1318895107950 Polarimeter verification and validation Name of student

1318895107950
Polarimeter verification and validation
Name of student: Lim Si Hui Perris
Administration Number: 1605742D
Care Group: L2
Name of Supervisor: Alistair Chan
Company: Leung Kai Fook Medical Co Pte Ltd
A report submitted for the partial fulfilment of the
requirement for the
Diploma in Pharmaceutical Science
019
Contents
Contents
Chapter 1: Introduction
Chapter 2: Literature Review
Chapter 3: Material and methods
Material
Methods
Setting up of polarimeter instrument
Preparation of Menthol and Camphor standards solution
Preparation of sucrose standards solution
Water zero verification of polarimeter analysis
Zero Verification of 96% ethanol
Sample analysis
Chapter 4: Time chart or Gantt chart
Chapter 5: Reference
Introduction
Optical rotation is an important property especially to most pharmaceutical Industry. Most pharmaceutical substance are optically active where they rotate at an incident plane of polarized light so that the transmitted light emerged at a measurable angle. Polarimeter, the measurement of optical rotation is then convenient used to distinguish isomers with chiral indication and its purity.

Substance that shows optical rotatory properties are chiral. Those that rotated in clockwise direction towards the light source are known as Dextrorotatory or (+) optical isomers. Likewise, those polarized light rotating in an opposite direction, anti-clockwise direction is known as levoratatory or (-) optical isomers.

Validation is a confirmation by examination and a provision objective evidence to prove that the instrument is fit for its intended used. Hence it is a very important procedure before any actual analysis. In the current principle of polarimeter validation, sucrose solution is commonly used as a standard to validate the polarimeter instrument however this method does not prove that the instrument is accurate as it is a one-point validation. Thus, a three-point validation is proposed with additional standards such as the use of menthol and camphor standards. Three-point validation was proposed because it is to create a linearity range which will be able to test the robustness characteristic of the optical rotation, allowing the results obtained from routine sample to be more reliable.

This project aims to ensure that the performance of polarimeter instrument is fit for its intended use when performing routine analysis of raw materials. This project will be evaluated by using menthol and camphor reference standard purchased from Sigma Aldrich to validate the performance of polarimeter. It will be done by weighing a quantified and dilute an amount of standard using a suitable solvent to achieve a desired concentration.

Literature Review
Polarimeter instrument
Background
The polarimeter instrument measure the optically active substances in a solution. This technique used is called the polarimetry. Polarimetry is based on the bending of polarized light as it passes through the optically active solution. When the light passes through the polarizer a polarized light will then be formed and if the angle of polarized light has been changed, it indicates the presence of optically active substance. The degree of polarized light is proportional to the concentration of optically active substance.
In polarimetry, substance that rotates the polarized light to the left are known as levorotatory. On the other hand, substances that rotates polarized light to the right are known as the dextrorotatory.
In a specific rotation of a sugar solution at 20oc and a wavelength of 589 nanometres, the optical rotation of sucrose is +66.5o.
Factors affecting polarimetry
In polarimetry, such factors like temperature of sample, Wavelength, Length of polarimeter tube, nature of non-sucrose substance in the sample and the concentration of optically active substances in the sample affects the degree of polarization.

Hence in most sugar labs, the temperature, wave length and the length of polarimeter tube were kept constant.
Calibration of polarimeter
In the International Commission for uniform Methods of Sugar Analysis (ICUMSA) methods, the recommended conditions for calibration is to use normal sucrose solution where 26.00 grams of pure sucrose were being diluted in distilled water to a final volume of 100 millilitres and the standards temperature should maintained at 20oc throughout the whole calibration analysis. The wavelength of the polarimeter should be also set to 589 nanometer.
Hence, under such conditions the optical rotation of sucrose solution in a 200 millimeter polarimeter tube a 20oc is 66.54. like the 100 scale of polarimeter.
Formula for polarimeter calculation
When using different weight or polarimeter tube size the following equation when stated below was being used to determine the polarimetric reading at normal weight
Equation:
% Sugar = Polarimetric reading x (26.0 /Mass Sample) x (200/Tube length) x (Flask Volume / 100)
Calibration of Polarimeter instrument using sucrose solution
Background
Pure solution of sucrose diluted in water was being used to calibrate the polarimeter instrument however a strict protocol is required to ensure accuracy.

In Bellingham + Stanley (B + S) polarimeter, they are equipped with International Sugar Scale (ISS) or Z Scale. In the 100 oZ point of ISS scale is known as optical rotation of pure sucrose in distilled water (26.00g in 100ml of distilled water.)
Thus, it can be used to calibrate the polarimeter instrument.

Problems in preparation of sugar solution
Volumetric method is more prone error as volumetric flask may be not accurate and a strict temperature control is required to ensure that liquid density is accurate.
In addition, when a substance dissolve in another substance, an exothermic and endothermic reaction may occur which led to changes in temperature during mixing. Thus, the accuracy of concentration will be affected.
ICUMSA Method
ICUMSA also known as International Commission for uniform Methods of Sugar Analysis is an international recognised and approve method use in sugar industry. Where it requires strict protocol for preparation of sugar solution.
Bellingham and Stanley
In B+S method, it was recommended that sucrose solution was not recommend to be used as a standard solution to calibrate the polarimeter instrument due to the errors that might occurs in sample preparation of sucrose solution and when the sample degrades, the optical rotation value will be affected thus fresh sucrose solution is recommend to used to calibrated the polarimeter.

For accurate calibration, B+S methods recommend the used of Quartz Control Plates as the vales obtain from Quartz Control Plates can be trace thus is more reliable than sucrose solution standard. In addition, Quartz Control Plate are more stable, and it provides long term reference.

However, in Quarts control Plates, it still requires a good temperature control to ensure the accuracy thus the operator must ensure that the correct temperature was applied before the analysis.
Materials and Methods
Materials
Menthol and Camphor standards are obtained from Sigma Aldrich Pte Ltd. Pure Sucrose are obtained from Merck. 10ml Volumetric flask where used for dilution. Deionized water was used to dilute the sucrose standard while 96% ethanol where used to dilute menthol and camphor standard. All standards will be weigh using a Mettler Toledo Ltd Analytical weighing balance and will be grinded using a glass pestle and mortal.

Methods
Setting up of polarimeter instrument
Polarimeter instrument is switch on before the analysis for a few mins to allow instrument to be stabilized. An air zero is being perform before proceeding to select ‘set up’ button. When prompted for password, In put ‘123’. Select option ‘measurement’ > ‘Mode’ > select multiple. Once done, select option ‘operational parameter’ > ‘communication settings ; ‘file export option’ ; ‘file export location’ ; ‘folder’ ; ‘Browse folder’ ; ‘removable’ and select the respective folder to save in. Ensure that the time and date is accurate. If not, select ‘operational parameters menu’ ; ‘time and date’ to set the time and date.

Preparation of Menthol and Camphor standards solution
Using a mortar and pestle, camphor and menthol crystals were being grind into powders. 0.1g of menthol and camphor reference standard purchased from Sigma Aldrich were being weight into two 10ml volumetric flask using an analytical weighing balance and record the weight obtain in a logbook. Using a marker label a 10ml volumetric flask with the standard name, lot number, name of analyst, Concentration of the standard and the date of expiry. Top up with 96% ethanol to the calibrated mark and ensure that the solute is fully dissolve into the solvent by inverting the volumetric flask. Once the solute has been dissolved, place the solution into the refrigerator for three minutes.
Preparation of sucrose standards solution
0.1g of Sucrose were being weigh into a 10ml volumetric flask using an analytical weighing balance and record the weight obtain in a logbook. Using a marker label the 10ml volumetric flask with the standard name, lot number, name of analyst, Concentration of the standard and the date of expiry. Top up with deionized water to the calibrated mark and ensure that the solute is fully dissolve into the solvent by inverting the volumetric flask. Once the solute has been dissolved, place the solution into the refrigerator for three minutes.
Water zero verification of polarimeter analysis
Rinse the cell three times with deionized water and top up the cell with deionized water. Ensure that there is no air bubbles in the cell. Using a tissue, remove excess water on the external surface of the cell by using an dabbing motion. Place the filled cell into the polarimeter chamber, place the thermometer probe into the sample and close the chamber door. Select ‘Zero’ to zero away the cell and the deionized water.
Zero Verification of 96% ethanol
Rinse the cell three times with 96% ethanol and top up the cell with 96% ethanol. Ensure that there is no air bubbles in the cell. Using a tissue, remove excess ethanol on the external surface of the cell by using an dabbing motion. Place the filled cell into the polarimeter chamber, place the thermometer probe into the sample and close the chamber door. Select ‘Zero’ to zero away the ethanol.
Sample analysis
Rinse the cell three times with standard solution and top up with standard solution. Ensure that there is no air bubbles present in the cell. Using a tissue, remove excess solution present on the external surface of the cell by using a dabbing motion. Place the filled cell into the polarimeter chamber, place the thermometer probe into the solution and close the chamber door. Ensure that temperature is within 24 to 26. Before proceeding with analysis. Select ‘scale’ ; ‘Specific rotation’ ; ‘Solution’ ; under concentration, input weight of sample. Select start to run the analysis ; input lot identifier as sample name ; Input sample identifier as replicate number. Once done, wait for that results and print accordingly.

center43434000Gantt Chart
Reference