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Absorbances are measured at the, the location of most absorbance in a molecule. Cells used for these kinds of spectrophotometer are optically matched cells, meaning the absorption of the cells are very much close to each other.Īfter the preparation of the standard and the sample solutions, each sample was then subjected to UV-Vis Spectrophotometry, measuring the absorbances for each sample. The PerkinElmer Lambda 35 UV-Visible Spectrophotometer is a double beam spectrophotometer – which means it makes use of two cuvettes or cells: one is for the reference, and the other for the sample. Some of the factors that affect the absorption characteristics of compounds are the following (Skoog,et.al., 2014): 1) Solvent, 2) Sample concentration, 3) Sample pH and 4) Sample Temperature. Also, pH and polarity may affect the absorption of some compounds, thus narrowing the choice of solvents for UV Spectroscopy. It involves measuring the amount of UV or visible radiation absorbed by a particular substance, Instruments that measure the ratio of the intensity of two beams of light in the UV-visible region are called UV-Visible Spectrophotometers (Behera,S., et.al., 2012)įor this determination, Methanol was used as a diluent, as Methanol has a cut-off wavelength in the UV range, thus making it a useful diluents for this particular experiment. UV- Visible Spectrophotometry is used for both qualitative and quantitative determinations of many compounds. The computations were made upon gathering all the necessary data from the UV-Vis Spectrophotometric determinations. Data was then recorded, as was treated statistically for the computation of the %Label Claim of the sample. Absorbances for each replicate was then measured by generating a scan for the standard first, then the sample. Using the 100ml Paracetamol Standard solution, the UV-Vis spectra for the standard was generated by setting the wavelength selection to 200 – 400nm. From this average weight, one sample and two replicates were weighed out, and the replicates were prepared using the same procedure as mentioned above.Ģ.4UV-Visible Spectrophotometric DeterminationĪ blank scan for the UV-Vis Spectra was first done, using water as the solvent.
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The Samples were prepared by weighing 20 paracetamol tablets and obtaining their average weight. One (1) ml aliquot was then obtained from this solution and was diluted further in a 100ml volumetric flask. The resulting solution was then diluted in a 250ml volumetric flask. Standard solution for paracetamol was prepared by weighing 500mg of the powdered Paracetamol standard, and dissolving it with 5ml of Methanol. 2.2Instrumentation Setupįor the instrumentation setup, the PC, as well as the UV-Vis Spectrophotometer was turned on – allowing the latter to warm up for at least 20mins. USP Grade Paracetamol was used in the determination as the Standard, and twenty (20) tablets of Biogesic® were obtained to be the sample paracetamol in tablet dosage form.
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UV-Visible Double beam Spectrophotometer, Perkin Elmer Lambda 35 with a spectral bandwidth of, wavelength accuracy of and a pair of matched quartz cells was used. Use the number format in separating procedures. This experiment focused on the determination of the Paracetamol content of Biogesic®, and to assess the Percentage Label Claim of the branded Paracetamol Tablet, in comparison to a USP Grade Paracetamol Standard. Based on the United States Pharmacopeial Convention, Titrimetric, Thin Layer Chromatographic Identification Test, as well as UV spectrophotometric assay determinations have been suggested as determination methods for Paracetamol in tablet formations.įor Paracetamol, various methods have been made known in its determination in bulk form, which is based on its reaction with Iron (III) and Ferricyanide in an HCl medium (Nagedra, P., 2010), or through a simultaneous determination of Paracetamol with another component, like Etodolac (Balan, P., et.al, 2011), Thiocolchicoside and Aceclofenac (Hapse, S.A., et.al, 2011) and Tramadol HCl ( Sawant, R.,et.al., 2011). Paracetamol,(N-(4-hydroxyphenyl) acetamide) is classified as to being part of the drugs known as “aniline analgesics” (Behera,S.,et.al, 2012) and is widely used as analgesic and antipyretic drug, together with caffeine, ibuprofen, and diclofenac sodium. Some of its advantages for Quantitative purposes arise from its wide applicability for analysis of compounds, and its apparent speed of performing analyses, with the use of modern technology applied to these instruments (Skoog, et.al, 2014). And this method has been very useful both for Qualitative and Quantitative Applications, especially for organic compounds. UV-Visible Spectrophotometry is a widely-used technique in Chemical and Pharmaceutical Analysis.