Optimisation and validation of end-point PCR for the detection of porcine DNA in anti-ageing cream products containing collagen

Abstract

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

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