is similar to that of amylopectin, which results in glucose absorption in the small intestine.
The extensive branching of glycogen enhances its solubility, which allows glucose to be mobilized more readily.
Figure 25
29. QUANTITATIVE ANALYSIS OF CARBOHYDRATES
Carbohydrates can be analyzed by both qualitative and quantitative methods.
In case of the qualitative analysis, the presence or absence of a carbohydrate in the given sample is determined,
whereas in the quantitative analysis, the concentration (mg/ml) of the carbohydrate in the given sample is
determined or compared with a suitable reference standard compound.
The most common mode of quantitative analysis are;
1. Anthrone Method
2. Nelson-Somogyi Method
3. Dinitrosalicylic Acid (DNSA) Method
30. 1. Anthrone method
Aim: To estimate the concentration of total carbohydrates in a given sample by Anthrone method.
Principle:
Carbohydrates are first hydrolysed into simple sugars using sulphuric acid.
In hot acidic medium, glucose is dehydrated to hydroxymethyl furfural.
Anthrone reagent is used as a coloring agent that reacts with furfural derivative to form a blue-green complex.
Absorbance of these compounds is measured by a spectrophotometer at 630 nm.
31. Reagents:
Test sample
2.5 N HCl: Add 20.833mL of HCl in 100 mL of distilled water. (Prepare the solution fresh)
Anthrone reagent: Dissolve 200 mg Anthrone in 100 ml of ice cold 95% H2SO4. Prepare fresh before use.
Standard glucose Stock: Dissolve 100 mg of glucose in 100 ml water.
Working standard: 10 ml of stock diluted to 100 ml with distilled water.
Store at 4°C after adding a few drops of toluene
Procedure
Take clean and dry test tubes and mark all the tubes as per the protocol.
Pipette out 0.2-1.0 ml of glucose working standard solution in duplicate test tubes.
Make up the volume to 1 ml in each test tube by adding distilled water.
In one test tube take only 1 ml of distilled water and mark it as blank.
Then add 3 ml of Anthrone reagent to each test tube and mix thoroughly.
Heat the test tubes for 8 minutes in a boiling water bath.
Cool rapidly and read the green to dark green color at 630 nm.
Draw a standard graph by plotting concentration of the standard on the X-axis versus absorbance on the Y-axis.
From the standard graph calculate the amount of carbohydrate present in the sample
33. 2. Nelson-Somogyi Method
Aim: To estimate the concentration of reducing sugar in the given sample by Nelson-Somogyi method
Principle:
The reducing sugars when heated with alkaline copper tartrate reduce the copper from the cupric to cuprous state
and thus cuprous oxide is formed. (Reddish brown ppt)
When cuprous oxide is treated with arsenomolybdic acid, the reduction of molybdic acid to molybdenum blue
takes place.
The blue color developed is compared with a set of standards in a colorimeter at 620nm.
34. Reagents:
Alkaline Copper Tartrate
(a) Dissolve 2.5 g anhydrous sodium carbonate, 2 g sodium bicarbonate, 2.5 g potassium sodium tartrate and 20 g
anhydrous sodium sulphate in 80 mL water and make up to 100 mL.(A)
(b) Dissolve 15 g copper sulphate in a small volume of distilled water. Add one drop of sulphuric acid and make up
to 100 mL.(B)
Mix 4 mL of B and 96 mL of solution A before use.
35. Arsenomolybdate reagent:
(a)Dissolve 2.5 g ammonium molybdate in 45 mL water. Add 2.5 mL sulphuric acid and mix well. Then add 0.3
g disodium hydrogen arsenate dissolved in 25 mL water. Mix well and incubate at 37°C for 24–48 hours.
Standard glucose solution: Stock: 100 mg in 100 mL distilled water.
Working standard: 10 mL of stock diluted to 100 mL with distilled water [100 µg/mL].
Procedure:
Take clean and dry test tubes and mark all the tubes as per the protocol.
Pipette out 0.2-1.0 ml of glucose working standard solution in duplicate test tubes.
Make up the volume to 1 ml in each test tube by adding distilled water.
In one test tube take only 1 ml of distilled water and mark it as blank.
Add 1 mL of alkaline copper tartrate reagent to each tube.
Place the tubes in a boiling water for 10 minutes.
Cool the tubes and add 1 mL of arsenomolybolic acid reagent to all the tubes.
36. Make up the volume in each tube to 10 mL with distilled water.
Read the absorbance of blue color at 620 nm.
Draw a standard graph by plotting concentration of the standard on the X-axis versus absorbance on the Y-axis.
From the standard graph calculate the amount of carbohydrate present in the sample
Standard graph for Nelson-
Somogyi Method at 620nm
37. 3. Dinitrosalicylic Acid (DNSA) Method
Aim: To estimate the concentration of reducing sugar in a given sample by dinitrosalicylic acid (DNSA) method.
Principle:
For estimation of reducing sugar, dinitrosalicylic acid method is an alternative to Nelson-Somogyi method.
This method is simple, sensitive and adoptable for handling a large number of samples at a time.
This method tests for the presence of free carbonyl group (C=O), present in the so-called reducing sugars.
This involves the conversion of reducing sugar to furfural under alkaline conditions, which reduces one of the
nitro group (–NO2) of DNSA to amino group (–NH2) to produce orange brown color 3-amino-5-nitrosalicylic
acid, with absorbance maxima at 540 nm.
38. Reagents:
Test sample
Standard glucose solution: Dissolve 100 mg of glucose and make the final volume up to 100 ml with distilled water.
Dinitrosalicylic acid reagent (DNSA Reagent): Dissolve by stirring 1 g dinitrosalicylic acid, 200 mg crystalline phenol
and 50 mg sodium sulphite in 100 ml 1% NaOH.
Store at 4°C. Since the reagent deteriorates due to sodium sulphite, long storage is required, sodium sulphite may be
added at the time of use.
40% Rochelle salt solution (potassium sodium tartrate): 40g of potassium sodium tartrate in 100 of distilled water.
Procedure:
Take clean and dry test tubes and mark all the tubes as per the protocol.
Pipette out 0.2-1.0 ml of working standard solution in duplicate test tubes.
Make up the volume to 1 ml in each test tube by adding distilled water.
In one test tube take only 1 ml of distilled water and mark it as blank.
Add 3 ml of DNSA reagent.
Heat the contents in a boiling water bath for 5 min.
Add 1 ml of 40% Rochelle salt solution when the contents of the tubes are warm.
Cool and read the intensity of dark red color at 540 nm.
39. Draw a standard graph by plotting concentration of the standard on the X-axis versus absorbance on the Y-axis.
From the standard graph calculate the amount of carbohydrate present in the sample
Standard graph for
Dinitrosalicylic acid method at
540nm
40. CONCLUSION
Carbohydrates are subdivided into several categories on the basis of the number of sugar units and
how the sugar units are chemically bonded to each other.
Categories include monosaccharides, disaccharides, oligosaccharides and polysaccharides
Sugars are intrinsic in plant products and milk products.
Sugars also are added to foods during processing and preparation or at the table.
These “added sugars” (or extrinsic sugars) sweeten the flavor of foods and beverages and improve
their palatability.
Sugars are also used in food preservation and for functional properties such as viscosity, texture, body,
and browning capacity.
They provide calories but insignificant amounts of vitamins, minerals, or other essential nutrients.
The Nutrition Facts label provides information on total sugars per serving but does not currently
distinguish between sugars naturally present in foods and added sugars.
So quantification of carbohydrate are very important in food and drug industry.
All the three above mention methods of quantification are major and most common in daily practices.
Quantification give an confirm idea about the amount of carbohydrate content in given test sample.
41. REFERENCE
L. Navarro ETAL.,(2019) A review: Structures and characteristics of carbohydrates in diets fed to pigs Navarro
et al. Journal of Animal Science and Biotechnology.10:39 https://doi.org/10.1186/s40104-019-0345-6
Joanne Slavin and Justin Carlson Carbohydrates Adv Nutr. 2014 Nov; 5(6): 760–
761. doi: 10.3945/an.114.006163
Somogyi, M. (1952). J. Biol. Chem., 200, 245.
Krishnaveni, S.; Theymoli Balasubramanian and Sadasivam, S. (1984). Food Chem., 15, 229.