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Effect of Thawing and Cold Storage on Frozen Chicken Thigh Meat Quality by High-Voltage Electrostatic Field

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ABSTRACT:  One of the most popular issues in electrostatic biology is the effects of a high-voltage electrostatic field (HVEF) on the thawing of chicken thigh meat. In this study, chicken thigh meat was treated with HVEF (E-group), and compared to
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         M      :       F      o      o       d       M       i      c      r      o       b       i      o       l      o      g      y       &       S      a       f      e      t      y  JFS  M: Food Microbiology and Safety EffectofThawingandColdStorageonFrozenChickenThighMeatQualitybyHigh-VoltageElectrostaticField C HANG -W  EI H SIEH ,C HENG -H UNG L  AI ,W   AI -J  ANE H O ,S U -C HEN H UANG ,  AND  W  EN -C HING K  O  ABSTRACT: Oneofthemostpopularissuesinelectrostaticbiologyistheeffectsofahigh-voltageelectrostaticfield(HVEF) on the thawing of chicken thigh meat. In this study, chicken thigh meat was treated with HVEF (E-group),andcomparedtosamplesstoredinacommonrefrigerator(R-group),toinvestigatehowHVEFaffectschickenthighmeat quality after thawing at low temperature storage ( − 3 and 4  ◦ C). The results showed that there were no signif-icant differences in biochemical and microorganism indices at − 3  ◦ C. However, the HVEF can significantly shortenthawing time for frozen chicken thigh meat at − 3  ◦ C. After thawing chicken thigh meat and storing at 4  ◦ C, the to-tal viable counts reached the Intl. Commission on Microbiological Specification for Foods limit of 10 7 CFU/g onthe 6 and 8 d for the R- and E-group, respectively. On the 8th d, the volatile basic nitrogen had increased from11.24 mg/100 g to 21.9 mg/100 g for the E-group and 39.9 mg/100 g for the R-group, respectively. The biochemi-calandmicroorganismindicesalsoindicatedthattheE-grouptreatmentyieldedbetterresultsonthawingthantheR-grouptreatment.Theapplicationofthismodelhasthepotentialtokeepproductsfresh.Keywords: coldstorage,freshnesskeeping,frozenchickenmeat,high-voltageelectrostaticfield(HVEF),thawing  Introduction T he methodology and techniques used in freezing and thaw-ing processes play an important role in the preservation of the quality of frozen foods. Freezing and thawing rates have beenknown to affect internal meat structure to varying degrees throughcellular disruption by ice crystal formation (Mandigo and Osburn1996). Both the importance of freezing meats and the correct pro-cedures for freezing have been extensively studied (Kinsman andothers 1994). However, the process of thawing has been largely ig-nored. Thawing involves risks such as meat discoloration, weightdecrease (such as thawing loss, cooking loss), and oxidationof lipids due to prolonged thawing time (Kinsman and others1994; Foegeding and others 1996; otto and others 2004). More-over, during thawing, foods are subject to damage by chemicaland physical changes and the action of microorganisms. There-fore, optimum thawing procedures should be of concern to foodtechnologists (Kalichevsky and others 1995).Current methods used for food thawing are typically slow oruneven resulting in a decrease in food quality (Anderson andSingh 2006). To avoid the deleterious effects of heat on the flavor,color, and nutrient value of the food, nonthermal technology areused for food thawing (Barbosa-Canovas and others 1998). Electricfields, magnetic fields, high hydrostatic pressure (HHP) and ultra-high pressure are some of nonthermal technologies (Shazmanaand others 2007). As the electrostatic field can affect the perme-ability of cell membranes and enzymes activity (Dimitrov 1984;Barsotti and Cheftel 1999), the application of high-voltage electro- MS 20090633 Submitted 7/2/2009, Accepted 2/4/2010. Authors Hsieh and Ho are with Dept. of Medicinal Botanicals and Health Care, Da-Yeh Univ.,168 Univ. Rd, Da-Tsuen, Chang-Hua, Taiwan, Republic of China. Authors Lai, Huang, and Ko are with Dept. and Graduate Program of BioIndustry Technology, Da-Yeh Univ., 168 Univ. Rd, Da-Tsuen, Chang-Hua, Taiwan,Republic of China. Direct inquiries to author Ko (E-mail: welson6404@ hotmail.com). static field (HVEF) in maintenance of food freshness has becomeone of the most promising technologies. According to the previousstudies, the negative electrons of HVEF have effects on cell mem-brane (Dimitrov 1984). Zhao and others (2007) have reported thatthe exceeded applied electrostatic field may cause an alteration inprotein secondary structure. There are many HVEF applicationssuch as suppressing the respiration rate, extended freshness, andimprovingproductshelflife(forexample,sweetpeppersandstraw-berries) (Kharel and others 1996), as well as in accelerating theinhibition of microbial growth (Kharel and others 1996; Cao andothers 2004). However, a refrigerating system based on HVEF forlow-temperature thawing has never been reported.The chicken thigh meat was initially frozen at approximately  − 1 ◦ C.Only 2/3of the water was frozen at − 3 ◦ C.One of theindicestested was the time taken to reach  − 5  ◦ C, the lower temperatureof the zone of maximum ice crystal formation. This study investi-gatedtheeffectofthethawingandstorageprocessonthefreshnessquality of chicken thigh meat using HVEF at − 3 and 4  ◦ C. Materials and Methods Materials The fresh chicken thigh meat were purchased from a local retailmarket,killedimmediately,andkeptonicebeforesampleprepara-tion. All chemicals used in this study were purchased from Sigma- Aldrich-Fluka Chemical Co. (St. Louis, Mo., U.S.A.). Samplepreparation Skinless and boneless chicken thigh meat, cut into pieces of ap-proximately 190 g and 2.8 cm in thickness by knife after use of thequick freezing method storage and keep at − 21  ◦ C for 48 h. C   2010 Institute of Food Technologists R  Vol. 75, Nr. 4, 2010 —  JOURNAL OF FOOD SCIENCE  M 193 doi: 10.1111/j.1750-3841.2010.01594.x Further reproduction without permission is prohibited  M  :    F     o    o    d    M  i      c   r    o    b    i      o   l      o     g     y    &    S     a   f      e   t       y    Effect of high-voltage electrostatic field on the frozen chicken thigh meat. .. HVEFrefrigeratingsystem The schematic diagram for HVEF refrigerating system (HRS) isshown in Figure 1. It mainly consists of a multiple points-to-plateelectrode system, 2 treatment chambers, an incubator, and a high-voltage power supply (Model Chargemaster CM-60, Taiwan Hon-lycon, Inc., Taiwan). The sharp points of 16 needles (0.001 mm india.), which were connected to the negative pole of a high-voltagepower unit, formed the corona discharge electrode. The voltagecan be adjusted from 0 to 60 kV by a controller. The groundedplane electrode was a 58 × 45 cm rectangle stainless steel plate. A control with a dummy electrode system was not connected to thepower unit. The 2 treatment chambers were placed in the incuba-tor for temperature control. The distance (discharge gap) betweenthe corona electrode and plate electrode was adjustable. The volt-age for the electrode system is 20 kV, the distance between plates is0.2 m, and the electrostatic field strength is 100 kV/m. E   = V D  ,  where  E   is the electrostatic field strength (kV/m),  V   is the outputvoltage (kV), and  D   is the distance between plates (m). Thawingprocess The chicken thigh meat in the E-group was thawed in a − 3 and4  ◦ C refrigerator equipped with a high-electrostatic voltage field.The chicken thigh meat in the R-group was stored in a common re-frigerator at the same temperature. The biochemical, microbiolog-ical, and physical experiments were carried out after 8 d of storage.Thecentertemperaturesofeachsampleweredeterminedbydigitalthermometer (T-60 Digital Thermometer, Type K Thermo Couple,Rixen Co., Ltd., Taiwan). Measurementofthawingloss The samples were placed in polyethylene bags during thawing.Thawinglosswasdeterminedbyweighing6samplespertreatmentbeforemeatthawingandthenreweighedafteraspecifictreatment.Thawing loss parameter was determined by Thawingloss = P  0 − P  F P  0 × 100% ,  where  P  0  is the initial sample weight (before thawing) and  P  F  is thefinal sample weight (after application of a specific treatment). Forfrozen samples, weight was obtained after thawing (Fern´andez andothers 2007). Measurementofcookingloss The thawed meat were placed in polyethylene bags and cookedin a 75  ◦ C water bath for 30 min until an internal temperature of 72  ◦ C was reached. Cooking loss parameter was determined using the relationshipCookingloss = P  0 − P  F P  0 × 100% ,  where  P  0  is the initial sample weight (before cooking) and  P  F  isthe final sample weight (after application of a specific treatment)(Barbantia and Pasquini 2004). Measurementofwaterholdingcapacity   A method proposed by Fern´andez and others (2007) was mod-ified to measure the WHC. Thawed meat weighing 10 g wasintroduced into centrifuge tubes with 2 micropipette filters (What-man nr 4) and was centrifuged at 3000  g   (KUBOTA 6800, KubotaCo., Osaka, Japan) for 20 min at 4  ◦ C. The thawed meat was then Figure 1---Schematic diagram of thehigh-voltage electrostatic field(HVEF) refrigerating system.A. voltage regulator; B. operatefaceplate; C. power switch;D. supporting insulator; E. wafter;F. copper plate; G. sample;H. earthing. M 194  JOURNAL OF FOOD SCIENCE — Vol. 75, Nr. 4, 2010         M      :       F      o      o       d       M       i      c      r      o       b       i      o       l      o      g      y       &       S      a       f      e      t      y Effect of high-voltage electrostatic field on the frozen chicken thigh meat. . . reweighed (once filter paper-dried) and its WHC was determinedusing the relationship WHC =  1 −   P  0 − P  F P  F ×  Watercontent  × 100% ,  where  P  0  is the initial sample weight (before centrifugation) and P  F is the final sample weight (after centrifugation). Six samples wereanalyzed in this study. The measurements were repeated in tripli-cate for each sample. Measurementoftotalviablecounts Samples (10 g for each) were prepared from the thawed meatby homogenizing (Oster 869-18, Mexico) with 90 mL sterilized dis-tilled water. Multiple serial dilutions of the homogenates were pre-pared using sterile normal saline water. Total aerobic bacterialplatecountsweredeterminedusingspreadplatesofStandardPlateCountAgar(PCA)(Difco TM ,BectonDickinsonandCo.,Sparks,Md.,U.S.A.). Each diluent was spread on each agar plate and then incu-bated for 48 h at 37  ◦ C (AOAC 2002). Measurementoftotalvolatilebasicnitrogen Minced thawed meat weighing 10 g was mixed with 90 mL dis-tilled water and then transferred into a distillation flask contain-ing 100 mL distilled water. After the addition of 2 g of magnesiumoxide and an antifoaming agent, the mixture was distilled using themicroKjeldahldistillationapparatus.Distillatewascollectedfor25mininto25mLof4g/100gboricacidand5dropsofTasheroin-dicator. The solution was titrated using (0.1 M) HCl to calculate thetotalvolatilebasicnitrogeninthesampleintermsofmgVBN/100g thawed meat (Pearson 1976). MeasurementofpH The pH value of thawed meat was determined by 6 samples pertreatment and measurements were repeated in triplicate for eachsample. The sample was thoroughly homogenized with 10 mL of distilled water and the homogenate was used for determinationof the pH using pH meter (TOA Electronics Ltd., HM-20S, Japan)(Ngapo and others 1999). Statisticalanalysis Thestatisticalanalysissystemwasadoptedtoperformdataanal- ysis and statistical computations for analysis of variance (ANOVA)and Duncan’s test. Significance of differences was defined at  P   < 0.05. The differences among treatments were verified by their leastsignificant difference. Results and Discussion Thawingcurveoffrozenchickenthighmeat Theterm“zoneofmaximumicecrystalformation”iswellknownin the field of food freezing and is a temperature zone ranging fromthe freezing point of the food (usually  − 1  ◦ C in the case of fish) to − 5 ◦ C(Yoshikawaandothers1990).However,inthisstudy,weindi-cate the point at which the temperature reached the initial thawing point ( − 5  ◦ C) by using the term “thaw.” Furthermore, in the dura-tion of freezing process, the temperatures were recorded from − 1to − 5 ◦ Cevery10min,andthefreezingpoint(Tk)wasestimatedby the equation Tk  = 0.57 W   − 2.28, where  W   is the weight of chickenmeat dried at 105  ◦ C for 24 h (Goral and Kluza 2002). The thawing  was performed at − 3 and 4  ◦ C, and the thawing curve for chickenthigh meat is shown in Figure 2. As shown in Figure 2, compared tothe conventional thawing method, the chicken thigh meat thawedin an HVEF took less time to reach the initial thaw temperature(from − 16 to − 5  ◦ C). Moreover, when chicken thigh meat reachedthe initial thaw temperature, the thawing time was 40 and 60 minat − 3  ◦ C, respectively. Show the thawing in E-group was more timesaving because the thawing time was 2/3 of a common refrigera-tor (R-group, control). When the thawing temperature was set at4  ◦ C, the thawing time of E-group was 3/4 the time taken for thaw-ing meat using a common refrigerator, and was less than that re-quired for thawing meat at − 3  ◦ C. Moreover, the study by Ohtsuki(1991) who applied this method to electrostatically thawed beef slices indicates that the thawing time for beef slices at  − 3 to 3  ◦ C was only 1/3 to 1/4 the time required for thawing using traditionalmethods, whereas there was no obvious fluid loss after thawing.This was consistent with the results of this study. Microbialqualityofthawedchickenthighmeat Fresh meat is a rich medium for microbial growth, which leadsto spoilage if the meat is not appropriately stored (Nadeem Ahmedand others 2003). Furthermore, the initial microbial content of thefoodaffectsmicrobialgrowthduringstorage.Figure3illustratestheTVC values of frozen chicken thigh meat thawed at  − 3 and 4  ◦ C. Whenthethawingtemperaturewassetat − 3 ◦ C,theinitialTVCval-ues for E- and R-group were 4.3 × 10 3 and 7.0 × 10 3 CFU/g, respec-tively. After 8 d of storage, the TVC values of E- and R-group were2.3 × 10 4 and 2.7 × 10 4 CFU/g, respectively. The reason for the ab-senceofsignificantincreaseintheTVCvaluesandotherpropertiesmight be the ice thawing temperature ( − 3 ◦ C) that was in the rangeofcontrolledfreezing-pointstorage.Controlledfreezing-pointstor-age at nonfreezing temperature zone between the freezing point of  waterandthatofanindividualmaterialcanprolongtheshelflifeof fresh foods and also provide good quality retention. The surround-ing temperature was set between − 0.5 and − 3 ◦ C (Mizuno and oth-ers 1990). When the thawing temperature was set at 4  ◦ C, the initialTVC values for E- and R-group were 3.3 × 10 3 and 4.6 × 10 3 CFU/g,respectively.Onthe6thdofstorageforR-group,theTVCvalueshadalready surpassed the value of 10 7 CFU/g, which was considered astheuppermicrobiologicallimitforgood-qualityfreshpoultrymeatas defined by the Intl. Commission on Microbiological Specifica-tion for Foods (ICMSF 1986). By the 8th d, the microbial counts for Time (minutes) 120100806040200    T  e  m  p  e  r  a   t  u  r  e   (   o    C   ) -18-16-14-12-10-8-6-4-20 Figure 2---Thawing curve of frozen chicken thigh meatthawed at  − 3 and 4  ◦ C.  − 3  ◦ C R-group,  − 3  ◦ C E-group,4  ◦ C R-group, and 4  ◦ C E-group. Error bars represent ±  SD ( n   =  6). Vol. 75, Nr. 4, 2010 —  JOURNAL OF FOOD SCIENCE  M 195  M  :    F     o    o    d    M  i      c   r    o    b    i      o   l      o     g     y    &    S     a   f      e   t       y    Effect of high-voltage electrostatic field on the frozen chicken thigh meat. .. E- and R-group were 4.5 × 10 7 and 4.9 × 10 8 CFU/g, respectively.R-group began to seep mucus and emit foul odors on the 6th dof storage. The spoilage of chicken meat generally occurred whenthe aerobic plate count reached 10 7 to 10 8 CFU/g (Sawaya and oth-ers 1993), which was usually after 4 to 10 d during storage at 4  ◦ C(Jim´enez and others 1997).  Volatilebasicnitrogenmeasurementofthawedchickenthighmeat  Volatile basic nitrogen is an index for evaluating food decay, andit results from degradation of proteins and nonprotein nitrogenouscompounds, mainly as a result of microbial activity (Connell 1975). When the VBN increases rapidly, it implies that the food is be-ginning to decay (Rezaei and others 2007). Figure 4 illustrates the VBN of frozen chicken thigh meat thawed at  − 3 and 4  ◦ C. Whenthe thawing temperature was set at − 3  ◦ C, the VBN content for E-and R-group was 6.55 and 7.40 mg/100 g, respectively. When themeat was stored till the 4th d, the VBN started to increase. By the8th d, the VBN content for the E- and R-group were 19.8 and 19.0mg/100 g, respectively. In both groups, the VBN did not reach the Storage (days) 1086420    l  o  g   C   F   U   /  g 03456789 Figure 3---Changes in total viable counts of frozenchicken thigh meat thawed at  − 3 and 4  ◦ C.  − 3  ◦ C R-group,  − 3  ◦ C E-group, 4  ◦ C R-group, and 4  ◦ C E-group. Error bars represent  ±  SD ( n   =  6). Storage duration (days) 1086420    V   B   N   (  m  g   /   1   0   0  g   ) 051015202530354045 Figure 4---Changes in VBN of frozen chicken thigh meatthawed at  − 3 and 4  ◦ C.  − 3  ◦ C R-group,  − 3  ◦ C E-group,4  ◦ C R-group, and 4  ◦ C E-group. Error bars represent ±  SD ( n   =  6). 20mg/100glimitfordecayanddeterioration(Pearson1968).Whenthe thawing temperature was set at 4  ◦ C, the VBN for E- and R-groupwere10.3and10.5mg/100g,respectively.WhenE-groupwasstored till the 6th d, the VBN content was 14.7 mg/100 g. By the8th d, it was 21.9 mg/100 g. During the time between the 4th and8th d of storage, the VBN of R-group began to show a dramatic in-crease from 16.1 to 39.9 mg/100 g. The meat began emitting odorsand mucus and approached the deterioration level. These resultsindicated that electrostatic storage can inhibit microbial growthand increase storage time. pHvaluechangeinfrozenchickenthighmeatthawedat − 3and4 ◦ C Figure 5 illustrates the pH value of frozen chicken thigh meatthawed at − 3 and 4  ◦ C. When the thawing temperature was set at − 3 ◦ C,thepHvaluesforbothE-andR-grouphadatendencytode-crease at first and then increase subsequently. During the periodof storage between the 2nd and 4th d, the pH value for E-groupfluctuated from 6.78 to 6.56; on the other hand, the pH value forR-group fluctuated from 6.77 to 6.57. During the period of storagebetween the 4th and 8th d, the pH value for E-group fluctuatedfrom 6.56 to 6.77; the pH value for R-group fluctuated from 6.57to 6.62. However, these were considered minor fluctuations. Whenthe thawed temperature was set at 4  ◦ C, the pH values for both E-and R-group were increased with storage time. The pH value forE-group increased from 6.62 to 6.90, whereas that for R-group in-creased from 6.66 to 6.99. Ruiz-Capillas and Moral (2001) and Mas-niyom and others (2002) reported that the increase of pH valuesduring the storage period may be attributed to the production of basic compounds such as ammonia and trimethylamine as well asother biogenic amines. Evaluationofthawingloss,cookingloss,andwaterholdingcapacityinfrozenchickenthighmeatthawedat − 3and4 ◦ C Thesensorialcriteriaarecolor,marbling,anddriploss,bywhichthe consumer is able to judge meat quality (Otto and others 2006).In addition, drip loss and WHC are the most important quality characteristics for the processing industry (Otto and others 2004).The thawing loss, cooking loss, and the WHC of frozen chickenthigh meat thawed at − 3 and 4  ◦ C are shown in Table 1. When the Storage duration (days) 1086420   p   H 6.46.66.87.07.2 Figure 5---Changes in pH of frozen chicken thigh meatthawed at  − 3 and 4  ◦ C.  − 3  ◦ C R-group,  − 3  ◦ C E-group,4  ◦ C R-group, and 4  ◦ C E-group. Error bars represent ±  SD ( n   =  6). M 196  JOURNAL OF FOOD SCIENCE — Vol. 75, Nr. 4, 2010         M      :       F      o      o       d       M       i      c      r      o       b       i      o       l      o      g      y       &       S      a       f      e      t      y Effect of high-voltage electrostatic field on the frozen chicken thigh meat. . . Table 1--- Thawing loss, cooking loss, and WHC of frozenchicken thigh meat thawed at  − 3 and 4  ◦ C. Thawing CookingTreatment loss (%) loss (%) WHC (%) E-Group ( − 3  ◦ C) 0 20.7 ± 0.95 a 89.2 ± 0.56 a R-Group ( − 3  ◦ C) 0 21.5 ± 1.22 a 88.1 ± 1.02 a E-Group (4  ◦ C) 2.9 ± 1.22 a 23.5 ± 2.82 b 86.3 ± 0.75 b R-Group (4  ◦ C) 3.1 ± 0.82 a 26.6 ± 1.11 c 84.7 ± 0.32 c Values represent means ± SD ( n  = 6).Different superscript letters (a, b, c) in the same column indicate significant dif-ference at  P   < 0.05. thawing temperature was set at  − 3  ◦ C, the values of thawing lossfor E- and R-group were almost zero. Moreover, the values of cook-ing loss were 20.7% and 21.5%, whereas the WHC were 89.2% and88.1% for E- and R-group, respectively. The effects of thawing by electrostatic or traditional methods showed no significant differ-ence ( P   >  0.05). Similar results were obtained on microbiologicalquality and quality changes between the 2 treatments. When thethawing temperature was set at 4  ◦ C, the values of thawing loss forE-andR-groupwere2.9%and3.1%,respectively,displayingnosig-nificantdifferences( P  > 0.05).However,asmallbutstatisticallysig-nificant difference ( P  < 0.05) in cooking loss and WHC throughoutthe thawing process was recorded. The values of cooking loss were23.5% and 26.6%, whereas the WHC were 86.3% and 84.7% for E-and R-group, respectively. A similar conclusion was also reportedby Uemura and others (2005) in pork-thawing systems. Conclusions N ovel thawing methods are required for energy conservationand improvement of quality. There are obvious time saving advantages in using HVEF-assisted thawing at ice-cold tempera-ture ( − 3  ◦ C). The time required for thawing to reach the initialthawing point ( − 5  ◦ C) using HVEF treatment was only 2/3 of thatrequired for the R-group. The E-group demonstrated better resultsthan the R-Group ( P  < 0.05) with regard to cooking loss and WHCat 4  ◦ C. Therefore, products thawed using HRS can be expected tohave lower nutritional and physiological losses compared to prod-ucts thawed solely by thermal processing. The HVEF may stimu-latethemotionofwatermoleculesinfood,thuspromotingthawing anddecreasingthethawingrate.Thus,futureinvestigationsforop-timizationofHVEFprocessparameters(electrostaticfieldstrength,thawing temperature) are being planned. Acknowledgment The authors thank the Natl. Science Council of the Republic of China for financially supporting this research under contract nrNSC-95-2313-B-212-0009. References  Anderson BA, Singh RP. 2006. 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