This study was performed to investigate the effects of high pressure/high temperature (HPHT) treatment on the recovery efficiency and characteristics of porcine placenta hydrolysates. The placenta hydrolysates were characterized by solubility, free amino acid contents, gel electrophoresis, gel permeation chromatography (GPC) and amino acid composition. Placenta was treated at 37.5 MPa of pressure combined with various temperatures (150, 170, and 200 o C) or various holding times (0, 30, and 60 min at 170 o C). Insoluble raw placenta collagen was partially solubilized (> 60% solubility) by the HPHT treatment. Free amino group content of placenta collagen was increased from 0.1 mM/g collagen to > 0.3 mM/g collagen after HPHT treatment, reflecting partial hydrolysis of collagen. The molecular weight (M w ) distribution showed evidence of collagen hydrolysis by shifting of M w peaks toward low molecular weight when treated temperature or holding time was increased. Alanine (Ala), glycine (Gly), hydroxyproline (Hyp), and proline (Pro) contents increased after the HPHT treatments compared to a decrease in the others. In particular, the increase in Gly was obvious, followed by Hyp and Pro, reflecting that placenta hydrolysates were mainly composed of these amino acids. However, increasing temperature or holding time hardly affected the amino acid compositions. These results indicate that the HPHT treatment is advantageous to hydrolyze collagen derived from animal by-products.

This study investigated the effects of three proteases (trypsin, pepsin and chymotrypsin) on the hydrolysis efficiency of porcine placenta and the molecular weight (Mw) distributions of the placental hydrolysates. Because placenta was made up of insoluble collagen, the placenta was gelatinized by applying thermal treatment at 90 ℃ for 1 h and used as the sample. The placental hydrolyzing activities of the enzymes at varying concentrations and incubation times were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel permeation chromatography (GPC). Based on the SDS-PAGE, the best placental hydrolysis efficiency was observed in trypsin treatments where all peptide bands disappeared after 1 h of incubation as compared to 6 h of chymotrypsin. Pepsin hardly hydrolyzed the placenta as compared to the other two enzymes. The Mw distribution revealed that the trypsin produced placental peptides with Mw of 106 and 500 Da. Peptides produced by chymotrypsin exhibited broad ranges of Mw distribution (1-20 kDa), while the pepsin treatment showed Mw greater than 7 kDa. For comparisons of pre-treatments, the subcritical water processing (37.5 MPa and 200 ℃ of raw placenta improved the efficiency of tryptic digestions to a greater level than that of a preheating treatment (90 ℃ for 1 h). Consequently, subcritical water processing followed by enzymatic digestions has the potential of an advanced collagen hydrolysis technique.

In a previous study, hydrolysates of porcine placenta were obtained and the extraction efficiency for proteins and amino acids was compared between sub- and super-critical water extraction systems; optimum efficiency was found to be achieved using subcritical water (170℃, 10 bar). In this study, the effects of adding ethanol to the subcritical water system were investigated. The lowest-molecular-weight extraction product detected weighed 434 Da, and the efficiency of extraction for low-molecular-weight products was increased when either the concentration of ethanol was decreased, or the extraction time was lengthened from 10 min to 30 min. The highest concentration of free amino acids (approximately 8 mM) was observed following 30 min extraction using pure distilled water. The concentration of free amino acids was significantly lower when ethanol was added or a shorter extraction time was used (p<0.05). Color change of the solution following extraction was measured. There were no significant differences in color between lysates produced with different extraction times when using distilled water (p>0.05); however, using different extraction times produced significant differences in color when using 20% or 50% ethanol solution for subcritical extraction (p<0.05). The range of pH for the hydrolysate solutions was 6.4-7.5. In conclusion, the investigated extraction system was successful in the extraction of ≤ 500 Da hydrolysates from porcine placenta, but addition of ethanol did not yield higher production of low-molecular-weight hydrolysates than that achieved by DW alone.

This study was carried out to investigate the effect of trans-cinnamaldehyde and high pressure treatment on milk. Cinnamon oil milk was manufactured by high speed homogenization (3,000 rpm) and high pressure homogenization (500 and 2,000 bar) processing UHT milk and trans-cinnamaldehyde of various concentrations (0 to 0.1% (w/v)). Cinnamon oil milk was inoculated with Escherichia coli (6.4 Log CFU/mL) and kept at 7 o C for 10 d to observe the antibacterial effect. The cinnamon oil milk containing 0.05% (w/v) trans-cinnamaldehyde initially began to show an antibacterial effect and Escherichia coli completely died in cinnamon oil milk added 0.1% (w/v) trans-cinnamaldehyde on the 6th day of storage. The result of the TBA value showed that the addition of 0.1% (w/v) trans-cinnamaldehyde was also effective to protect lipid oxidation. In the physical properties of cinnamon oil milk, particle sizes were enlarged in all samples during storage periods and the total color difference of cinnamon oil milk was slightly increased as level of high pressure. The surface tension of cinnamon oil milk treated 2,000 bar was remarkably higher than other samples. It seems that trans-cinnamaldehyde showed antibacterial activity and antioxidation effect at 0.05 and 0.1% (w/v) of concentration. Remarkably, high pressure treatment did not influence its microbial property but slightly affected the physical properties of cinnamon oil milk.

Combined high pressure and subzero temperature phase transition was performed to inactivate Escherichia coli ATCC 10536 at 200 MPa. Inactivation was compared among atmospheric pressure inactivation (APFI), pressure assisted freezing inactivation (PAFI), pressure assisted thawing inactivation (PATI), and pressure shift freezing inactivation (PSFI). Phase transition characteristics of microbial suspension were concurrently analyzed to investigate its influence on microbial inactivation. Phase transition temperature of E. coli ATCC 10536 suspension decreased to -23.8°C at 200 MPa, which is observed from the phase transition plateau. Supercooling was observed from nucleation temperature (-25°C) of PAFI. PATI resulted in the most inactivation of 4.37 log reduction, followed by PAFI and PSFI in decreasing order. APFI showed no efficacy of inactivation for E. coli ATCC 10536. Synergistic effect of high pressure, subzero temperature, and phase transition was expected for microbial inactivation.