The Employment of Real-Time Polymerase Chain Reaction for the Identification of Bovine Gelatin in Gummy Candy

• Main Authors: Salamah, Nina, Erwanto, Yuny, Martono, Sudibyo, Rohman, Abdul
• Format: Article PeerReviewed
• Language: English
• Published: Universitas Gadjah Mada - Faculty of Pharmacy 2022
• Subjects: Pharmaceutical Sciences
• Online Access: https://repository.ugm.ac.id/283670/1/The%20%20Employment%20%20of.pdf; https://repository.ugm.ac.id/283670/; https://www.scopus.com/inward/record.uri?eid=2-s2.0-85140257188&doi=10.22146%2fijp.1970&partnerID=40&md5=76f66046ab42e24d44189327010a4dac

Gelatin is widely used in food products, especially candy. Gelatin can be made by hydrolysis of skin and bone of bovine and pig, and the obtained gelatins are typically called as bovine gelatin (BG) and porcine gelatin (PG). The use of PG in food and pharmaceutical products is prohibited to muslim consumers, therefore, the availability of analytical method capable of distinguishing BG from PG is very urgent. This study aimed to identify BG in gummy candy products using specific primers with real-time polymerase chain reaction (RT-PCR) methods. The specific DNA primer was designed using PRIMERQUEST and NCBI software and subjected to primer-basic local alignment search tool (BLAST). The assessment of primer specificity has been carried out using DNAs extracted from PG and BG and those extracted from raw meats of pigs, rats, dogs, goats, cows, and chickens. RT-PCR method using primer targeting on mitochondrial Cytochrome B gene was applied to analyze candies purchased from the markets. The sensitivity test was performed by measuring amplification at six dilution series (10000, 5000, 1000, 500, 50, and 10 pg/µL) on BG-based candies. RT PCR using specific primer could detect DNAs within the samples containing BG at an optimum temperature of 55.2°C. The DNA detection limit was 500 pg/µL. The standard curve of the serial dilution of DNA isolate of BG produces the correlation coefficient (R-value) of 0.866 and an efficiency value (E) of 996.2. Four products from the markets were examined using the designed primer. There from four samples were positive to contain BG. Real-time PCR using specific primer targeting on cytochrome-B (forward: 5’-ACTAGCCCTAGCCTTCTCTATC- 3’ and reverse: 5’ - TGTCAGTAGGTCTGCTACTAGG-3’) can be used for halal authentication analysis of gummy candy. Copyright © 2022 by Indonesian Journal of Pharmacy (IJP).