Academic literature on the topic 'Spectrophotometer. Absorption spectra'

Light is a form of electromagnetic radiation, usually a mixture of waves having different wavelengths. The wavelength of light, expressed by the symbol λ, is defined as the distance between two crests (or troughs) of a wave, measured in the direction of its progression. The unit used is the nanometre (nm, 10-9 m). Light that the human eye can sense is called visible light. Each colour that we perceive corresponds to a certain wavelength band in the 400-700 nm region. Spectrophotometry in its biochemical applications is generally concerned with the ultraviolet (UV, 185-400 nm), visible (400-700 nm) and infrared (700-15 000 nm) regions of the electromagnetic radiation spectrum, the former two being most common in laboratory practice. The wavelength of light is inversely related to its energy (E), according to the equation: . . . E = ch/ λ . . . where c denotes the speed of light, and h is Planck’s constant. UV radiation, therefore, has greater energy than the visible, and visible radiation has greater energy than the infrared. Light of certain wavelengths can be selectively absorbed by a substance according to its molecular structure. Absorption of light energy occurs when the incident photon carries energy equal to the difference in energy between two allowed states of the valency electrons, the photon promoting the transition of an electron from the lower to the higher energy state. Thus biochemical spectrophotometry may be referred to as electronic absorption spectroscopy. The excited electrons afterwards lose energy by the process of heat radiation, and return to the initial ground state. An absorption spectrum is obtained by successively changing the wavelength of monochromatic light falling on the substance, and recording the change of light absorption. Spectra are presented by plotting the wavelengths (generally nm or μm) on the abscissa and the degree of absorption (transmittance or absorbance) on the ordinate. For more information on the theory of light absorption, see Brown (1) and Chapters 2, 3 and 4. The most widespread use of UV and visible spectroscopy in biochemistry is in the quantitative determination of absorbing species (chromophores), known as spectrophotometry.

Circular dichroism (CD) is the ideal technique for studying chiral molecules in solution. It is uniquely sensitive to the asymmetry of the system. These features make it particularly attractive for biological systems. CD is by definition the difference in absorption, A, of left and right circularly polarized light (CPL): . . . CD = Ae − Ar . . . . . . 1 . . . CPL has the electric field vector of the electromagnetic radiation retaining constant magnitude in time but tracing out a helix about the propagation direction. Following the optics convention we take the tip of the electric field vector of right CPL to trace out a right-handed helix in space at any instant of time (1, 2). CD spectra can in principle be measured with any frequency of electromagnetic radiation. In practice, most CD spectroscopy involves the ultraviolet-visible (UV-visible) regions of the spectrum and electronic transitions, though increasing progress is being made with measuring the CD spectra of vibrational transitions using infrared radiation. We shall limit our consideration to electronic CD spectroscopy since the practical considerations for vibrational CD differ from those for electronic CD. For randomly oriented samples, such as solutions, a net CD signal will only be observed for chiral molecules (ones that cannot be superposed on their mirror images (3)). Oriented samples of achiral molecules, such as crystals, will also give a CD spectrum unless the optical axis of the sample aligns with the propagation direction of the radiation. However, such spectra are seldom useful. CD is now a routine tool in many laboratories. The most common applications include proving that a chiral molecule has indeed been synthesized or resolved into pure enantiomers and probing the structure of biological macromolecules, in particular determining the α-helical content of proteins. Figure 3 gives an example of a CD spectrum. The key points to remember are that a CD signal is observed only at wavelengths where the sample absorbs radiation, i.e. under absorption bands, and the signal may be positive or negative depending on the handedness of the molecules in the sample and the transition being studied.

Fluorescence spectrometry is the most extensively used optical spectroscopic method in analytical measurement and scientific investigation. During the past five years more than 60000 scientific articles have been published in which fluorescence spectroscopy has been used. The large number of applications ranges from the analytical determination of trace metals in the environment to pH measurements in whole cells under physiological conditions. In the scientific research laboratory, fluorescence spectroscopy is being used or applied to study the fundamental physical processes of molecules; structure-function relationships and interactions of biomolecules such as proteins and nucleic acids; structures and activity within whole cells using such instrumentation as confocal microscopy; and DNA sequencing in genomic characterization. In analytical applications the use of fluorescence is dominant in clinical laboratories where fluorescence immunoassays have largely replaced radioimmunoassay techniques. There are two main reasons for this extensive use of fluorescence spectroscopy. Foremost is the high level of sensitivity and wide dynamic range that can be achieved. There are a large number of laboratories that have reported single molecule detection. Secondly, the instrumentation required is convenient and for most purposes can be purchased at a modest cost. While improvements and advances continue to be reported fluorescence instrumentation has reached a high level of maturity. A review of the physical principles of the fluorescence phenomenon permits one to understand the origins of the information content that fluorescence measurements can provide. A molecule absorbs electromagnetic radiation through a quantum mechanical process where the molecule is transformed from a ‘ground’ state to an ‘excited’ state. The energy of the absorbed photon of light corresponds to the energy difference between these two states. In the case of light in the ultraviolet and visible spectral range of 200 nm to 800 nm that corresponds to energies of 143 to 35.8 kcal mol-1. The absorption of light results in an electronic transition in the atom or molecule. In atoms this involves the promotion of an electron from an outer shell orbital to an empty orbital of higher energy.

Malignant tumors of the orbit are the main cause for 41–45.9% of orbital tumor, and they will threaten both the organ of vision and the life of the patient. In our opinion, improving the effectiveness of treatment of malignant tumors can be implemented in the following areas: a) immobilization of doxorubicin in synthetic polymeric materials, which will fill the tissue structures that were resected and reduce the percentage of tumor recurrence. b) the use of nanomaterials for the delivery of doxorubicin to tumor cells. To develop a hydrogel implant and nanoparticles, to study the diffusion kinetics of doxorubicin in a hydrogel implant and the ability of nanoparticles to transport doxorubicin. The developed gels based on acrylic acid (AAc) were obtained by radical polymerization of an aqueous solution of monomers (AAc and N, N-methylenebisacrylamide (MBA)) at a temperature of 70°C. Matrices based on polyvinyl formal (PVF) were obtained by treatment of polyvinyl alcohol (PVA) with formaldehyde in the presence of a strong acid. Experimental studies were performed on rabbits of the Chinchilla breed, weighing 2–3 kg, aged 5–6 months, which during the study were in the same conditions. We implanted the hybrid gel in the scleral sac; orbital tissue and in the ear tissue of rabbits: Evaluation of the response of soft tissues and bone structures to implant materials was carried out on the basis of analysis of changes in clinical and pathomorphological parameters was performed after 10, 30 and 60 days. Diffusion of doxorubicin was examined by using UV spectroscopy [spectrophotometer-fluorimeter DS-11 FX + (DeNovix, USA)], analyzing samples at regular intervals during the day at a temperature of 25° C. The concentration of active substances was determined by the normalized peak absorption of doxorubicin at 480 nm. The release kinetics of the antitumor drug doxorubicin were investigated by using a UV spectrometer “Specord M 40” (maximum absorption 480 nm). The developed hydrogel implant has good biocompatibility and germination of surrounding tissues in the structure of the implant, as well as the formation of a massive fibrous capsule around it. An important advantage of the implant is also the lack of its tendency to resorption. Moreover, the results showed that the diffusion kinetics of doxorubicin from a liquid-crosslinked hydrogel reaches a minimum therapeutic level within a few minutes, while in the case of a tightly crosslinked - after a few hours. It was also found that the liquid-crosslinked hydrogel adsorbs twice as much as the cytostatic - doxorubicin. The analysis of the research results approved that the size of the nanoparticles is the main factor for improving drug delevary and penetration. Thus, nanoparticles with a diameter of less than 200 nm can penetrate into cells and are not removed from the circulatory system by macrophages, thereby prolonging their circulation in the body. About 10 nm. The developed hybrid hydrogel compositions have high mechanical strength, porosity, which provides 100% penetration of doxorubicin into experimental animal tissues. It was found that the kinetics of diffusion of drugs from liquid-crosslinked hydrogel reaches a minimum therapeutic level within a few minutes, whereas in the case of densely crosslinked hydrogel diffusion begins with a delay of several hours and the amount of drug released at equilibrium reaches much lower values (20–25%). The obtained preliminary experimental results allow us to conclude that our developed pathways for the delivery of drugs, in particular, doxorubicin to tumor cells will increase the effectiveness of antitumor therapy.

The purpose of this study is to measure the fluorescence properties of Indocyaninegreen (ICG) which is a fluorescence dye to be used as a fluorescence probe for the use of fluorescence imaging in biomedical applications. The fluorescence molecular imaging is expected to solve the issues in preclinical studies which require a lot of time, labors and sacrificed animals. Information of living body can be obtained by measuring the fluorescent properties of the probe in biological media. The absorption and emission spectra and the lifetime of ICG in non-scattering and scattering media were measured

In this study, we proposed and demonstrated an effective approach to model and predict spectral power distribution (SPD) for a phosphor-converted light emitting diode (pc-LED). For emission and excitation, broadband diffuse transmittances of 1 mm YAG:Ce phosphor plates with different concentrations were measured by a spectrophotometer. For emission, it was found that transmittance for all wavelengths was almost identical. This result indicates that emission spectrum prediction could be simplified by simulating the radiant power of the peak wavelength only. At the peak wavelength (560 nm), our

Solar energy utilization is very important in the background of global warming and reduction of carbon dioxide emission. Solar air/water collectors are convenient to convert solar energy into heat. Compared with regular solar collectors, direct absorption solar absorbers are not common. However, nanofluids can be used in direct solar absorbers to improve their performances. The objective of this paper is to evaluate the potential of application of nanofluids in direct solar absorbers. Spectral transmittances of nanofluids within the solar spectrum have been measured by a customized spectrophot