MOLECULAR DETECTION AND IDENTIFICATION OF ASPERGILLUS NIGER CONTAMINATION IN COSMETIC/PHARMACEUTICAL RAW MATERIALS AND FINISHED PRODUCTS

Abstract

A PCR‐based assay using a one step sample preparation was developed and validated against standard methods for the rapid detection of Aspergillus niger contamination in cosmetic/pharmaceutical raw materials and finished products. Artificially contaminated samples were added to Sabouraud Dextrose Broth (SDB) with 10% Tween 20 and 3% Soy Lecithin. Samples were then agitated at 200 revolutions per minute for 24 h at 35C. Prior to the PCR assay, sample preparation consisted of adding 10 μL of the 24 h enrichment broth sample mixture to a lysis buffer containing Tris‐EDTA and 0.4% SDS. DNA extraction was accomplished by boiling for 1 h to release the mycelial DNA. After DNA extraction, a 50 μL aliquot of the lysate was then added to a PCR tube containing the following ingredients: 4 Ready‐To‐Go PCR beads, 2 μL of two fungal primers, and 48 μL of sterile water. The DNA primers encoded for specific sequences of the Aspergillus spp. 18S rRNA gene. A 363 bp DNA fragment was detected in all of the artificially contaminated samples. Standard methods require 6–8 days to complete the isolation and identification of mold contamination. However, the PCR‐based assay was completed within 27–30 h. Rapid detection of mold contamination in cosmetic/pharmaceutical raw materials and finished products can be used to optimize the quality evaluation of a sample to allow the release in 27–30 h.

Microbiologically Contaminated and Over-Preserved Cosmetic Products According Rapex 2008–2014

“…Other studies have showed that the presence of B. cepacia in products used for mouth hygiene has frequently been implicated in B. cepacia infection [29][30][31]. Contamination of cosmetics and pharmaceuticals with B. cepacia is also a frequent problem in the United States and it is a cause of major product recalls [32][33][34]. B. cepacia can cause diseases primarily among immuno-compromised populations [35].…”

Microbiological risk assessment for personal care products

“…Similarly in the EU, recalls notified by the Rapid Alert System for non-food consumer products (RAPEX) between 2005 until week 17 in 2008 showed contamination by P. aeruginosa was also the most frequently identified microorganism in the 24 products recalled due to microbiological contamination [31]. Jimenez and Smalls [32] noted also that members of the genus Burkholderia, notably B. cepacia, are increasingly responsible for recall of products.…”

Ausbruch mit Burkholderia-cepacia-Komplex durch kontaminierte Mundspüllösung

“…Sie können bereits herstellerseitig kontaminiert sein und zu Ausbrüchen mit diesen Erregern füh-ren [4,15,20]. Dabei kommt auch Burkholderia-cepacia-Komplex (Bcc) als Kontaminant solcher Produkte in Betracht [13,17]. Dies sind gramnegative Bakterien, die v. a. in der Umwelt gefunden werden und als opportunistische Krankheitserreger vorkommen [17].…”