Validation Study of LuciPacTM A3 Surface for Hygiene Monitoring through Detection of ATP, ADP, and AMP from Stainless Steel Surfaces: AOAC Performance Tested MethodSM 051901

BACKGROUND: The LuciPac A3 Surface Hygiene Monitoring System based on the detection of total adenylate, ATP+ADP+AMP (A3), has been developed by Kikkoman Biochemifa. OBJECTIVE: This A3 swabbing assay kit was validated for Performance Tested MethodsSM (PTM) certification. METHODS: The LuciPac A3 Surface Hygiene Monitoring System was evaluated for limit of detection (LOD) for each adenylate with pure analyte solutions, detection of food residues and microbial residues on stainless steel surfaces, interference by disinfectants, and selectivity of the method response. RESULTS: Pure analyte studies performed by the method developer and the independent laboratory showed good linearity (R2 > 0.9854) and repeatability precision (RSDr < 20% for ≥2.5 fmol/assay). The LOD values for each adenylate were around 10 relative light units or 2.5 fmol/assay. The repeatability precision in the method developer laboratory for the matrix study (raw chicken breast, sliced deli ham, orange juice, yogurt, and apple pie) and the microbial study (Cronobacter sakazakii, Lactobacillus acidophilus, and Saccharomyces cerevisiae) were 8-30% and 10-35%, respectively. The repeatability precision of independent laboratory testing was 13-27% for orange juice and 16-43% for ham. Interference by several disinfectants indicate that rinsing is recommended to be performed after the use of sanitizing agents and before testing with LuciPac A3 Surface. Selectivity testing revealed that no positive interference and no inhibition were caused by adenylate analogues. Instrument variation, lot-to-lot consistency, accelerated stability (30°C, 5 weeks) were confirmed, and the method was shown to be robust against shaking time. CONCLUSIONS: The LuciPac A3 Surface has been successfully validated. HIGHLIGHTS: This A3 swabbing assay kit was qualified for PTM certification No. 051901.