The impact of pulsed electric fields (PEF) on cellular integrity and texture of Ranchero and Sabroso onions (Allium cepa L.) was investigated. Electrical properties, ion leakage rate, texture, and amount of enzymatically formed pyruvate were measured before and after PEF treatment for a range of applied field strengths and number of pulses. Critical electric field strengths or thresholds (E(c)) necessary to initiate membrane rupture were different because dissimilar properties were measured. Measurement of electrical characteristics was the most sensitive method and was used to detect the early stage of plasma membrane breakdown, while pyruvate formation by the enzyme alliinase was used to identify tonoplast membrane breakdown. Our results for 100-μs pulses indicate that breakdown of the plasma membrane occurs above E(c)= 67 V/cm for 10 pulses, but breakdown of the tonoplast membrane is above either E(c)= 200 V/cm for 10 pulses or 133 V/cm for 100 pulses. This disparity in field strength suggests there may be 2 critical electrical field strengths: a lower field strength for plasma membrane breakdown and a higher field strength for tonoplast membrane breakdown. Both critical electric field strengths depended on the number of pulses applied. Application of a single pulse at an electric field up to 333 V/cm had no observable effect on any measured properties, while significant differences were observed for n≥10. The minimum electric field strength required to cause a measurable property change decreased with the number of pulses. The results also suggest that PEF treatment may be more efficient if a higher electric field strength is applied for a fewer pulses.

Pulsed electric fields (PEF) nonthermally induce cell membrane permeabilization and thereby improve dehydration and extraction efficiencies in food plant materials. Effects of electrical field strength and number of pulses on plant tissue integrity have been studied extensively. Two previous studies on the effect of pulse frequency, however, did not provide a clear view: one study suggested no effect of frequency, while the other found a greater impact on tissue integrity at lower frequency. This study establishes the effect of pulse frequency on integrity of onion tissues. Changes in electrical characteristics, ion leakage, texture parameters, and percent weight loss were quantified for a wide range of pulse frequencies under conditions of fixed field strength and pulse number. Optical microscopy and viable-cell staining provided direct visualization of effects on individual cells. The key finding is that lower frequencies (f < 1 Hz) cause more damage to tissue integrity than higher frequencies (f = 1 to 5000 Hz). Intriguingly, the optical microscopy observations demonstrate that the speed of intracellular convective motion (that is, cytoplasmic streaming) following PEF application is strongly correlated with PEF frequency. We provide the first in situ visualization of the intracellular consequence of PEF at different frequencies in a plant tissue. We hypothesize that cytoplasmic streaming plays a significant role in moving conductive ionic species from permeabilized cells to the intercellular space between plant cells, making subsequent pulses more efficacious at sufficiently low frequencies. The results suggest that decreasing the pulse frequency in PEF may minimize the number of pulses needed to achieve a desired amount of permeabilization, thus lowering the total energy consumption. Practical Application: PEF cause pores to be formed in plant cell membranes, thereby improve moisture removal and potential extraction of desirable components. This study used in situ microscopic evaluation of onion cells, as they were pulsed with electric fields at different frequencies, to determine whether frequency was an important parameter. We illustrate that membranes were more effectively broken at lower frequencies as compared to higher frequencies. Application of this information will allow for improved design of PEF systems for more energy efficient dehydration or extraction of plant tissues.

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Most of the food processing operations involve the use of heat which generally causes alteration, and degradation of natural pigments, resulting in lower stability. One of the stability enhancement methods is co-pigmentation. This study aimed to determine effect of catechin co-pigment on stability of anthocyanins in Clitoria ternatea (or butterfly pea flower) extract. Degradation kinetics of anthocyanins in the extract were evaluated at three temperatures (28, 60, and 90℃). The effect of co-pigment ratio (catechin: anthocyanins at 1:1, 50:1 and 100:1 by weight) on the stability of anthocyanin extract at 90℃ was determined by the pH differential method. It was found that anthocyanin degradation followed the zero- order kinetics at all temperatures; the degradation rate increased as the temperature increased. At a lower pH, anthocyanins became more stable. An increase in the co-pigment ratio significantly retarded the degradation anthocyanins at 90℃. In addition, co-pigmentation also intensified the color of butterfly pea extract. The highest anthocyanin stability was obtained at co-pigment ratio of 100:1.

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Pulsed electric fields (PEF) are known to be effective at permeabilizing plant tissues. Prior research has demonstrated that lower pulse frequencies induce higher rates of permeabilization, but the underlying reason for this response is unclear. Intriguingly, recent microscopic observations with onion tissues have also revealed a correlation between PEF frequency and the subsequent speed of intracellular convective motion, i.e., cytoplasmic streaming. In this paper, we investigate the effect of cytoplasmic streaming on the efficacy of plant tissue permeabilization via PEF. Onion tissue samples were treated with Cytochalasin B, a known inhibitor of cytoplasmic streaming, and changes in cellular integrity and viability were measured over a wide range of frequencies and field strengths. We find that at low frequencies (f < 1 Hz), the absence of cytoplasmic streaming results in a 19% decrease in the conductivity disintegration index compared with control samples. Qualitatively, similar results were observed using a microscopic cell viability assay. The results suggest that at low frequencies convection plays a statistically significant role in distributing more conductive fluid throughout the tissue, making subsequent pulses more efficacious. The key practical implication is that PEF pretreatment at low frequency can increase the rate of tissue permeabilization in dehydration or extraction processes, and that the treatment will be most effective when cytoplasmic streaming is most active, i.e., with freshly prepared plant tissues.

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