Polarimetry Polarimetry is used to analyze chiral substances and determine their eee aton in solutions. It is applied in quality control, laboratory analytics, as well as in R&D in the pharmaceutical, cosmetics, chemical, food, and medical industries. Polarimetry is a superior. sensitive and nondestructive measuring technique for: the measurement of optical activity. as exhibited by inorganic as well as organic compounds. The concentration énd physical properties of the solution influence the plane of polarized light and this is detected as the angle of optical rotation by a polarimeter. From_ this ‘measurement, different parameters can be defined, such as specific rotation, concentration, sugar content, and purity. Polarimetry is the key to understandin molecules are characterized by their they are called op rotation. ¢ chiral molecules in terms of optical activity. Chiral Property to rotate the plane of polarized light. Therefore, ‘ically active. The effect caused by this property is referred to as optical The geometrical property of a molecule that is anal. yzed with a polarimeter is called chirality; hence molecules with mitror-image geometries are called chiral, The cannot be Superimposed on their mirror image. The mirror images are called left and right-handed enantiomers. Chiral molecules are typically organic molecules and biomolecules, such as Sugars. starch, flavors, and essential.oils, active ph warmaceutical ingredients (APIs), amino acids. and various other biomolecules. Chiral molecules rotate the plane of Polarized light. This property is called optical Left- and right-handed enantiomers activity. ‘urn the polarization plane in opposite directions. The angle by which the polarization plane is rotated is called optical rotation. It is measured in degrees [°OR] with a polarimeter image constitution of their atoms. The tnitror image of chiral compounds cannot be superimposed with the original (see Fig, 1). This is in contrast to an achival object, as for exampl © @ symmetric flask, whose mirror image ean be superimposed with the original!!l,Principles of Polarimetry :A polarimeter consists of a polarized li ight source, an analyzer, a graduated circle to measure the rotation angle, and sample tubes. The polari through the sample tube and exhibits angular rotation to the left (-) or tight ed light passes Polarimetry is used to analyze chiral substances and solutions. It is applied in quality control, laboratory pharmaceutical, cosmetics, chemical, food, and medical determine their concentration in analytics, as well as in R&D in the industries, Chemists use polarimeters to investi gate the influence of compounds (in the sample cell) on plane polarized light. Polarimetry ty of ways. Their primary use is to measure the angle of rotation of an optically active substance using polarized light. The polarized light will either rotate clockwis ‘Se or counter-clockwise and the amount it rotates A polarimeter is defined as a rotation caused by the passi this sodium light is used b high. scientific instrument th ing of polarized light thro ecause it produces monocl at is used for measuring the angle of ugh an optically active substance. For hromatie light and the energy output is Values for specific rotatio: n are reported in units of de; shortened to just degrees, gmL-g-1-dm-1, which are typically wherein the other component 's of the unit are tacitly assumed, Components : Polarimeters have two Nicol prisms, a type of polarizer. This polarizer, a fixed prism, is used to produce a polarized beam of light. The analyzer is used to obeaun the Polarized light that is produced, and canbe rotated. The polarimeter uses 2 light source, usually a mercury or sodium discharge tube. The measuring range of MCP polarimeters extends from -89.9 °OR to +89.9 °OR. This is the range that can be unambiguously measured with a polarimeter. Yellow light (low pressure sodium bulb) is used in polarimetry because: It is a cheap and convenient light source. It is sufficiently monochromatic so that there should be one measurable optical rotation rather than multiple angles of rotation form multiple wavelengths of light. if Summary A simple, inexpensive, and flexible polarimeter was made from two LEDs, a polarizing film, a few wires, a single resistor, a battery, a few Lego bricks, and one multimeter. This polarimeter is used to demonstrate basic Properties of polarimetry such optical rotation as a function of concentration, path length, ee oo oes well as to investigate some applications such as determining t he a re ; es 7 = solution of arabinose. the amount of limonene in citrus ski wax remover, and the optical rotations of syrups, honeys, and essential oils. as nd wavelength, asefractometry is the analytical method of me: ring, substances’ refractive index (one of ir fundamental physical properties) in order to, for example, assess their composition or purity. Arefractometer is the instrument used to measure refractive index ("RI"). Although refractometers are best known for measuring liquids, they are also used to measure gases and solids; such as glass and gemstones. The RI of a substance is strongly influenced by temperature and the wavelength of light used to measure it, therefore, care must be taken to control or compensate for temperature differences and wav:length. RI measurements are usually reported at a reference temperature of 20 degrees Celsius, which is equal to 68 degrees Fahrenheit, and considered to be room temperature. A reference wavelength of 589.3 nm (the sodium D line) is most often used. Though RI is a dimensionless quantity, it is typically reported as nD20 (or n20 D), where the "n" represents refractive index, the "D" denotes the wavelength, and the 20 denotes the reference temperature. Therefore, the refractive index of water at 20 degrees Celsius, taken at tre Sodium D Line, would be reported as 1.3330 nD20. Refractometers are frequently used by grape growers and kiwifruit growers for Brix testing of sucrose levels in their fruit, Refractometry is also used in the gelatin industry. To convert the RI of a gelatin sol (reported in Brix) to a gelatin concentration, one need only multiply by eight-tenths (0.8). A sol with a 10.0 RI would therefore be 8% gelatin by weight. This is known to be a reliable conversion for gelatin sols as low as 1% up to over 50%. Types of refractometers There are four main types of fluid refractometers: traditional handheld refractometers, digital handheld refractometers, Abbe tefractometers, and inline process refractometers. Refactometer applications Refractometers are used for quality assurance in the pharmaceutical, chemical, food and beverage industries as well as in the production of flavours, perfumes and essential oils or in the petrochemical, ' metalwork and automotive industries. Generally speaking, the determination of the refractive index helps to examine the purity of a substance quickly and reliably. In addition, the progress of reactions and substance conversions can be read via the refractive index. What is a Refractometer? i g concentrations of aqueous A refractometer is a simple instrument used for measuring concentrations of qc solutions such as gases, liquids, and translucent solids. pine Peele Celera available depending on the application. Refractometers ean be handheld, compact, Pench Abbe, and Brix as well as different types for measuring materials such as salt, battery acid. Refractometers are used in the laboratory and in many indus agriculture ndustries. . food, chemical, and manufacturingIn the lab, refractometers measure the composition of samples from many industries, Some refractometers are designed for challenging samples and provide highly precise measurements. In the food industry, refractometers have many uses including measuring sugar and salt. In winemaking, Brix refractometers can check the sugar content of the grape juice. Sugar can be measured in a jelly and jam, milk, fruit juice, and other types of beverages. Salt ‘be measured in brines, cheese, or even in bakery items. content can In agricultur beekeepers use refractometers to check the moisture in honey. Refractometers can also measure the water content of milk or for determining concentration of soy milk, Refractometers can also measure chemical concentrations, validate mixtures and determine the purity of materials, in manufacturing, inline process reftactometers provide composition of substances, control dilution, and check for the purity of a substance. What does a refractometer measure? Using these values, you ean determine the concentrations of sol have diferent refractive indexes depending on their concentration in water, lutions. For example, solutions The prism in the refractometer has a greater | refractive index than the solution, Measurements are read at the point where the prism and solution meet, With a low Concentration solution, the refractive index of the prism is much greater than that of the sample, creati : low reading ("A" on diagram). The reverse would hap 'B" on diagram). What are the types of refractometers? an Handheld refractometers are the mi analog instruments that fit neatly in the palm of y operation with reliable measurements, lost commonly used. They are compact ‘our hand and provide quick and effortlessad Digital refractometers feature an easy-to-read LCD, eliminating reading errors that can occur when using analog handheld refractometers. Abbe refractometers are the larger bench laboratory refractometers that provide highly accurate measurements of refractive index and Brix measurements of liquids and solids. What is a Brix Scale and Brix percentages? ‘The Brix scale is calibrated to the number of grams of cane sugar contained in 100 mL of water. Therefore, the Brix % reading equals actual sugar concentration, DENSITOMETER Densitometry determines the optical density of a sample in solution, in-gel, or at a stage following transfer to membrane. The technique uses standalone densitometers, imaging systems, or separate software. In Wester blotting, densitometry quantitates proteins within the linear dynamic range of a chosen detection method. Detection methods include colorimetry and immunoblotting (via chemiluminescence, fluorescence, and radiolabeling). Software algorithms determine the density of signal across a selected area. This area might be a specific protein band. The software can compare the measured density to a background area, typically adjacent to the target band, to produce a relative measurement. What factors influence the reliability of densitometry? The importance of quantitation will vary depending on the specific Western blot experiment, ‘An appropriate detection method affords the level of quantitation required by the experiment. In selecting a detection method, considerations include: + Sensitivi ity: Influences the lower limit of detection for low abundance proteins. Linear dynamic range: The signal intensity range that is equivalent to protein concentration, + Signal stal * Signal-to-ncise ra Phe ability to detect signal over time. (SNR): Clarity of gnal over any nonspecific background. Densitometry is the quantitative measurement of optical density in light-sensitive materials, Such as photographic paper or photographie film, due o exposure to light.Optical density is a result of the darkness of a developed picture and can be expressed absolutely as the number of dark spots (i.e, silver grains in developed films) in a given area, i ; carom needed but usually it is a relative value, expressed in a scale.!™ Since density is usually measured by the decrease in the amount of light which shines through a transparent film, it is also called absorptiometry, the measure of light absorption through the medium. The corresponding measuring device is called a densitometer (absorptiomeier). The decadic (base-10) logarithm of the reciprocal of the transmittance is called the absorbance or density." PMax and DMin refer to the maximum and minimum density that can be produced by the material. The difference between the two is the density range.!"| The density range is related to the exposure range (dynamic range), which is the range of light intensity that is represented by the recording, via the Hurter~Driffield curve, In the context of photography, the dynamic range is often measured in "stops" . which is the binary logarithm of the ratio of highest and lowest distinguishable exposures; in an engineering context, the dynamic range is usually given by its decadic logarithm expressed in decibels, avion needec} According to the principle of operation of the densitometer, one can have: Spot densitometry: the value of light absorption is measured at a single spot line densitometry: the values of successive spots along a dimension are expressed as a graph bidimensional densitometry: the values of light absorption are expressed as a 2D synthetic image, usually using false-color shading