A limit of detection of 200 CFU/mL of spiked in various sample matrices were achieved in 30 min. The sample matrices were raw/unprocessed milk, commercially available milk, juice from packed bottles, fresh juice from carts, potable water, turbid water and calf serum. The complete protocol comprised of three steps: (a) cell lysis (b) nucleic acid amplification and (c) an in situ optical detection. The cell lysis was carried out using a simple heating based protocol, while the loop-mediated isothermal amplification of DNA was carried out by an in-house designed and fabricated system. The developed system consists of an aluminum block fitted with two cartridge heaters along with a thermocouple. The system was coupled to a light source and spectrometer for a simultaneous in situ detection. Primers specific for STY2879 gene were used to amplify the nucleic acid sequence, isolated from cells. The protocol involves 15 min of cell lysis and DNA isolation followed by 15 min for isothermal amplification and simultaneous detection. No cross-reactivity of the primers were observed at 10 CFU/mL of ,, ,, . In addition, the system was able to detect of 200 CFU/mL in a concoction of 10 CFU/mL of , 10 CFU/mL of , and 10 CFU/mL of hepatocyte-derived cellular carcinoma HUH7 cells. The proposed rapid diagnostic system shows a promising future in the field of food and medical diagnostics.
The performance of nano-filtration (NF) for separating phenolic compounds from sugar in apple juice was studied using 1 and 0.25 kDa molecular weight cut-off (MWCO) spiral wound membranes. If these phenolic compounds could be recovered, they could stabilize the juice from haze formation or be added as antioxidants to foods and beverages in order to increase their health properties. Batch experiments were conducted on a pilot scale rig using a diluted clear apple juice concentrate. For the 1 kDa MWCO membrane, the research determined the effect of operating conditions on process efficiency and membrane fouling. The concentration of polyphenolics on the retentate side increased by a factor of up to 4 and the sugar concentration increased by 1.5 times under optimum conditions of lower temperature (30oC), acidic pH (2), lower transmembrane pressure (5 Bar) and higher initial sugar concentration (20 oBrix). Despite the increase in polyphenolics in the retentate, there was little difference in the phenolic composition between retentate and permeate solutions. As the molecular mass of the rejected phenolics was smaller than the membrane cut-off, this indicated that the rejection was related to the formation of a secondary membrane formed as a result of fouling. A mass balance of polyphenolics in the final retentate and permeate compared with the initial feed solution indicated that up to 4.3 gm of polyphenolics were bound per m2 of membrane. The permeate solutions collected from the 1 kDa MWCO membrane were then filtered using a 0.25 kDa MWCO membrane. Most phenolic compounds were retained by the membrane and the concentration increased by a factor of up to 2. Catechin, rutin, phloridzin and quercetin derivatives were concentrated on the retentate side. However, around 20 -40% of chlorogenic acid and epicatechin was observed on the permeate side. It is concluded that membrane separation represents a potentially efficient and cost-effective technology to separate the phenolic fraction of fruit juice in a form suitable for use as a functional ingredient.
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