OPTICAL ACTIVITY

Valencia, Rey-Ann; Veloria, Evana; Vergara, James; Villena, John Mark; Walde, Rikkamae, Zamora, Ron Group 9 3A-Biochemistry Chem401 Laboratory
Optical Activity is a property of certain organic compounds, in this case sugars and amino acids, to rotate linearly or plane-polarized light due to the presence of chiral centers in their structures. The Optical activities of prepared solutions of cysteine and sucrose were observed and quantified using a Polarimeter, where a tube containing the sample is hit with a beam of polarized light, in the experiment water was used as the blank. 3 trials were allotted to ensure precision of data and after which values obtained for the blank was 0, -6.67 for the cysteine solution and 52.33 for the sucrose solution. Optical rotation was also computed from the observed rotation using an equation for Optical rotation allowing us to conclude that both samples are optically active but at different degrees and at different directions with sucrose being more optically active than cysteine.

ABSTRACT

INTRODUCTION
Optical activity is an intrinsic property of the molecular structure of organic compounds in which certain organic compounds have the ability to rotate the orientation of linearly or plane polarized light. This certain ability primarily arises as a consequence of the tetrahedral stereochemistry of sp3-hybridized carbon atoms also known as chiral centers wherein four different substituents are covalently bonded to it and the entire molecule with a chiral center is not identical to its mirror image. The relationship of the chiral molecule to its mirror image is much like the relationship of our right and left hands.

palms are facing the same way and placed over one another. Optical isomers also have no axis of symmetry, which means that there is no line that bisects the compound such that the left half is a mirror image of the right half. Thus the molecular groups that contribute most directly to optical activity are those that have mobile electrons which can interact with light. The relationship between optical activity and molecular structure results from the interaction of polarized light with electrons in the molecule. Generally, enantiomers have identical physical properties, such as densities, boiling points, melting points, among others. This poses a problem in the observance and quantification of enantiomerism. Fortunately, there is one physical property in which enantiomers differ: their ability to rotate plane-polarized light. The angle of rotation can be measured with an instrument called a polatimeter.

Figure 1 Exhibition of a carbon chiral center and its relationship to its mirror image.

If a compounds structure contains at least 1 chiral center, like the one shown in the image above, it and its mirror image are known as Optical isomers, or enantiomers, they have the same sequence of atoms and bonds but are different in their 3D shape. Two enantiomers are non-superimposible mirror images of one another (they are said to chiral), with the most common cited example being our hands. Our left hand is a mirror image of our right, yet there is no way our left thumb can be over our right thumb if our

Figure 2 Skematic Diagram of a polarimeter.

Ordinary white light consists of waves vibrating in all planes perpendicular to the direction in which the waves travel. Certain materials, such as a Nicol prism or the lenses of ordinary polaroid sunglasses, selectively transmit light waves vibrating in only one specific plane. This light is

said to be "plane-polarized". If another polarizing material is put in the path of the plane-polarized light so that its polarizing axis is parallel to the axis of the plane-polarized light then a maximum intensity of light will pass through. However, if the axis of the second polarizing material is perpendicular to the axis of polarized light then a minimum of polarized light will be allowed to pass through. Upon observance of the quantified rotation, calculations can now be made from the observed value of angular rotation by plugging the values. In solution, the formula for specific

rotation is:

Figure 3 Equation for Optical Activity

where is the observed rotation, l is the path length in decimeters, c is the concentration in g/ml, and [] is the specific rotation in degrees cm3 dm-1 g-1. An enantiomer that rotates plane-polarized light in the positive direction, or clockwise, is called dextrorotary [(+), or d-], while the enantiomer that rotates the light in the negative direction, or counterclockwise, is called levorotary [(-), or l-]. When both d- and l- isomers are present in equal amounts, the mixture is called a racemic mixture. Sucrose is a disaccharide containing a glucose and fructose monosaccharides that are both 6carbon containing sugars and capable of producing many optical isomers due to presence of 9 chiral centers. The presence of this large number of chiral centers indicates that more carbons contain a hydroxyl group pointing downward and in right direction favoring the light travel of the polarized light in the clockwise direction. Thus, sucrose was denoted as +, or dextrorotatory isomer. In contrast with cysteine containing only 1 chiral carbon has a negative angular rotation. This optical isomerism suggests based from the general formula of amino acids that has an L-configuration in which the light travel of polarized light will be counter-clockwise from the most favorable ionizable group, the CO2to the least or non-reactive substituent which is the hydrogen atom. This experiment aims to acquaint us with the use of the polarimeter in measuring optical activity and to compare the optical activity exhibited by two samples-sucrose and cysteine against water.

A. Sample preparation 5.00g of sucrose and 1.00g of cysteine were accurately weighed using an analytical balance and were placed in 2 separate Erlenmeyer flasks, both in solid form, solutions of each were prepared by dissolving the solid samples in distilled water with constant stirring to ensure complete dissolution, once dissolved, both were diluted to 50.00mL final volume. Theoretical molar concentration of both solutions were then computed and taken note of. B. Determination of Angular rotation. Once the solutions were prepared, angular rotation was then determined by the use of the Polarimeter. The polarimeter was first checked by turning it on and rotating the analyzer until 2 distinct fields-one dark, one light appeared in equal and crisp intensity. The polarimeter tube was then rinsed with distilled water in a wash bottle. It was then ready for reading. The polarimeter was calibrated with water as the blank by filing the tube with enough water to the top and capping in the stopper, air bubbles were avoided by making sure the tube is properly filled up and screwed capped not too tightly since the lenses exhibit double refraction under pressure. The glasses were then cleaned and the bubble was navigated to the bubble catcher of the tube for it not to interfere with the reading. The tube was then placed in the polarimeter and the analyzer was then rotated until the field seen through the eyepiece is equally illuminated. The reading was taken note of. After reading the blank was returned to its beaker and the tube was cleaned with distilled water in a wash bottle, replaced with cysteine solution following the same procedure and then replaced with sucrose solution. This entire procedure was repeated for 3 trials to ensure result precision. All readings were taken note of.

RESULTS AND DISCUSSION

EXPERIMENTAL PROCEDURES

Calculations for Optical Activity: Optical rotation = ____ CxL Sucrose ____52.33 = +254.52 cm3 dm-1 g-1 3 (0.1g/ cm ) (2.056dm) Cysteine ____-6.67 = -162.21 cm3 dm-1 g-1 3 (0.02g/ cm ) (2.056dm
Table 1 Obtained Angular Rotation and Computed specific Rotation of Cysteine and Sucrose

is with the results obtained from water-no chiral center thus is not optically active proven by the 0 degrees observed angular rotation in all 3 trials. Sucrose yielded a greater optical activity and is dextrorotatory as indicated by the + sign in the obtained results, as for cysteine it yielded a lower optical activity and is levorotatory as indicated by the sign in the obtained results and is further proven as the label in the reagent bottle showed the utilized cysteine sample is L-cysteine. The disaccharide Sucrose, with 9 chiral centers which are capable of producing 18 stereoisomers theoretically must have higher angular rotation and optical activity as compared to the amino acid Cysteine which only contains 1 chiral center, this theoretical fact was consistently proven by the obtained results in angular rotation values and calculated specific rotation values. Further more, it is safe to conclude that the number of chiral centers in a given molecule directly affects its degree of optical activity, more chiral centers means more substituents of the centers to be excited when hit by polarized light which in turn provides a denser electron cloud to interact with the photons and rotate the orientation of the plane-polarized light, thus possibly producing a more definite angular rotation value. The parameter of concentration affecting optical activity was not well proven by the experiment as we were only instructed to prepare 2 different solutions of 2 samples with 2 different concentrations. It could be established if only we were instructed to prepare solutions of the same sample but of varying concentration.

Reading in

Blank Water 0 0 0 0

Cysteine 0.165M -6.0 -7.0 -7.0 -6.67 -162.21 cm3 dm-1 g-1

Sucrose 0.280M +53.0 +52.0 +52.0 +52.33 +254.52

1 2 3 MEAN SPECIFIC ROTATION

cm3 dm-1 g-1

Considering the structures of the two samples Cysteine and Sucrose,

REFERENCES:
From books -McMurry, J. (2010). Foundations of Organic Chemistry, Cengage Learning Asia Pte. Ltd. Pp. 204210. -Laboratory Manual in Physical Chemistry, Department of Biochemistry, Faculty of Pharmacy, University of Santo Tomas as compiled by RVR. De Grano and MD Cation, Pp 28-29 - Farrington, D., & et.al. (1962). Experimenta Physical Chemistry. Wisconsin: McGraw-Hill. -Fredrick, H.G. (2008). Laboratory Exercises of Physical Chemistry. NY: John Wiley & Sons. From Internet -Natt, N.J. Zhu, A. (2011). ChemWiki: The Dynamic Chemistry Textbook Retrieved from http://chemwiki.ucdavis.edu/Organic_Chemistry/Chiral ity/Optical_Activity http://chemed.chem.purdue.edu/genchem/topicreview /bp/1organic/chirality.html

Figure 4 Skeletal formula of Sucrose

Figure 5 Skeletal formula of Cysteine

Chiral centers could be found on the structures of the samples, though 2 different type of organic compounds, both exhibit optical activity but at different degrees and different directions. These results conform to the theoretical assumption that they really are optically active compounds so