The Ultimate Guide for Learners and Buyers Are you planning to buy a UV Vis Spectro photometer? We know that buying a UV Vis Spectro Photometer can be a tough job especially when you do not have expertise in it. Interesting thing is that – you are not alone. After helping 1000+ customers buy the right UV Vis Spectro Photometer, we have planned to make the most detailed guide on how to buy a UV Vis Spectro photometer without having technical knowledge. The guide is easy to read and follow whether you are in your early 20’s or 50+ age. Anyhow, if you still need our help in selecting the right model of Spectrophotometer, our technical team is available 24/7 for your help free. Which things did you need to know before buying a Spectrophotometer? A brief knowledge about UV Vis Spectro Photometer and its applications Your careful attention for 15 minutes to read this guide. Yes, only 15 minutes you need to understand, how to buy UV Vis Spectro Photometer and you will pass through the whole steps. In this guide, you will cover. Chapter 1: Spectrophotometer Overview Chapter 2: Working principle of Spectrophotometer Chapter 3: Types of Spectrophotometer Chapter 4: Main parts of Spectrophotometer Chapter 5: Troubleshooting of common problems, which occur during operation Chapter 6: Spectrophotometer applications Chapter 7: Merits and demerits of HPLC Chapter 8: Famous brands of HPLC If you want to Learn HPLC then please read the article Learn HPLC: The Ultimate Guide for Learners & Buyers Chapter 1 Spectrophotometer Overview Ultraviolet-visible (UV Vis) spectrophotometry is a quantitative technique used to measure light absorbance across the ultraviolet and visible ranges of the electromagnetic spectrum. When the incident light strikes on the matter it can either be absorbed, reflected, or transmitted. The atomic excitation occurs due to the absorbance of radiations in UV Vis range, which refers to the transition of molecules from the ground state to an excited state. Before an atom can change excitation states, it must absorb sufficient radiation for electrons to move into higher molecular orbits. If there is a Shorter bandgap between electrons then there will be the absorption of shorter wavelengths of light. A UV-Vis spectrophotometer measures the intensity of light transmitted through a sample compared to a reference measurement of the incident light source. The amount of light absorbed is directly proportional to the concentration of the sample and the distance the light travels through the sample. Therefore, UV-Vis spectrophotometers are able to determine the concentration of specific analytes in a microvolume by controlling the Wavelengths and path length. Chapter 2 Working principle of Spectrophotometer The working principle of the Spectrophotometer is based upon Beer-Lambert’s law, this law stated as the amount of light absorbed by a color solution is directly proportional to the concentration of the solution and the path length. Mathematically, A ∝ cl Where, A = Absorbance / Optical density of solution c = Concentration of solution l = Path length or, A = ∈cl ∈ = Absorption coefficient Chapter 3 Types of UV Vis Spectro photometer TYPES OF SPECTROPHOTOMETER Following are the two different types of spectrometer Single beam spectrophotometer Double beam spectrophotometer Single Beam Spectrophotometer: This type of spectrophotometer works between 325 nm to 1000nm wavelength using a single beam of light. Light travels in uni-direction and read the blank and test solution same. Double Beam Spectrophotometer: This type of spectrophotometer works between 185 nm to 1000 nm wavelength. This instrument splits the light from the Monochromator into two beams. One beam is used for reference and the other for sample reading. It eliminates the error which occurs due to fluctuations in the light output and the sensitivity of the detector. Chapter 4 Main parts of UV Vis Spectro photometer MAIN PARTS OF SPECTROPHOTOMETER There are following Seven different parts of spectrophotometer Light source: In spectrophotometer different sources of light are used to produce light of different wavelengths. Tungsten lamps common source of light used in the spectrophotometer for the visible spectrum. Hydrogen lamp and the deuterium lamp for Ultraviolet radiation. Nernst filament or globaris is the most satisfactory source of IR radiation. Monochromator: The main function of a monochromator is to separate the color components of light. To select the particular wavelength, prism or diffraction grating is used to split the light from the light source Sample holder: Test tubes or Cuvettes are used to hold the colored solutions. They are made up of glass at a visible wavelength. Beam splitter: It is present only in a double beam spectrophotometer. It is used to split the single beam of light coming from the light source into two beams. Mirror: It is also present only in a double beam spectrophotometer. It is used in the right direction to the split light from the beam splitter. Photodetector system: When light falls on the detector system, an electric current is generated that reflects the galvanometer reading. Measuring device: The current from the detector is fed to the galvanometer. The reading on the meter is directly proportional to the intensity of light. Chapter 5 Troubleshooting of common problems, which occur during operation Problem 1: When the instrument scans the sample, a straight line is displayedPossible cause: The software is malfunctioning.Solution: Exit the operating system, restart the computer, and scan again. Problem 2: After the UV/Visible spectrophotometer is turned on, the light source is not brightpossible reason:①The light source bulb is damaged;②The fuse is burned out.Solution: replace the deuterium lamp or tungsten lamp; replace the fuse. Problem 3: The instrument noise is relatively largePossible cause: The light source bulb has been used for longer than its life span.Solution: Replace the light source bulb. Problem 4: A certain section of the baseline is particularly noisyPossible cause: The corresponding filter in the