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Submitted: 07 February 2018 Modified: 03 August 2018
HERDIN Record #: R07-CIM-18020718010325

The effect of storage on refrigerated temperature on the antioxidant property of Citrofortunella microcarpa (Calamansi) using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay.

Gail Ann Bongalos,
Arvin Dela Vega,
Mark Gavin Dinsay,
Kenn Carlson Dy,
Mikaela Adeliz Gorme,
Bea Angeli  Kuizon,
Lalaine Mayol,
Esther Ann Nacita ,
Jane Marie Valmores,
Lenie May Vaporoso

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Introduction: Antioxidant compounds are the substances, compounds or nutrients found in food which can prevent or slow oxidative damages in the body. Antioxidants play an essential role against free radicals by neutralizing free radicals through donation by one of their electrons. Among the plant sources of antioxidant-rich compounds, citrus fruits like Citrofortunella microcarpa (Calamansi) are widely consumed by Filipinos. The antioxidant content of citrus fruits is affected by storage conditions, particularly temperature and method of preparation. Objective: This study aims to detect the decrease in the antioxidant property of Citrofotunella microcarpa (Calamansi) extracts after storage intervals of 24,48, 72 hours at 4°C (±0.5°C). It further aims to determine the absorbance values of the extracts and correlate these to the standard Vitamin C absorbance values, the extent of the decrease in absorbance of the mentioned storage intervals at 4°C (±0.5°C) of the extracts. Design and Setting/ Population: A quantitative-experimental study was used to determine the extent of absorbance of the different time intervals. The study was performed at the CIM Biochemistry and Histology Laboratory. The population included Citrofortunella microcarpa (Calamansi) fruits with green intact skin that are firm to touch. Fruits that were worm-infested, overripe (yellow to orange in color) with bruised and damaged skin and discolored with black blotches were excluded in the study. Methodology: Approximately one hundred Citrofortunella microcarpa (Calamansi) were used to collect 60 ml extract of Calamansi. The collected extract was further subdivided into 12 equal aliquots containing 5 ml each. The aliquots were divided into four groups for the 0, 24,  48 and 74-hour antioxidant activity assay. All vials except the 0-hour group were stored in a refrigerator set at 4°C (± 0.5 °C). Water was used as negative control - 60 ml was divided and stored under the same conditions as the test samples. Sodium ascorbate (vitamin C) preparation from a capsule (Fern-C), at an initial concentration of 0.25 mg/ml, was used as a positive control and a standard to derive the calibration curve. 2,2-diphenyl-1-picrylhydrazyl (DPPH) prepared by dissolving 10 mg in 250 ml of 80% ethanol, was used to determine the radical scavenging activity of each test group. Absorbance was measured at 517 nm. A quantitative decrease in antioxidant content was hypothesized to occur with increased storage duration. A line graph was made by plotting the time against the absorbance. The value reflects the change in antioxidant activity in relation to time. Results: The results showed no trend in the vitamin C standardization. The results showed that there was an increase in absorbance with increase duration of storage of calamansi extract. Negative control values from 0-72 hours had lower absorbance values than those of the test sample. Discussion: Since the expected trend of the vitamin C standardization curve was not obtained and the relation to negative control absorbance values was anomalous, the significance test results cannot be determined. The researchers postulated that different factors in the experiment could have affected the results. Conclusion: The test sample results cannot be evaluated because there was no linearity observed in the vitamin C standardization assay and the negative control values does not conform to the principles stated in the related literature. Recommendations have been made to improve the outcomes of future studies.

Publication Type
Thesis Degree
Publication Date
April 2016
LocationLocation CodeAvailable FormatAvailability
Cebu Institute of Medicine Abstract
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