Hornback, M.1, Alburty, D.1, Long, B.1, Lewis, A.1 and Page, A.1, (1)InnovaPrep LLC, Drexel, MO, USA
Poster
Beer spoilage organisms and contamination present a major risk for the brewing industry. As such, microbiological testing for these organisms is necessary throughout the brewing process. However, most laboratories still use conventional cultivation methods, which are time-consuming—requiring 3 to 5 days for beer to be released to the market. Rapid microbiological analytical methods offer great potential for increasing the reliability of spoilage detection in beer while reducing labor costs and product hold times; however, small analysis volumes limit the usefulness of these methods. Furthermore, development of rapid detection methods has far outpaced development of sample collection and concentration techniques, which are necessary to enable detection of low microbial concentrations in the brewing process. The Concentrating Pipette is an automated, rapid bio-concentration device. Samplers are first filtered through high-flow, single-use pipette tips capturing microorganisms from the fluid sample matrix. Once the microorganisms are captured, the automated wet-foam elution process recovers and delivers the microorganisms into a microliter volume of clean buffer ready for analysis by modern or classical methods. In this study, InnovaPrep’s Concentrating Pipette was investigated as a bridge to concentrate 12 oz of beer into volumes more appropriate for rapid detection methods. The high level of carbonation in beer created a significant hurdle in applying the Concentrating Pipette to this application. During processing significant quantities of CO2 are released from the beer, causing the hydrophilic membrane filter to lock up. Although there are numerous decarbonation methods, none of these methods were able to reduce the amount of residual CO2 that allowed an entire can of beer to be processed by the Concentrating Pipette. A method was devised to create nucleation sites in glassware to allow for more efficient decarbonation. In short, 12 oz (355 mL) of room temperature American lager beer was poured into glass containers that were sandblasted to create a large surface area of nucleation points to help in the degassing process. The beer was incubated at 4°C for 10 min to increase the solubility of CO2. The samples were then concentrated using a 0.4 µm polycarbonate track etched, disposable Concentrating Pipette tip. On average, sample process time took 10.1 min (n = 6 ± 1.1), with an average elution volume of 317 µL ± 19.3, with nominal concentration factors of 500×. Moreover, when beer cans were kept at 37°C prior to pouring and incubation at 4°C, the average sample process time averaged 6.5 min. Overall, this demonstrates that American lager can be concentrated to smaller volumes that are more appropriate for rapid detection methods in a relatively short amount of time.
Michael Hornback is director of laboratory operations at InnovaPrep LLC. Michael graduated with his Ph.D. degree in microbiology from East Carolina University in 2006 and has held post-doctoral fellowship positions at Emory University and Kansas University Medical Center. He has over 10 years of experience in the area of bacterial research, with a heavy emphasis on molecular biological techniques. Michael is a senior scientist at InnovaPrep and has developed methods for sample processing for the Concentrating Pipette and has established viral, DNA, and protein protocols used in ongoing research and development projects. He is currently director of laboratory operations and supervises InnovaPrep R&D and demonstration testing.
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