Editor-in-Chief Hatice Kübra Elçioğlu Vice Editors Levent Kabasakal Esra Tatar Online ISSN 2630-6344 Publisher Marmara University Frequency Bimonthly (Six issues / year) Abbreviation J.Res.Pharm. Former Name Marmara Pharmaceutical Journal
Journal of Research in Pharmacy Articles in Press
Optimisation and validation of end-point PCR for the detection of porcine DNA in anti-ageing cream products containing collagen
Novi YANTIH1,Era WIDIANINGSIH2,Sri SURATI2,Andi ASNAYANTI2
1Faculty of Pharmacy, Universitas Pancasila, Jakarta, Indonesia
2Laboratory of Microbiology and Molecular Biology, National Quality Control Laboratory of Drug and Food, National Agency of Drug and Food, Jakarta, Indonesia
DOI : 10.29228/jrp.2022.00 Collagen is the protein-building block of the skin, muscles, bones, tendons, ligaments, and other connective tissues. It is commonly used as an anti-ageing agent in various cosmetic products, particularly anti-ageing creams. The most predominant collagen in the market is derived from pigs and cows. Based on Indonesian Law No. 33 of 2014 governing the halalness of products marketed in Indonesia, there is an emerging need to determine scientifically the halalness of products. Cosmetics containing porcine and its derivatives are classified as non-halal products. Therefore, there should be a robust method for detecting porcine-derived collagen content in anti-ageing creams. Polymerase chain reaction (PCR) can be an effective method for determining the animal source of products through DNA detection. This study aimed to determine the optimum DNA isolation and extraction conditions for the PCR detection of porcine DNA fragments. Incubation was performed twice for 30 min at 60°C and for each incubation, 5 mL of lysis buffer, and 25 L of proteinase K were used. Amplification was performed for 45 cycles and electrophoresis was conducted for 60 min at a voltage of 120 volts and a current of 100 mA. Validating the detection of porcine DNA in anti-ageing cream containing collagen using PCR resulted in a specific and robust method for detecting porcine DNA with a detection limit of 0.004 ng/μL. The repeatability of 10 repetitions consistently showed a band of 149 bp, with 0% false-positives and false-negatives. This method is valid for detecting pig DNA in anti-ageing creams containing collagen. Keywords : validation ; PCR, collagen; porcine DNA; anti-ageing cream
Marmara University