Determination of Vitamin C content in the commercial fruit juices by utilizing high public presentation liquid chromatography ( HPLC ) and spectrophotometry method.
Presents, market is flooded with those fruit juices or sirup with the label “ vitamin C enriched ” . Vitamin C had become a celebrated compound in medical and nutrient industry. Then what is vitamin C and its map?
Vitamin C is an indispensable water-soluble vitamin and it is the first vitamin being discovered by the Magyar doctor Professor Albert Szent-Gyorgyi from the adrenal cerebral mantle of cowss. ( Davies et al. , 1991 ) . Vitamin C can be found in most of the veggies and fruits. To mensurate the Vitamin C in fruits, it is important to mensurate both the ascorbic acid and dehydroascorbic acid ( DHAA ) . This is because ascorbic acid is readily oxidized to DHAA and the oxidation procedure is reversible and DHAA is besides biological active as ascorbic acid. However, DHAA can be irreversible be converted to diketogulonic acid, which is non biologically active. ( Isabel et al. , 2007 )
Harmonizing to Institute of medical specialty ( IOM ) , the Dietary Reference Intakes ( DRIs ) for vitamin C for grownup work forces and adult females is 90 mg/day and 75 mg/day severally. ( Food and Nutrition Board, Institute of Medicine, 2004 ) . Lack of vitamin C cause scorbutus, which is a disease Characterized by shed blooding gums, impaired wound healing, anaemia, depression and weariness. ( Phillips et al, 2010 ) . So that, it is of import to do certain that we have adequate vitamin C consumption.
Vitamin C plays an of import function in human organic structure. It serve as extremist scavenger in vivo, this make it an of import antioxidant in our organic structure. Besides that, Vitamin C is besides a cofactor in assorted physiological procedures. Furthermore, it besides participates in immune system, biogenesis and metamorphosis of certain compounds.
Assorted methods had been adapted to find vitamin in many industry, which include spectrophotometry, titrimetry, voltammetry, fluorometry, potentiometry, kinetic-based chemiluminescence ( CL ) , flow injection analyses and chromatography. ( Arya et al, 2000 ) . Of class, each of the methods has their strong and failings.
Furthermore, due to vitamin C can oxidise easy, so the finding process has to be designed to avoid the loss of the vitamin. Factors including the temperature, visible radiation, pH and O exposure can impact the vitamin C content. ( Phillips et al, 2010 ) .
1. To find the vitamin C content in commercial fruit sirup.
2. To measure the feasibleness of utilizing HPLC method for finding vitamin C.
3. To compare the different vitamin C content in different trade name of commercial fruit sirup.
4. To compare the dependability of spectrophotometric and HPLC method in vitamin C finding.
2.0 Literature reappraisal
2.1 Discovery and construction of vitamin C
In the twelvemonth 1928, a Magyar doctor Professor Albert Szent-Gyorgyi was successfully to roll up a less sum of an whitish crystalline substance from the cerebral mantle of cowss. After the find of this whitish crystalline compound, he had done a series of trial on it. The consequence showed that, the substance can bleach I. Following, it besides lower the vapour force per unit area of the H2O, which gave the comparative molecular mass of the crystal about 180 g/mol. ( right value=176.4 g/mol ) . He besides did a burning analysis on the compound, which gave 40.7 % C, 4.7 % H and 54.6 % O. This information enable him to infer the C6H8O6 as the molecular expression for this compound. Subsequently, he named the compound as ‘hexuronic acid ‘ but eventually changed to ascorbic acid which average bar of scorbutus. ( Davies et al. , 1991 )
All in all, ascorbic acid is a six C substance with 4 OH group with comparative molecular mass of 176.14 g/mol. The thaw point is 190-192oC and the specific rotary motion is +23o in H2O and +49o in methyl alcohol. ( Davies et al. , 1991 ) .
2.2 Properties and functions of vitamin C
2.2.1 Redox properties- an effectual antioxidant
Figure ( Adapted from J. Verrax et Al. 2008 )
Due to the alone construction of vitamin degree Celsius ( figure 1 ) , it can be easilly be reversibly oxidised to dehydroascorbic acid or ascorbate and both of them can lost electro to organize ascorbyl extremist. ( Verrax et al. , 2008 ) . This makes vitamin C an of import antioxitant and cut downing agent involve in many physiological reactions such as, growing, reproduction and even unsusceptibility. ( Phillips et al, 2010 ; Wahli et Al, 2003 )
2.2.2 Role of vitamin C in forestalling diseases.
As the name ascorbic acid significance that forestalling scorbutus, it plays an of import function in forestalling scorbutus. Scurvy is the vitamin C lack disease which has the symptoms desertion of collagen, failure of lesion healing, anaemia and hemorrhage gums. ( Verrax et al. , 2008 ; Phillips et Al, 2010 ) . If abject remain untreated, it can be fatal.
Following, vitamin C besides involve in malignant neoplastic disease intervention. Ascorbate enhances the activity of hydroxylase ( inhibit HIF-1 ) by maintaining the Fe Centre of hydroxylases in a decreased province. Hypoxia-inducible factor 1 ( HIF-1 ) is a tumour bring oning agent. ( Verrax et al. , 2008 ) .
A research had been done by Wahli T. et. Al. upon the influence of dietetic vitamin C on the lesion mending procedure in hurt rainbow trout. They really feed them with different degree of vitamin C and compare the lesion mending procedure. The consequence had prove that, bulk of the rainbow trout provender with high degrees of vitamin C showed more rapid wound mending procedure. ( Wahli et al, 2003 ) .
2.2.3 Vitamin C composites and the map.
In certain status, ascorbic acid can deprotonate at 2nd and 3rd C OH group to organize monoanion and dianion. These two anion will so respond with metal to organize functional ligands monodentately or chelation.. One of the illustration is the composites with Fe which will better the Fe soaking up in bowel. ( Zumreoglu-karan, 2006 ) . Ascorbic acid can besides adhere to press Centre of hydroxylases to heighten its activity in malignant neoplastic disease intervention. ( Verrax et al. , 2008 ) .
3.0 Methodology Review
UV/visible spectrophotometer is one the most normally used instrument in analytical chemical science. UV/visible spectrophotometer provide a fast, simple and dependable method for finding. However, it can be interfered by other compound such as sugars and some organic acid. The light beginning of the UV/visible spectrophotometer can be switched to supply the maximal soaking up for the sample. The optimal soaking up for vitamin degree Celsius in impersonal solution is at 265nm.
To get down the experiment, standard solutions has to be prepared on the topographic point for standardization and the sample has to be diluted. Stabilizer can be added if it is needed to stabilise the vitamin C. Then by utilizing distilled H2O as a mention, record a spectrum of the highest concentration criterion to obtain the wavelength of the optimal soaking up. Last is to mensurate the optical density value of all the standard solutions and the sample. Then plot the standardization graph. After the consequence is obtain, SPSS is used to analyse the consequences. ( Chemistry and Biology division, TAR college, n.d. , p.1-10 )
3.2 high public presentation liquid chromatography ( HPLC )
High public presentation liquid chromatography ( HPLC ) forces the dissolver through the bantam column utilizing high force per unit area. This method allows smaller atoms to be used to supply a higher surface country and therefore increase the sensitiveness. Furthermore, all solute molecule will go in more unvarying due to little diameter of the column. This will cut down the grade of eddy diffusion and cut down the home base tallness.
To get down the experiment, standard solutions has to be prepared on the topographic point for standardization and the sample has to be diluted. Stabilizer can be added if it is needed. Then test the standard solution and vitamin C by utilizing the HPLC instrument. After the consequence is obtain, SPSS is used to analyse the consequences. ( Chemistry and Biology division, TAR college, n.d. , p.57-59 )