Ultrasonically enhanced fractionation of milk fat in a litre-scale prototype vessel

The ultrasonic fractionation of milk fat in whole milk to fractions with distinct particle size distributions was demonstrated using a stage-based ultrasound-enhanced gravity separation protocol. Firstly, a single stage ultrasound gravity separation was characterised after various sonication durations (5–20 min) with a mass balance, where defined volume partitions were removed across the height of the separation vessel to determine the fat content and size distribution of fat droplets. Subsequent trials using ultrasound-enhanced gravity separation were carried out in three consecutive stages. Each stage consisted of 5 min sonication, with single and dual transducer configurations at 1 MHz and 2 MHz, followed by aliquot collection for particle size characterisation of the formed layers located at the bottom and top of the vessel. After each sonication stage, gentle removal of the separated fat layer located at the top was performed.Results indicated that ultrasound promoted the formation of a gradient of vertically increasing fat concentration and particle size across the height of the separation vessel, which became more pronounced with extended sonication time. Ultrasound-enhanced fractionation provided fat enriched fractions located at the top of the vessel of up to 13 ± 1% (w/v) with larger globules present in the particle size distributions. In contrast, semi-skim milk fractions located at the bottom of the vessel as low as 1.2 ± 0.01% (w/v) could be produced, containing proportionally smaller sized fat globules. Particle size differentiation was enhanced at higher ultrasound energy input (up to 347 W/L). In particular, dual transducer after three-stage operation at maximum energy input provided highest mean particle size differentiation with up to 0.9 μm reduction in the semi-skim fractions. Higher frequency ultrasound at 2 MHz was more effective in manipulating smaller sized fat globules retained in the later stages of skimming than 1 MHz. While 2 MHz ultrasound removed 59 ± 2% of the fat contained in the initial sample, only 47 ± 2% was removed with 1 MHz after 3 ultrasound-assisted fractionation stages.     

Temperature effects on the ultrasonic separation of fat from natural whole milk

This study showed that temperature influences the rate of separation of fat from natural whole milk during application of ultrasonic standing waves. In this study, natural whole milk was sonicated at 600 kHz (583 W/L) or 1 MHz (311 W/L) with a starting bulk temperature of 5, 25, or 40 °C. Comparisons on separation efficiency were performed with and without sonication. Sonication using 1 MHz for 5 min at 25 °C was shown to be more effective for fat separation than the other conditions tested with and without ultrasound, resulting in a relative change from 3.5 ± 0.06% (w/v) fat initially, of −52.3 ± 2.3% (reduction to 1.6 ± 0.07% (w/v) fat) in the skimmed milk layer and 184.8 ± 33.2% (increase to 9.9 ± 1.0% (w/v) fat) in the top layer, at an average skimming rate of ∼5 g fat/min. A shift in the volume weighted mean diameter (D[4,3]) of the milk samples obtained from the top and bottom of between 8% and 10% relative to an initial sample D[4,3] value of 4.5 ± 0.06 μm was also achieved under these conditions. In general, faster fat separation was seen in natural milk when natural creaming occurred at room temperature and this separation trend was enhanced after the application of high frequency ultrasound.     

Creaming enhancement in a liter scale ultrasonic reactor at selected transducer configurations and frequencies

Recent research has shown that high frequency ultrasound (0.4–3 MHz), can enhance milkfat separation in small scale systems able to treat only a few milliliters of sample. In this work, the effect of ultrasonic standing waves on milkfat creaming was studied in a 6 L reactor and the influence of different frequencies and transducer configurations in direct contact with the fluid was investigated. A recombined coarse milk emulsion with fat globules stained with oil-red-O dye was selected for the separation trials. Runs were performed with one or two transducers placed in vertical (parallel or perpendicular) and horizontal positions (at the reactor base) at 0.4, 1 and/or 2 MHz (specific energy 8.5 ± 0.6 kJ/kg per transducer). Creaming behavior was assessed by measuring the thickness of the separated cream layer. Other methods supporting this assessment included the measurement of fat content, backscattering, particle size distribution, and microscopy of samples taken at the bottom and top of the reactor. Most efficient creaming was found after treatment at 0.4 MHz in single and double vertical transducer configurations. Among these configurations, a higher separation rate was obtained when sonicating at 0.4 MHz in a vertical perpendicular double transducer setup. The horizontal transducer configuration promoted creaming at 2 MHz only. Fat globule size increase was observed when creaming occurred. This research highlights the potential for enhanced separation of milkfat in larger scale systems from selected transducer configurations in contact with a dairy emulsion, or emulsion splitting in general.     

Ultrasound pressure distributions generated by high frequency transducers in large reactors

The performance of an ultrasound reactor chamber relies on the sound pressure level achieved throughout the system. The active volume of a high frequency ultrasound chamber can be determined by the sound pressure penetration and distribution provided by the transducers. This work evaluated the sound pressure levels and uniformity achieved in water by selected commercial scale high frequency plate transducers without and with reflector plates. Sound pressure produced by ultrasonic plate transducers vertically operating at frequencies of 400 kHz (120 W) and 2 MHz (128 W) was characterized with hydrophones in a 2 m long chamber and their effective operating distance across the chamber’s vertical cross section was determined. The 2 MHz transducer produced the highest pressure amplitude near the transducer surface, with a sharp decline of approximately 40% of the sound pressure occurring in the range between 55 and 155 mm from the transducer. The placement of a reflector plate 500 mm from the surface of the transducer was shown to improve the sound pressure uniformity of 2 MHz ultrasound. Ultrasound at 400 kHz was found to penetrate the fluid up to 2 m without significant losses. Furthermore, 400 kHz ultrasound generated a more uniform sound pressure distribution regardless of the presence or absence of a reflector plate. The choice of the transducer distance to the opposite reactor wall therefore depends on the transducer plate frequency selected. Based on pressure measurements in water, large scale 400 kHz reactor designs can consider larger transducer distance to opposite wall and larger active cross-section, and therefore can reach higher volumes than when using 2 MHz transducer plates.     

Production of particulates from transducer erosion: Implications on food safety

The formation of metallic particulates from erosion was investigated by running a series of transducers at various frequencies in water. Two low frequency transducer sonotrodes were run for 7.5 h at 18 kHz and 20 kHz. Three high frequency plates operating at megasonic frequencies of 0.4 MHz, 1 MHz, and 2 MHz were run over a 7 days period. Electrical conductivity and pH of the solution were measured before and after each run. A portion of the non-sonicated and treated water was partially evaporated to achieve an 80-fold concentration of particles and then sieved through nano-filters of 0.1 μm, 0.05 μm, and 0.01 μm. An aliquot of the evaporated liquid was also completely dried on strips of carbon tape to determine the presence of finer particles post sieving. An aliquot was analyzed for detection of 11 trace elements by Inductively Coupled Plasma Mass Spectroscopy (ICPMS). The filters and carbon tapes were analyzed by FE-SEM imaging to track the presence of metals by EDS (Energy Dispersive Spectroscopy) and measure the particle size and approximate composition of individual particles detected. Light microscopy visualization was used to calculate the area occupied by the particles present in each filter and high resolution photography was used for visualization of sonotrode surfaces. The roughness of all transducers before and after sonication was tested through profilometry. No evidence of formation of nano-particles was found at any tested frequency. High amounts of metallic micron-sized particles at 18 kHz and 20 kHz formed within a day, while after 7 day runs only a few metallic micro particles were detected above 0.4 MHz. Erosion was corroborated by an increase in roughness in the 20 kHz tip after ultrasound. The elemental analysis showed that metal leach occurred but values remained below accepted drinking water limits, even after excessively long exposure to ultrasound. With the proviso that the particles measured here were only characterized in two dimensions and could be nanoparticulate in terms of the third dimension, this research suggests that there are no serious health implications resulting from the formation of nanoparticles under the evaluation conditions. Therefore, high frequency transducer plates can be safely operated in direct contact with foods. However, due to significant production of metallic micro-particulates, redesign of lower frequency sonotrodes and reaction chambers is advised to enable operation in various food processing direct-contact applications.     

Cavitation and non-cavitation regime for large-scale ultrasonic standing wave particle separation systems – In situ gentle cavitation threshold determination and free radical related oxidation

We here suggest a novel and straightforward approach for liter-scale ultrasound particle manipulation standing wave systems to guide system design in terms of frequency and acoustic power for operating in either cavitation or non-cavitation regimes for ultrasound standing wave systems, using the sonochemiluminescent chemical luminol. We show that this method offers a simple way of in situ determination of the cavitation threshold for selected separation vessel geometry. Since the pressure field is system specific the cavitation threshold is system specific (for the threshold parameter range). In this study we discuss cavitation effects and also measure one implication of cavitation for the application of milk fat separation, the degree of milk fat lipid oxidation by headspace volatile measurements. For the evaluated vessel, 2 MHz as opposed to 1 MHz operation enabled operation in non-cavitation or low cavitation conditions as measured by the luminol intensity threshold method. In all cases the lipid oxidation derived volatiles were below the human sensory detection level. Ultrasound treatment did not significantly influence the oxidative changes in milk for either 1 MHz (dose of 46 kJ/L and 464 kJ/L) or 2 MHz (dose of 37 kJ/L and 373 kJ/L) operation.     

Lipid oxidation volatiles absent in milk after selected ultrasound processing

Ultrasonic processing can suit a number of potential applications in the dairy industry. However, the impact of ultrasound treatment on milk stability during storage has not been fully explored under wider ranges of frequencies, specific energies and temperature applications. The effect of ultrasonication on lipid oxidation was investigated in various types of milk. Four batches of raw milk (up to 2 L) were sonicated at various frequencies (20, 400, 1000, 1600 and 2000 kHz), using different temperatures (4, 20, 45 and 63 °C), sonication times and ultrasound energy inputs up to 409 kJ/kg. Pasteurized skim milk was also sonicated at low and high frequency for comparison. In selected experiments, non-sonicated and sonicated samples were stored at 4 °C and were drawn periodically up to 14 days for SPME–GCMS analysis. The cavitational yield, characterized in all systems in water, was highest between 400 kHz and 1000 kHz. Volatile compounds from milk lipid oxidation were detected and exceeded their odor threshold values at 400 kHz and 1000 kHz at specific energies greater than 271 kJ/kg in raw milk. However, no oxidative volatile compounds were detected below 230 kJ/kg in batch systems at the tested frequencies under refrigerated conditions. Skim milk showed a lower energy threshold for oxidative volatile formation. The same oxidative volatiles were detected after various passes of milk through a 0.3 L flow cell enclosing a 20 kHz horn and operating above 90 kJ/kg. This study showed that lipid oxidation in milk can be controlled by decreasing the sonication time and the temperature in the system depending on the fat content in the sample among other factors.     

Advances in high frequency ultrasound separation of particulates from biomass

In recent years the use of high frequency ultrasound standing waves (megasonics) for droplet or cell separation from biomass has emerged beyond the microfluidics scale into the litre to industrial scale applications. The principle for this separation technology relies on the differential positioning of individual droplets or particles across an ultrasonic standing wave field within the reactor and subsequent biomass material predisposition for separation via rapid droplet agglomeration or coalescence into larger entities. Large scale transducers have been characterised with sonochemiluminescence and hydrophones to enable better reactor designs. High frequency enhanced separation technology has been demonstrated at industrial scale for oil recovery in the palm oil industry and at litre scale to assist olive oil, coconut oil and milk fat separation. Other applications include algal cell dewatering and milk fat globule fractionation. Frequency selection depends on the material properties and structure in the biomass mixture. Higher frequencies (1 and 2 MHz) have proven preferable for better separation of materials with smaller sized droplets such as milk fat globules. For palm oil and olive oil, separation has been demonstrated within the 400–600 kHz region, which has high radical production, without detectable impact on product quality.     

Design parameters of stainless steel plates for maximizing high frequency ultrasound wave transmission

This work validated, in a higher frequency range, the theoretical predictions made by Boyle around 1930, which state that the optimal transmission of sound pressure through a metal plate occurs when the plate thickness equals a multiple of half the wavelength of the sound wave. Several reactor design parameters influencing the transmission of high frequency ultrasonic waves through a stainless steel plate were examined. The transmission properties of steel plates of various thicknesses (1–7 mm) were studied for frequencies ranging from 400 kHz to 2 MHz and at different distances between plates and transducers. It was shown that transmission of sound pressure through a steel plate showed high dependence of the thickness of the plate to the frequency of the sound wave (thickness ratio). Maximum sound pressure transmission of ∼60% of the incident pressure was observed when the ratio of the plate thickness to the applied frequency was a multiple of a half wavelength (2 MHz, 6 mm stainless steel plate). In contrast, minimal sound pressure transmission (∼10–20%) was measured for thickness ratios that were not a multiple of a half wavelength. Furthermore, the attenuation of the sound pressure in the transmission region was also investigated. As expected, it was confirmed that higher frequencies have more pronounced sound pressure attenuation than lower frequencies. The spatial distribution of the sound pressure transmitted through the plate characterized by sonochemiluminescence measurements using luminol emission, supports the validity of the pressure measurements in this study.     

Ultrasonic imaging using air-coupled P(VDF/TrFE) transducers at 2 MHz

A reflection non-contact ultrasonic microscope system working both in amplitude and phase difference modes at 2 MHz has been developed using an air-coupled concave transducer made of piezoelectric polymer films of poly(vinylidene fluoride/trifluoroethylene) [P(VDF/TrFE)]. The transducer is composed of three 95 μm-thick P(VDF/TrFE) films stacked together, each of which is activated electrically in parallel by a driving source. The transducer has a wide aperture angle of 140° and a focal length of 10 mm. The measured two-way transducer insertion loss is 80 dB at 1.83 MHz. Despite 20 dB higher insertion loss than that estimated from Mason’s equivalent circuit, we have obtained clear amplitude acoustic images of a coin with transverse resolution of 150 μm, and clear phase difference acoustic images of the rough surface of a paper currency bill with depth resolution of sub-micrometer. Using two planar transducers of P(VDF/TrFE), we have also successfully measured in through-transmission mode the sound velocity and absorption of a 3 mm-thick silicone-rubber plate. The present study proves that, owing to its low acoustic impedance and flexibility, P(VDF/TrFE) piezoelectric film is very useful for high frequency acoustic imaging in air in the MHz range.     

Fabrication of broadband poly(vinylidene difluoride-trifluroethylene) line-focus ultrasonic transducers for surface acoustic wave measurements of anisotropy of a (1 0 0) silicon wafer

This paper investigates a new method for fabrication of broadband line-focus ultrasonic transducers by sol–gel spin-coating the poly(vinylidene difluoride-trifluroethylene) [P(VDF-TrFE)] copolymer film on a concave fine-polished beryllium copper backing. The ferroelectric hysteresis loops of the P(VDF-TrFE) films spin-coated from different molar ratios of VDF/TrFE, 77/23 and 55/45, were measured to select the better mixture. Owing to the better acoustic matching to water, compared with lead zirconate titanate (PZT), the fabricated transducers show relatively wide bandwidth of approximately 50 MHz with high central frequency of 60 MHz obtained at the focal plane when a fused-quartz acts as a reflecting target. Each one of the two finished transducers has a focal length of 5 mm and a full aperture angle of 90°. After applying the specially developed digital signal processing algorithm to the defocusing experiment data, which is called V(f,z) analysis method based on two-dimensional fast Fourier transform (2-D FFT), the operating frequency can extend from several MHz to over 90 MHz. Surface acoustic wave (SAW) velocities of a typical (1 0 0) silicon wafer was measured along various directions between [1 0 0] and [0 1 0] to represent the anisotropic features.     

Efficient high voltage pulser for piezoelectric air coupled transducer

The design of high voltage pulser for air coupled ultrasound imaging is presented. It is dedicated for air-coupled ultrasound applications when piezoelectric transducer design is used. Two identical N-channel MOSFETs are used together with 1200 V high and low side driver IC. Simple driving pulses’ delay and skew circuit is used to reduce the cross-conduction. Analysis of switch peak current and channel resistance relation to maximum operation frequency and load capacitance is given. PSPICE simulation was used to analyze the gate driver resistance, gate pulse skew, pulse amplitude influence on energy consumption when loaded by capacitive load. Experimental investigation was verified against simulation and theoretical predictions. For 500 pF capacitance, which is most common for piezoelectric air coupled transducers, pulser consumes 650 μJ at 1 kV pulse and 4 μJ at 50 V. Pulser is capable to produce up to 1 MHz pulse trains with positive 50 V–1 kV pulses with up to 10 A peak output current. When loaded by 200 kHz transducer at 1 kV pulse amplitude rise time is 40 ns and fall time is 32 ns which fully satisfies desired 1 MHz bandwidth.     

Ultrasound-assisted transesterification of soybean oil using low power and high frequency and no external heating source

In this work, high frequency and low power ultrasound without external heating source and mechanical stirring in biodiesel production were studied. Transesterification of soybean oil with methanol and catalyzed by KOH was investigated using ultrasound equipment and ultrasonic transducer. The effect of ultrasonic output power (3 W–9 W), ultrasonic frequency (1 MHz and 3 MHz), and alcohol to oil molar ratio (6:1 and 8:1) have been investigated. The increase in ultrasonic power provided higher conversion rates. In addition, higher conversion rates were obtained by increasing the ultrasonic frequency from 1 MHz to 3 MHz (48.7% to 79.5%) for the same reaction time. Results also indicate that the speed of sound can be used to evaluate the produced biodiesel qualitatively. Further, the ultrasound system presented electric consumption (46.2 W∙h) four times lower than achieved using the conventional method (211.7 W∙h and 212.3 W∙h). Thus, biodiesel production using low power ultrasound in the MHz frequency range is a promising technology that could contribute to biodiesel production processes.     

Cavitation mapping by sonochemiluminescence with less bubble displacement induced by acoustic radiation force in a 1.2 MHz HIFU

An acoustic radiation force counterbalanced appliance was employed to map the cavitation distribution in water. The appliance was made up of a focused ultrasound transducer and an aluminum alloy reflector with the exactly same shape. They were centrosymmetry around the focus of the source transducer. Spatial–temporal dynamics of cavitation bubble clouds in the 1.2 MHz ultrasonic field within this appliance were observed in water. And they were mapped by sonochemiluminescence (SCL) recordings and high-speed photography. There were significant differences in spatial distribution and temporal evolution between normal group and counterbalanced group. The reflector could avoid bubble directional displacement induced by acoustic radiation force under certain electric power (⩽50 W). As a result, the SCL intensity in the pre-focal region was larger than that of normal group. In event of high electric power (⩾70 W), most of the bubbles were moving in acoustic streaming. When electric power decreased, bubbles kept stable and showed stripe structure in SCL images. Both stationary bubbles and moving bubbles have been captured, and exhibited analytical potential with respect to bubbles in therapeutic ultrasound.     

Enhanced OH radical generation by dual-frequency ultrasound with TiO2 nanoparticles: Its application to targeted sonodynamic therapy

The present study demonstrated the enhanced hydroxyl (OH) radical generation by combined use of dual-frequency (0.5 MHz and 1 MHz) ultrasound (US) and titanium dioxide (TiO₂) nanoparticles (NPs) as sonocatalyst. The OH radical generation became the maximum, when 0.5 MHz US was irradiated at an intensity of 0.8 W/cm² and 1 MHz US was irradiated at intensities at 0.4 W/cm² in the presence of TiO₂ NPs under the examined conditions. After incorporation of TiO₂ NPs modified with targeting protein pre-S1/S2, HepG2 cancer cells were subjected to the dual-frequency US at optimum irradiation intensities (“targeted-TiO₂/dual-US treatment”). Growth of the HepG2 cells was reduced by 46% compared with the control condition after irradiation of dual-frequency US for 60 s with TiO₂ NPs incorporation. In contrast, HepG2 cell growth was almost the same as that in the control condition when cells were irradiated with either 0.5 MHz or 1 MHz ultrasound alone without TiO₂ NP incorporation.     

Inactivation of Enterobacter aerogenes in reconstituted skim milk by high- and low-frequency ultrasound

The inactivation of Enterobacter aerogenes in skim milk using low-frequency (20 kHz) and high-frequency (850 kHz) ultrasonication was investigated. It was found that low-frequency acoustic cavitation resulted in lethal damage to E. aerogenes. The bacteria were more sensitive to ultrasound in water than in reconstituted skim milk having different protein concentrations. However, high-frequency ultrasound was not able to inactivate E. aerogenes in milk even when powers as high as 50 W for 60 min were used. This study also showed that high-frequency ultrasonication had no influence on the viscosity and particle size of skim milk, whereas low-frequency ultrasonication resulted in the decrease in viscosity and particle size of milk. The decrease in particle size is believed to be due to the breakup of the fat globules, and possibly to the cleavage of the κ-casein present at the surface of the casein micelles. Whey proteins were also found to be slightly affected by low-frequency ultrasound, with the amounts of α-lactalbumin and β-lactoglobulin slightly decreasing.     

Ultrasonic biodiesel synthesis from crude Jatropha curcas oil with heterogeneous base catalyst: Mechanistic insight and statistical optimization

This paper reports studies in ultrasound-assisted heterogeneous solid catalyzed (CaO) synthesis of biodiesel from crude Jatropha curcas oil. The synthesis has been carried out in two stages, viz. esterification and trans-esterification. The esterification process is not influenced by ultrasound. The transesterification process, however, shows marked enhancement with ultrasound. A statistical experimental design has been used to optimize the process conditions for the synthesis. XRD analysis confirms formation of Ca(OMe)₂, which is the active catalyst for transesterification reaction. The optimum values of parameters for the highest yield of transesterification have been determined as follows: alcohol to oil molar ratio ≈ 11, catalyst concentration ≈ 5.5 wt.%, and temperature ≈ 64 °C. The activation energy of the reaction is calculated as 133.5 kJ/mol. The heterogeneity of the system increases mass transfer constraints resulting in approx. 4× increase in activation energy as compared to homogeneous alkali catalyzed system. It is also revealed that intense micro-convection induced by ultrasound enhances the mass transfer characteristics of the system with ∼20% reduction in activation energy, as compared to mechanically agitated systems. Influence of catalyst concentration and alcohol to oil molar ratio on the transesterification yield is inter-linked through formation of methoxy ions and their diffusion to the oil–alcohol interface, which in turn is determined by the volume fractions of the two phases in the reaction mixture. As a result, the highest transesterification yield is obtained at the moderate values of catalyst concentration and alcohol to oil molar ratio.     

Ultrasound-mediated drug delivery using liposomes modified with a thermosensitive polymer

Ultrasound-mediated drug delivery was established using liposomes that were modified with the thermosensitive polymer (TSP) poly(NIPMAM-co-NIPAM), which sensitized the liposomes to high temperatures. TSP-modified liposomes (TSP liposomes) released encapsulated calcein under 1 MHz ultrasound irradiation at 0.5 W/cm² for 120 s as well as the case under incubation at 42 °C for 15 min. In addition, uptake of the drug released from TSP liposomes by cancer cells was enhanced by ultrasound irradiation. In a cell injury assay using doxorubicin (DOX)-loaded TSP liposomes and ultrasound irradiation, cell viability of HepG2 cells at 6 h after ultrasound irradiation (1 MHz, 0.5 W/cm² for 30 s) with DOX-loaded TSP liposomes (TSP/lipid ratio = 1) was 60%, which was significantly lower than that of the control conditions such as DOX-loaded TSP liposomes alone and DOX-loaded intact liposomes under ultrasound irradiation.     

Lysis of Chlamydomonas reinhardtii by high-intensity focused ultrasound as a function of exposure time

Efficient lysis of microalgae for lipid extraction is an important concern when processing biofuels. Historically, ultrasound frequencies in the range of 10–40 kHz have been utilized for this task. However, greater efficiencies might be achievable if higher frequencies could be used. In our study, we evaluated the potential of using 1.1 MHz ultrasound to lyse microalgae for biofuel production while using Chlamydomonas reinhardtii as a model organism. The ultrasound was generated using a spherically focused transducer with a focal length of 6.34 cm and an active diameter of 6.36 cm driven by 20 cycle sine-wave tone bursts at a pulse repetition frequency of 2 kHz (3.6% duty cycle). The time-average acoustic power output was 26.2 W while the spatial-peak-pulse-average intensity (I_SPPA) for each tone burst was 41 kW/cm². The peak compressional and rarefactional pressures at the focus were 102 and 17 MPa, respectively. The exposure time was varied for the different cases in the experiments from 5 s to 9 min and cell lysis was assessed by quantifying the percentage of protein and chlorophyll release into the supernate as well as the lipid extractability. Free radical generation and lipid oxidation for the different ultrasound exposures were also determined. We found that there was a statistically significant increase in lipid extractability for all of the exposures compared to the control. The longer exposures also completely fragmented the cells releasing almost all of the protein and chlorophyll into the supernate. The cavitation activity did not significantly increase lipid oxidation while there was a minor trend of increased free radical production with increased ultrasound exposure.     

Ultrasound improves the renneting properties of milk

The effects of ultrasound application on skim milk (10% w/w total solids at natural pH 6.7 or alkali-adjusted to pH 8.0) prior to the renneting of milk at pH 6.7 were examined. Skim milk, made by reconstituting skim milk powder, was sonicated at 20 kHz and 30 °C (dissipated power density 286 kJ kg⁻¹) in an ultrasonic reactor. The rennet gelation time, curd firming rate, curd firmness, and the connectivity of the rennet gel network were improved significantly in rennet gels made from milk ultrasonicated at pH 8.0 and re-adjusted back to pH 6.7 compared to those made from milk sonicated at pH 6.7. These renneting properties were also improved in milk sonicated at pH 6.7 compared to those of the non-sonicated control milk. The improvements in renneting behavior were related to ultrasound-induced changes to the proteins in the milk. This study showed that ultrasonication has potential to be used as an intervention to manipulate the renneting properties of milk for more efficient manufacturing of cheese.