Pandesal) Using Hydrocolloids and Emulsifiers

Philippine Journal of Science 145 (1): 25-35, March 2016 ISSN 0031 - 7683 Date Received: ?? Feb 20??

Staling Control in Philippine Yeast Bread (Pandesal) Using Hydrocolloids and Emulsifiers

Maria Patricia V. Azanza1,2*, Emil Emmanuel C. Estilo1, and Florenda S. Gabriel3

1Department of Food Science and Nutrition, College of Home Economics, University of the Philippines Diliman, Quezon City, Philippines 1101 2Industrial Technology Development Institute, Department of Science and Technology, Bicutan, Taguig City, Philippines 1631 3Department of Home Economics Education, College of Home Economics, University of the Philippines Diliman, Quezon City, Philippines 1101

The short 3-day shelf-life of Philippine yeast bread (Pandesal) was extended by controlling staling and mold growth with antimicrobials, hydrocolloids, and emulsifiers, singly or in combination. Addition of combined antimicrobials 0.30% (flour basis, fb) calcium propionate and 0.10% (fb) potassium sorbate in a reference basal Pandesal recipe controlled mold growth up to 5 d, but did not delay earlier onset of staling (4 d). Reformulations of the basal recipe with combined antimicrobials using the hydrocolloids pectin and xanthan gum (0.25% and 0.50% fb levels each) were able to control bread firming up to 5-6 d in addition to mold growth control. Incorporation of hydrocolloids produced denser breads marked by increased weight, specific volume, and moisture content. Treatment of 0.50% (fb) pectin of bread formulation with antimicrobials yielded the best results in terms of overall acceptability and longest shelf-life, and was used in the subsequent reformulation with emulsifiers. Addition of monoacylglycerol (MAG) and sodium stearoyl lactylate (SSL) (0.25% and 0.50% fb levels each) further delayed firming up to 7 d with mold growth generally limiting the shelf-life of Pandesal. Incorporation of emulsifiers also improved bread volume and produced softer crumbs with 0.25% MAG yielding the best results. The compounded additives of 0.30% (fb) calcium propionate, 0.10% (fb) potassium sorbate, 0.50% (fb) pectin, and 0.25% (fb) MAG were found best to extend Pandesal use-by date to a total of 7 d.

Key words: bread, pandesal, pectin, staling, xanthan gum

INTRODUCTION Philippine food culture was considered as the result of the introductions of wheat flour as an ingredient and baking Pandesal or Philippine salt bread, basically composed as a mode of cooking by Spanish and other foreign settlers of wheat flour, sugar, salt, shortening, and yeast, is in the country (Fernandez & Best 2000). The defining considered as the traditional breakfast bread staple in characteristic of Pandesal from other local breads is the the country (Guzman et al. 1986; Dagoon 2005; Albala salt added to the dough as well as the use of breadcrumbs 2011). Breads that are leavened with carbon dioxide gas after molding and panning. The breadcrumbs are ultimately produced by yeast are also known as yeast breads (Luna responsible for the rough surface texture of the Pandesal 2005; Brown 2010). Indigenization of bread in the local crust after baking. A 100 g edible portion of Pandesal has *Corresponding author: [email protected] an estimated energy value of 330 kcal and a proximate

25 Philippine Journal of Science Azanza et al.: Staling Control in Pandesal Vol. 145 No. 1, March 2016 composition of 21.6 g moisture, 10.1 g crude protein, 4.2 quality (Rosell et al. 2001, Sharadanant & Khan 2003, g crude fat, 62.9 g available carbohydrates, 6.0 g dietary Guarda et al. 2004). Pectin has been reported to affect fiber, and 1.2 g ash (FNRI 1997). Unfortunately, the local water mobility (Sun-Waterhouse et al. 2011) and influence micro- to small-scale bakeries were reported to usually the development of the gluten network (Baik & Chinachoti have only 2-3 d ambient shelf-life for this traditional 2000). Xanthan gum has been reported to improve dough staple (Mayo 2000, Tumimbang 2008). Shelf-spoilage of handling characteristics (Collar et al. 1999), which could Pandesal could be addressed at two levels of deterioration: confer a greater stability of the gluten-starch network microbial and staling. Pandesal, like any bread stored at in the composite dough during baking (Shittu et al. ambient conditions, is very prone to mold spoilage. Mold 2009). This study aimed to address the short shelf-life spoilage of breads is mainly attributed to contamination of Pandesal by controlling mold growth through the use from spores in the environment surrounding the breads of antimicrobials and textural deterioration through the during cooling, slicing, packaging, and storage (Pateras application of both hydrocolloids and emulsifiers. 2007). Considering the commercial viability of Pandesal in the Philippines, there is still paucity in information related to the control of its deterioration as a result of mold growth and staling. MATERIALS AND METHODS Staling is a textural deterioration phenomenon that also limits shelf-life of bread generally manifested by an Bread ingredients and additives increase in crumb firmness followed by flavor and aroma Commercial bread flour, salt, sugar, shortening, instant deterioration (León et al. 1997; Palacios et al. 2004). Crust dry yeast, and eggs were acquired from the local staling is generally caused by moisture transfer from the retail market. The food grade antimicrobials calcium crumb to the crust (Lin & Lineback 1990), resulting in a propionate and potassium sorbate were procured from soft, leathery texture, and is generally less objectionable RTC Laboratory Services and Supply House, Manila, than crumb staling (Pateras 2007). Crumb staling is more Philippines. The hydrocolloids used in the study were complex, more important, and less understood (Short pectin (Pectin from RTC Laboratory Services and & Roberts 1971). León et al. (1997) have described the Supply House, Manila, Philippines) and xanthan gum changes that undergo in the starch during bread baking (Ziboxan F200 from FUDynamics International Inc., and cooling. These included the gelatinization process Manila, Philippines). Local distributors provided the that involves increase of granule volume due to starch emulsifiers used in the study: sodium stearoyl lactylate hydration, disruption of granule structure, heat absorption, (SSL) (Lecinta Plus from Bakels Philippines, Manila, and loss of granule crystallinity. Upon slow cooling of Philippines) and monoacylglycerol (MAG) (Ovalett from starch paste, retrogradation process occurs as described by Bakels Philippines). gel formation resulting to an increased crystalline order of starch molecules (Ward et al. 1994, DeMan 1999). Several Preparation of Pandesal samples authors have associated the staling phenomenon with A straight dough process was used for preparing a the retrogradation process, particularly of amylopectin reference basal commercial Pandesal formula based molecules of the starch present during baking and storage on 3000 g flour weight from the procedure used in (Palacios et al. 2004, Zhou et al. 2008). the University Food Service Bakeshop, University of Kohajdová et al. (2009) have studied the effect of different the Philippines, Diliman, Quezon City. The following emulsifiers to slow down starch retrogradation. Generally, reference formulation was used on a flour basis (fb): emulsifiers such as diacetyl tartaric acid esters of mono- 25% water, 1% instant dry yeast, 20% sugar, 1% salt, and diglycerides of fatty acids (DATA ESTERS, DATEM), 10% shortening, 5.4% eggs. Dough was optimally mixed, sodium stearoyl-2-lactylate (SSL) and monoacylglycerols fermented for 15 min and divided into approximately 60-g (MAG) can delay retrogradation by either its interaction rolls before being panned and proofed for 45 min at 30°C. with the amylose and amylopectin or binding with water, Baking was carried out for 15 min at 150°C prior to 30-min making it unavailable to participate in gel formation cooling at ambient temperature. Breads were packaged in (Cauvain & Young 2003; Kohajdová et al. 2009). HDPE bags (Calypso, 0.030 mm total thickness, Calypso Plastic Center Co., Binondo, Manila, Philippines) and Aside from emulsifiers, hydrocolloids are also considered stored at ambient conditions (29.9±0.7°C, 70.9±6.6% RH) as one of the additives for delaying starch retrogradation. monitored with a digital meter (Traceable® Humidity/ Schiraldi et al. (1996) and Davidou et al. (1996) Thermometer/Clock Monitor, Control Company, Texas, investigated the use of hydrocolloids as anti-staling agents USA). and demonstrated their softening ability. Among those, pectin and xanthan gum have been used to improve bread

26 Philippine Journal of Science Azanza et al.: Staling Control in Pandesal Vol. 145 No. 1, March 2016

Additive incorporation of Fiszman et al. (2005) was created through panel Additive incorporations were done in accordance to the consensus. Presentation and familiarization of panelists provisions and limits dictated by the Codex Alimentarius with reference standards and textural attributes were (FAO and WHO 2011) and the Philippine Food and carried out prior to sensory analysis (Table 1). Crust Drug Administration (FDA Philippines 2006). Test quality was based on graininess while crumb quality was antimicrobials, hydrocolloids, and emulsifiers were based on soft central area, hardness, oral cohesiveness, sequentially selected and cumulatively incorporated. and mechanical cohesiveness. The reference basal formula was incorporated with the following antimicrobials: 0.30% (fb) calcium propionate Panelists were given Pandesal samples coded with three- and 0.10% (fb) potassium sorbate. For hydrocolloid digit random numbers and were tasked to evaluate by addition, pectin and xanthan gum were independently comparing with established reference standards. Spider incorporated at levels of 0.25% (fb) and 0.50% (fb) in plots were created in order to describe the intensity profiles breads with antimicrobials. For emulsifier addition, of the textural attributes. In the acceptability tests, a SSL and MAG were similarly incorporated at levels 7-point bipolar Hedonic scale (ranging from dislike very of 0.25% (fb) and 0.50% (fb) each to the basal recipe much to like very much) was used to determine the degree with antimicrobials along with the best kind and level of of acceptability for each textural attribute evaluated and hydrocolloid. Sequential selection of the type and level the overall acceptability. Scores for the acceptability tests of additive was based on its capacity to produce breads were considered as one of the parameters in determining that remained acceptable for the longest possible period the end of shelf-life of Pandesal. while being free from mold growth. Sequential recipe reformulations were carried out by subtracting the weight Yeast and mold count of the additives from the flour weight while keeping other The yeast and mold count (YMC) of Pandesal was ingredients constant. determined according to procedures described in the Bacteriological Analytical Manual (Tournas et al. -5 Technological and physicochemical evaluation 2001). Serially diluted samples of up to 10 were pour- Procedures used for the technological evaluation of plated using acidified Potato Dextrose Agar (HiMedia bread described by Guarda et al. (2004) included Laboratories Pvt. Ltd., Mumbai, India) in duplicates. determinations for: weight, volume, specific volume, Counts were reported as log colony forming units (cfu)/g and width/height ratio of the central dorsoventral lateral sample. slice (1 cm thick from the center of the bread). Moisture content determination was performed using a moisture Shelf-life evaluation analyzer (MAC 50/NH, Radwag, Radom, Poland). A water End of shelf-life was based on staling [Es] and/or mold activity meter (ms1 Set aw, Novasina, Switzerland) and growth [Em]. Sensory acceptability and intensity scores a pH meter (pH 700 Bench Meter, Eutech Instruments, based on textural parameters were used as indices of Singapore) were used to measure respective water activity staling. The date when the overall acceptability rating falls

(aw) and pH values of bread samples. Bread firmness below 4 (less than neither like nor dislike) was determined was evaluated by determining the compression rates as the [Es] date, while the date for observed mold growth of Pandesal samples using a penetrometer (H-1200, was determined as the [Em] date. The day before the

Humboldt, Humboldt Mfg. Co., Chicago, USA) modified established [Es] and/or [Em] was identified as the use-by to simulate a compressibility meter. With a compression date of the control and additive-modified Pandesal. time of 15 s, trials were done on the left edge, center, and right edge surfaces of the bread and measurements Statistical analysis were reported as compression rates expressed in mm/sec. Data obtained from all independently replicated (n=3) Results were reported as mean value ± standard deviation experiments were subjected to single-factor Analysis based on three trials. of variance (ANOVA) using IBM SPSS Statistics 21 software (IBM Corp., 2012, New York, USA). The results Sensory evaluation of analyses were presented as mean ± standard deviation. Descriptive analysis and acceptability tests were Duncan’s Multiple Range Test (DMRT) was used as post- simultaneously used to characterize the staling process hoc analysis when a significant difference existed among in Pandesal. A panel of 5 (female, age group of 22-28 means (at 5% level of significance). years old) were trained for determining intensity ratings of the textural attributes measuring the degree of staling in Pandesal based on Descriptive Analysis (Meilgaard et al. 1999). A modified lexicon derived from the study

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RESULTS AND DISCUSSION End of shelf-life of Pandesal made from the reference commercial basal recipe was attributed to both molds and staling. This coincides with the claims of local Panaderos Acceptability Ratings (bakers) from micro- to small-scale bakeries that their Antimicrobial Effects. Table 2 shows the mean sensory Pandesal usually deteriorates 2 d after baking at ambient ratings for the overall acceptability and crust and crumb storage. With the addition of calcium propionate at 0.30% acceptability ratings of breads prepared from the reference (fb) and potassium sorbate at 0.10% (fb) in Pandesal, basal commercial Pandesal recipe and antimicrobial- microbiological shelf-life was extended by 2 d as mold modified formulation. The antimicrobial combination of spoilage was controlled. Extension of the mold-free shelf- additives used was 0.30% (fb) calcium propionate and life of bread by the addition of calcium propionate has 0.10% (fb) potassium sorbate. Entries highlighted in gray also been reported by Williams and Pullen (2007). Despite (Table 2) showed the identified use-by date preceding the the extended microbiological shelf-life, the use-by date end of shelf-life based on staling [E ] and/or observed s was however observed earlier at 4 d as Pandesal became visible mold growth [E ]. m unacceptable due to staling. The control Pandesal was observed to have a use-by date Weak acid preservatives such as calcium propionate and of 3 d, while Pandesal with antimicrobial treatment had potassium sorbate, in their undissociated state, act on a use-by date extended up to 4 d with staling preceding microbial cells by penetrating through the membrane. mold spoilage. No adverse effects in fresh bread sensory Substrates and enzymes are subsequently inactivated and characteristics after incorporation of antimicrobials were eliminated by lowering the intracellular pH through the observed. Both freshly-baked control and antimicrobial- ionization of acid molecules and the release of hydrogen treated Pandesal exhibited the same overall acceptability ions (Jay 1992; Lallemand 1996; Theron & Lues 2011). score of near like moderately. Moreover, no significant Although breads fresh out of the oven were reported to be difference existed (p=0.256) between the acceptability free of molds or mold spores due to thermal inactivation scores of the freshly-baked samples in both treatments during baking (Ponte Jr. & Tsen 1987), post-processing for all other textural parameters of the crumb and crust. contamination is usually the most common source of mold Staling due to crumb firming was mostly observed in spoilage (OTA 1979, Pateras 2007). the fresh Pandesal of both the control and antimicrobial- Hydrocolloid Effects. The addition of hydrocolloids treated breads after a day in storage as most changes in was able to generally improve the shelf-life and control crumb textural parameters were already significantly staling as use-by date was extended by at most 2 d (Table different (p<0.001, data not presented). The staling 3). Incorporation of hydrocolloids at 0.25% (fb) was process is known to result in extensive textural and sensory able to extend use-by date up to 5 d while incorporation changes such as crumb hardening and crust softening at 0.50% (fb) was able to further extend use-by date (Piazza & Masi 1995). Rapid loss of quality due to staling up to 6 d. Pectin incorporation generally increased the begins just when breads are taken out from the oven overall acceptability (like moderately) of freshly baked (Zeleznak & Hoseney 1986). These changes may involve breads while xanthan gum produced breads with slightly several physical and chemical phenomena including the lower acceptability scores (near like moderately) when recrystallization of the amylose and amylopectin starch compared to the antimicrobial-treated bread (Table 2). components (Krog et al. 1989, Zobel & Kulp 1996), both No significant difference (p=0.172) was also observed the loss and redistribution of water (Zeleznak & Hoseney among the acceptability scores of freshly-baked breads 1986, Biliaderis 1992) and the protein-starch interactions in all hydrocolloid treatments for all crumb and crust (Martin et al. 1991). Increase in crumb firmness has textural parameters. Similarly, Lazaridou et al. (2007) been the textural attribute used to the largest extent by also reported positive acceptability scores (between like investigators following bread staling (D'Appolonia & slightly and like moderately) of bread supplemented with Morad 1981; Gray & Bemiller 2003). hydrocolloids, including pectin and xanthan gum at test In other staling studies, bread crust has also been observed levels of 1.0 and 2.0% (fb). It was observed, however, to stale and develop a leathery texture as water migrates that several Pandesal units with 0.50% (fb) xanthan gum from the crumb to the crust during storage (Goesaert et al. exhibited fractures in the bread crust after baking (data 2009; Delcour & Hoseney 2010). However, results from not shown). Guarda et al. (2004) reported that breads this study have shown that crust did not seem to be affected that were made with a basic straight dough recipe using by staling as sensory scores for the graininess of the crust commercial wheat flour gave the lowest acceptability were still acceptable even at the end of shelf-life. Perhaps, scores in appearance after being supplemented with 0.10% breadcrumbs may have imparted a masking effect in the (fb) xanthan gum, with the values being even worse at sensory perception of crust staling of Pandesal. high concentrations (0.50% fb).

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During storage, crumb hardening was observed as interface between gluten and starch, thus improving gas acceptability scores for all crumb parameters in all retention (Krog 1981, Kokelaar et al. 1995). It has also hydrocolloid-treated breads decreased significantly been reported that SSL interaction with gluten proteins (p<0.001) throughout storage. Delay in crumb firming during mixing caused gluten aggregation and increased was best observed in bread with 0.50% (fb) pectin dough strength (Gómez et al. 2004). followed by 0.50% (fb) xanthan gum, as all textural parameters were still acceptable up to 6 d. Staling The emulsifier MAG, known to be a crumb softener, was manifested earlier in breads with 0.25% (fb) levels of reported to retard the firming process based on its ability to pectin and xanthan gum incorporation as some textural form complexes with amylose (Stampfli & Nersten 1995). parameters were only acceptable up to 5 d. Nonetheless, Specifically, the part of the amylose which is known to hydrocolloid incorporation was able to delay crumb complex with MAG has been reported to not participate firming as acceptability scores decreased at a lower rate in the gel formation which normally occurs with the starch compared to the antimicrobial-treated breads (Table 2). in unmodified dough during baking, making it unable to Increasing hydrocolloid incorporation was also observed recrystallize and contribute to staling of the bread crumb to further extend shelf-life of Pandesal, with 0.50% (fb) upon cooling (Stampfli & Nersten 1995). pectin incorporation producing the highest acceptability The modified Pandesal with antimicrobials and 0.50% ratings. It has been proposed that the beneficial effects of (fb) pectin incorporated with the lesser concentration of hydrocolloids in delaying staling result from a combined MAG (0.25% fb) was selected to establish the complete opposite phenomenon (Biliaderis et al. 1997). One of combination of additives as it had produced the highest the observed effects of hydrocolloid incorporation in overall acceptability scores. Again, it was observed that bread was reported to be an increase in structural rigidity graininess did not deteriorate immediately relative to the attributed to the decreased swelling of the starch granules crumb parameters. and reduced leaching of amylose. The other effect was the weakening of the composite starch bread network that could also be observed due to the inhibition of Intensity Ratings interaction among swollen granules. It was also noted in Figure 1 presents the intensity ratings for the crust and this study that aside from controlling staling, the addition crumb parameters of freshly-baked breads and those of xanthan gum and pectin further delayed mold growth at their identified use-by dates. It can be observed that with increasing concentration of hydrocolloid. sequential and cumulative incorporation of additives extended the use-by date to a total of 7 d from the 3 d Emulsifier Effects. The addition of emulsifiers in of the reference basal formula. Intensity profiles of the Pandesal modified with antimicrobials and 0.50% (fb) spider plots of the different freshly-baked breads tend to pectin generally had further increased the use-by date by exhibit the same characteristics: grainy crust, very chewy another 1 d (Table 4). Moreover, all treatments were able and cohesive crumb with a very soft central area. With to improve fresh bread characteristics as all parameters addition of antimicrobials, the use-by date was extended obtained scores higher than those just containing from 3 d to 4 d, as cohesiveness (both oral and mechanical) antimicrobials and 0.50% (fb) pectin (Table 3). Staling still and soft central area continued to decrease, while hardness proceeded at storage although at an apparent slower phase. increased even without visible mold growth up to 4 d Emulsifiers are believed to control staling because of the of storage. However, the decrease in these fresh bread following phenomena: these become trapped by the gluten textural parameters were less pronounced with the addition phase during dough mixing of breads, then released toward of pectin as breads at the use-by date were relatively more the starch gel during baking, after which these would cohesive and had a softer central area. Pectin (0.50% fb) remain primarily in the intergranular regions where they and MAG (0.25% fb) were able to decrease the crumb can form complexes with leached amylose or amylopectin textural deterioration of Pandesal with antimicrobial (Chinachoti & Vodovotz 2001). Emulsifiers are known control up to 7 d. to improve the final quality characteristics of bread as these have been reported to improve gas retention and Technological Evaluation dough handling by increasing dough strength and stability Antimicrobial Effects. The results of the evaluation of the (Gray & Bemiller 2003; Gómez et al. 2004) and increase technological parameters of the control, antimicrobial- dough mixing tolerance (Azizi & Rao 2004). For SSL, treated Pandesal, and formulations with the selected increasing its concentration generally improved crumb hydrocolloid and/or emulsifier (Table 5) showed that textural scores, which was similarly reported by Alasino et with incorporation of antimicrobials, reduction in specific al. (2011) in wheat breads. The effect of SSL was reported volume can be observed in the breads. However, as to be attributed to its action as a dough strengthener; it is indicated by the values of the width/height ratio, bread capable of forming liquid films of lamellar structure at the

29 Philippine Journal of Science Azanza et al.: Staling Control in Pandesal Vol. 145 No. 1, March 2016 configuration generally did not change significantly rates as an indicator of bread firming could be attributed in (p=0.117). Likewise, no significant difference was also part to the crystallization of amylopectin due to moisture observed between the means of specific volume (p=0.057) movement within the crumb (Goesaert et al. 2009). Piazza and moisture content (p=0.086) for freshly-baked control & Masi (1995) stated that staling brings about both crust and the antimicrobial-modified Pandesal. softening and crumb hardening. According to Hasan & Abdolgader (2012), calcium Hydrocolloid Effects. Significant changes (p<0.001) propionate is considered an effective inhibitor for rope were observed on technological bread parameters bacteria and molds with little or no effect on yeast such as specific volume, moisture, compression rate, population. At normal use rates (0.20% fb), calcium aw and pH upon addition of 0.50% (fb) pectin to the propionate has been reported to have minimal or no antimicrobial-treated Pandesal. The increase in moisture effect on flavor as well as the leavening activity of yeast content of freshly-baked Pandesal along with a decrease

(Beuchat & Golden 1989; Lallemand 1996; Pateras in aw observed in the study could be attributed to the 2007). However, Legan (1993) reported a 3-5% volume reduced amount of free water due to the hydrophilic reduction in laboratory-scale and 5-10% in commercial- nature of hydrocolloids (Nussinovitch 1997). Moreover, scale baking of standard British white loaves after using Trombetta et al. (2005) reported that the nature of a 0.20% (fb) calcium propionate. Sorbates were also hydrocolloid or protein polymer network and their reported to have even greater adverse effects attributed crosslinking mechanisms, which is likewise applicable to the decreased leavening activity of yeasts and the in the crumb structure of breads, has been reported to dough becoming sticky and difficult to process (Legan reduce water activity. Hydrocolloids improve water 1993). Most preservatives that inhibit bacteria are also retention in breads when used in small quantities (<1% capable in inhibiting yeast. Accepted theories on their fb in flour) (Kohajdová et al. 2009) and are also able to action suggest inhibition through internal pH depression modify gluten and starch properties that could delay the by directly inhibiting glycolysis enzymes (Guynot et al. retrogradation process (Collar et al. 1999; Bárcenas et 2004). Some practices to alleviate this effect include using al. 2009; Kohajdová et al. 2009). Moisture content of relatively acid-resistant yeast, or increasing the amount bread plays an important role in crumb firming. Rogers of yeast in the formulation, or by applying sorbates only et al. (1988); Xu et al. (1992) and Hug-Iten et al. (2003) topically after baking (Lallemand 1996). found that the moisture content was inversely proportional to the rate of firming. He & Hoseney (1990) similarly Freshly-baked control Pandesal significantly (p<0.001) reported that the higher the moisture content, the slower had lower pH values relative to additive-modified breads. the firming rate and the lower the final firmness of bread. Dissociation of salts due to hydrolysis in aqueous solutions Schiraldi & Fessas (2001) proposed that, since water acts will produce their respective acids and bases (Whitten as a plasticizer in the bread, greater hardness is yielded et al. 2014). In this study, the dissociation of potassium when the decrease in the moisture content favors the sorbate and calcium propionate into weak acids and formation of hydrogen bonds among the starch polymers strong bases might account for the slight increase in pH or between the starch and the proteins. Armero & Collar observed in the antimicrobial-modified breads. Softer (1997) proposed that the weakening effect on the starch crumb of the control was established and had a relatively structure due to hydrocolloid addition promotes better higher compression rate compared to the antimicrobial- water distribution and retention and a decrease in the treated samples. As staling progressed, crumb firming was crumb resistance. Pectin has also been reported to affect observed through changes in the technological parameters gluten hydration as it is able to induce a decrease in the of both treatments that were clearly manifested by the swelling of gluten and increase its water-binding capacity significantly decreasing (p<0.001) compression rates. (Gray & Bemiller 2003; Bárcenas et al. 2009). According to Goesaert et al. (2009), moisture moves from the crumb to crust during storage, and seemingly, Specific volume and width/height ratio of freshly-baked within the crumb from gluten to the slowly crystallizing Pandesal were observed to decrease, which was similarly amylopectin. This moisture loss by the crumb to the crust observed by Sun-Waterhouse et al. (2011) after treating is said to be responsible for the increase in crumb firmness. wheat breads with pectin at varying levels. It has been reported that the addition of pectin has been known to For the antimicrobial-treated samples, specific volume improve dough strength due to its strong water-binding was observed to decrease along with storage period. No capacity (Correa et al. 2012), which could negatively significant difference was observed among the means of other affect dough expansion during proofing. Throughout technological parameters (p=0.117 for width/height ratio, storage, the most obvious trend with hydrocolloid addition p=0.086 for moisture content, p=0.119 for compression rate, could be seen in the significantly decreasing (p<0.001) p=0.100 for water activity, p=0.082 for pH) within treatments values for compressibility. Although there was an initial as storage period increased. The decreasing compression

30 Philippine Journal of Science Azanza et al.: Staling Control in Pandesal Vol. 145 No. 1, March 2016 increase in the compression rates, subsequent firming was (0.30% fb calcium propionate, 0.10% fb potassium sorbate), still observed. Nonetheless, the effect of hydrocolloid hydrocolloid (0.50% fb pectin), and emulsifier (0.25% fb addition on decreasing the aw could possibly help explain MAG), the initial microbial load was decreased by about 2 the added effect in the extension of the microbiological log cycles. Control and additive-modified Pandesal tend to shelf-life of Pandesal by 1 d. have YMC at about 4.5-5.0 log cfu/sample at their use-by dates. With or without additives, it was noted that the onset Emulsifier Effects. Addition of MAG at 0.25% (fb) in of mold growth in Pandesal was observed when the YMC Pandesal with 0.50% (fb) pectin and antimicrobials yielded approaches values of about 5 log cfu/g bread. significant increases (p<0.001) in the moisture content, compression rate and aw of freshly-baked breads along Breads in general were reported to typically have a with delayed staling (Table 5). Incorporation of 0.25% (fb) shelf-life of 2 d under ambient conditions by the Office MAG was able to improve bread specific volume, although of Technology Assessment of the United States (OTA the changes were not significant (p=0.162). Emulsifiers 1979). Results of this study confirmed the reports of are generally known to improve gas cell retention through Panaderos from local micro- to small-scale bakeries strengthening the gluten network (Carr et al. 1992; Gan et that their Pandesal usually deteriorates 2 d after baking al. 1995). The emulsifier MAG has been found to improve at ambient storage based on staling and mold growth. bread crumb structure and volume through the stabilization The combined use of antimicrobials calcium propionate of the liquid lamellae which separate gas cells in the dough (0.30% fb) and potassium sorbate (0.10% fb) have (Carr et al. 1992; Gómez et al. 2004). Junge et al. (1981) shown to effectively delay mold spoilage of the reference stated however that some emulsifiers such as MAG do Pandesal. Moreover, this study also showed that the not affect the amount of air occluded during mixing, but addition of hydrocolloids and emulsifiers did not only rather result to a higher number of smaller cells that may retard staling but microbiological growth as well. be formed and stabilized. This resulting fine crumb grain can be attributed to increased air incorporation during dough mixing, to smaller cells formed during mixing, or to a combination of both (Pareyt et al. 2011). CONCLUSION Incorporation of 0.25% (fb) MAG also significantly Shelf-life deterioration of the Philippine yeast bread improved (p<0.001) water retention and crumb softness Pandesal can be observed on 2 aspects: microbiological than the reference Pandesal with 0.50% (fb) pectin and and staling. The study was able to verify the claim of local antimicrobials (Table 5). The crumb softening effect of Panaderos regarding the use-by date of Pandesal under MAG has also been previously documented (Stampfli & ambient Philippine storage conditions of 2-3 d without the Nersten 1995; Gómez et al. 2004; Sawa et al. 2009), which amendment of additives. The combined use of calcium is attributed to their interaction with starch during baking. propionate (0.30% fb) and potassium sorbate (0.10% fb) This interaction with starch granules decreases their ability was able to extend the microbiological shelf-life by an to absorb water and swell, creating a softer crumb structure additional 2 d but staling still manifested through textural (Krog 1981; Collar et al. 1999). Several bread staling crumb firming. The use of breadcrumbs was observed to studies have reported an inverse relationship between the possibly mask the perception of crust staling in stored rate of firming and moisture content (Rogers et al. 1988; Pandesal. Intensity profiles from descriptive analysis He & Hoseney 1990; Xu et al. 1992; Hug-Iten et al. 2003). showed that freshly-baked Pandesal has a grainy crust Breads with 0.25% (fb) MAG exhibited moisture content and a very chewy and cohesive crumb with a very soft that was significantly higher (p<0.001) among other central area. As staling progressed, sensory profile of treatments which could account for their longer shelf- Pandesal at the use-by date indicated breads that still life. The adsorption of emulsifiers onto starch granule have a grainy crust but with a relatively firmer and less surfaces as well as complex formation was believed to chewy crumb. The hydrocolloid pectin at 0.50% (fb) and prevent starch from taking up water released from gluten emulsifier MAG at 0.25% (fb) were found to effectively during bread aging (Krog 1981; Rao et al. 1992). Perhaps delay the staling process in Pandesal. Mold control was this moisture barrier mechanism of emulsifiers was also also observed as pectin and MAG incorporation was responsible for the observed extension of microbiological able to further extend microbiological shelf-life each by shelf-life of Pandesal by 1 d. 1 d. Overall, the established complete combination of additives extended the use-by date of Pandesal from 3 d to a total of 7 d. Microbiological Analysis Table 6 shows that freshly-baked control Pandesal Sensory evaluation methods, based on acceptability already have YMCs of about 4.0 log cfu/g sample. With tests and descriptive analysis, were shown to be a very the sequential and cumulative addition of antimicrobials important tool in this study in determining the shelf-life

31 Philippine Journal of Science Azanza et al.: Staling Control in Pandesal Vol. 145 No. 1, March 2016 of Pandesal. The appreciation of sensory evaluation as a BILIADERIS CG. 1992. Structures and phase transitions means to evaluate the shelf-life of breads should still be of starch in food. Journal of Food Technology verified in conjunction with more objective technological 46(6):98-109. methods that include state-of-the-art equipment and BILIADERIS CG, ARVANITOYANNIS I, techniques in the evaluation of bread. Likewise, given IZYDORCZYK MS, PROKOPOWICH DJ. 1997. simple and appropriate terminologies associated with Effect of hydrocolloids on gelatinization and structure sensory evaluation, bakery personnel and bread consumers formation in concentrated waxy maize and wheat starch would have a better understanding of data generated from gels. Starch 49(7-8):278-283. shelf-life studies. BROWN AC. 2010. Understanding Food: Principles and Preparation. California, USA: Wadsworth, Cengage Learning. 704p ACKNOWLEDGEMENTS CARR NO, DANIELS NW, FRAZIER PJ. 1992. Lipid The authors would like to express their gratitude to interactions in breadmaking. Critical Reviews in Food the Department of Science and Technology – National Science and Nutrition 31(3):237-258. Capital Region (DOST-NCR) for funding this project. CAUVAIN SP, YOUNG LS. 2003. Water control in The authors would likewise acknowledge the University baking. In: Bread Making: Improving Quality, Cauvin Food Service, UP Diliman, Quezon City for the assistance SP, Editor. Florida, USA: Woodhead Publishing in the preparation of Pandesal. Due credit is also given to Limited. p.447-466 Ms. Alpha Grace Legaspi for extending their help in the conduct of the study. CHINACHOTI P, VODOVOTZ Y. 2001. Bread Staling. New York, USA: CRC Press LLC. 192p COLLAR C, ANDREU P, MARTÍNEZ JC, ARMERO E. 1999. Optimization of hydrocolloid addition to REFERENCES improve wheat bread dough functionality: a response ALASINO MC, OSELLA CA, DE LA TORRE MA, surface methodology study. Food Hydrocolloid, SANCHEZ HD. 2011. Use of sodium stearoyl lactylate 13(6):467-475. and azodicarbonamide in wheat flour breads with CORREA MJ, PÉREZ GT, FERRERO C. 2012. Pectins added pea flour. International Journal of Food Sciences as breadmaking additives: Effect on dough rheology and Nutrition 62(4):385-391. and bread quality. Food and Bioprocess Technology ALBALA K. 2011. Food Cultures of the World 5(7):2889-2898. Encyclopedia. California, USA: ABC-CLIo. 1400p DAGOON J. 2005. Home Economics Technology IV ARMERO E, COLLAR C. 1997. Texture properties (First ed.). Manila, Philippines: Rex Printing Company, of formulated wheat doughs - Relationships with Inc. 214p dough and bread technological quality. Zeitschrift für D'APPOLONIA BL, MORAD MM. 1981. Bread staling. Lebensmittel-Untersuchung und -Forschung 204(2): Cereal Chemistry 58(3):186-190. 136-145. DAVIDOU S, LE MESTE M, DEBEVER E, BEKAERT AZIZI MH, RAO GV. 2004. Effect of surfactant gels on D. 1996. A contribution to the study of staling of dough rheological characteristics and quality of bread. white bread: effect of water and hydrocolloid. Food Critical Reviews in Food Science and Nutrition 44(7- Hydrocolloids 10(4):375-383. 8):545-552. DELCOUR JA, HOSENEY RC. 2010. Principles of BAIK M, CHINACHOTI P. 2000. Moisture redistribution Cereal Science and Technology (3rd ed.). Minnesota, and phase transitions during bread staling. Cereal USA: AACC International, Inc. 270p Chemistry 77(4):484-488. DEMAN JM. 1999. Principles of Food Chemistry (3rd ed). BÁRCENAS ME, DE LA O-KELLER J, ROSELL CM. Maryland, USA: Aspen Publishers, Inc. 520p 2009. Influence of different hydrocolloids on major wheat dough components (gluten and starch). Journal FERNANDEZ DG, BEST J. 2000. Palayok: Philippine of Food Engineering 94(3-4):241-247. Food Through Time, On Site, In The Pot. Manila, Philippines: The Bookmark, Inc. 123p BEUCHAT L, GOLDEN D. 1989. Antimicrobials occurring naturally in foods. Food Technology FISZMAN SM, SALVADOR A, VARELA P. 2005. 43(1):134-142. Methodological developments in bread staling

32 Philippine Journal of Science Azanza et al.: Staling Control in Pandesal Vol. 145 No. 1, March 2016

assessment: application to enzyme-supplemented HUG-ITEN S, ESCHER F, CONDE-PETIT B. 2003. brown pan bread. European Food Research & Staling of bread: Role of amylose and amylopectin Technology 221(5):616-623. and influence of starch-degrading enzymes. Cereal Chemistry 80(6):654-661. [FAO] Food and Agriculture Organization & [WHO] World Health Organization. 2011. Codex General JAY JM. 1992. Modern Food Microbiology, 4th ed. New Standard for Food Additives (GSFA) Online Database. York: Van Nostrand Reinhold. 701p Retrieved from http://www.codexalimentarius.net/ JUNGE RC, HOSENEY RC, VARRIANO-MARSTON gsfaonline/index.html;jsessionid=DB7C59469C7009 E. 1981. Effect of surfactants on air incorporation in E12EFFF835FF04B47C on 25 February 2013. dough and the crumb grain of bread. Cereal Chemistry [FDA] Food and Drug Administration Philippines. 2006. 58(4):338-342. Updated List of Food Additives. Manila: Department KOHAJDOVÁZ, KAROVIČOVÁ J, SCHMIDT Š. 2009. of Health. 231p Significance of emulsifiers and hydrocolloids in bakery [FNRI] Food and Nutrition Research Institute. 1997. industry. Acta Chimica Slovaca 2(1):46-61. Food Composition Tables. Manila: Food and Nutrition KOKELAAR JJ, GARRITSEN JA, PRINS A. 1995. Research Institute. 163p Surface rheological properties of sodium stearoyl-2- GAN Z, ELLIS PR, SCHOFIELD JD. 1995. Gas cell lactylate (SSL) and diacetyl tartaric esters of mono (and stabilisation and gas retention in wheat bread dough. di) glyceride (DATEM) surfactants after a mechanical Journal of Cereal Science 21(3):215-230. surface treatment in relation to their bread improving abilities. Colloids and Surfaces A: Physicochemical GOESAERT H, SLADE L, LEVINE H, DELCOUR JA. and Engineering Aspects 95(1):69-77. 2009. Amylases and bread firming – an integrated view. Journal of Cereal Science 50(3):345-352. KROG N. 1981. Theoretical aspects of surfactants in relation to their use in breadmaking. Cereal Chemistry, GÓMEZ M, DEL REAL S, ROSELL CM, RONDA F, 58(3):158-164. BLANCO CA, CABALLERO PA. 2004. Functionality of different emulsifiers on the performance of KROG N, OLESEN SK, TOERNAES H, JOENSSON T. breadmaking and wheat bread quality. European Food 1989. Retrogradation of the starch fraction in wheat Research & Technology 219(2):145-150. bread. Cereal Foods World 34(3):281-285. GRAY JA, BEMILLER JN. 2003. Bread staling: LALLEMAND. 1996. A Guide to Baking Preservatives. Molecular basis and control. Comprehensive Reviews Lallemand Baking Update 1(5). Retrieved from in Food Science and Food Safety 2(1):1-21. http://www.lallemand.com/BakerYeastNA/eng/PDFs/ LBU%20PDF%20FILES/1_5 on 4 June 2014. GUARDA A, ROSELL CM, BENEDITO C, GALOTTO MJ. 2004. Different hydrocolloids as bread improvers LAZARIDOU A, DUTA D, PAPAGEORGIOU M, and antistaling agents. Food Hydrocolloids 18(2):241- BELCN, BILIADERIS CG. 2007. Effects of 247. hydrocolloids on dough rheology and bread quality parameters in gluten-free formulations. Journal of Food GUYNOT M, RAMOS A, SANCHIS V, MARIN S. 2004. Engineering 79(3):1033-1047. Study of benzoate, propionate, and sorbate salts as mould spoilage inhibitors on intermediate moisture LEGAN JD. 1993. Mould spoilage of bread: The problem bakery products of low pH (4.5–5.5). International and some solutions. International Biodeterioration and Journal of Food Microbiology 101(2):161-168. Biodegradation 32:33-53. GUZMAN MP, CLAUDIO VS, DE LEON SY. 1986. LEÓN A, DURÁN E, BENEDITO DE BARBER C. 1997. Basic Foods for Filipinos. Manila: Merriam and A new approach to study starch changes occurring in Webster, Inc. 455p the dough-baking process and during bread storage. Zeitschrift für Lebensmittel-Untersuchung und HASAN SM, ABDOLGADER RA. 2012. Study of weak -Forschung 204(4):316-320. acid perservatives and modified atmosphere packaging (MAP) on mold growth in modal agar system. Food LIN W, LINEBACK OR. 1990. Changes in carbohydrate and Nutrition Sciences 3(6):802-809. fractions in enzyme-supplemented bread and the potential relationship to staling. Starch 42(10):385-394. HE H, HOSENEY RC. 1990. Changes in bread firmness and moisture during long-term storage. Cereal LUNA MV. 2005. Guzman’s Introduction to Food Chemistry 67(6):603-605. Preparation: a manual of Laboratory Procedures and Principles in Food Preparation (6th ed.). Manila,

33 Philippine Journal of Science Azanza et al.: Staling Control in Pandesal Vol. 145 No. 1, March 2016

Philippines: Merriam and Webster. 314p NM, PRESTON KR. 2009. Effects of purified monoglycerides on Canadian short process and MARTIN ML, ZELEZNAK KJ, HOSENEY RC. 1991. A sponge and dough mixing properties, bread quality mechanism of bread firming I: Role of starch swelling. and crumb firmness during storage. Food Chemistry Cereal Chemistry 68(1):498-503. 115(3):884-890. MAYO DR. 2000. Storage Study on Pan De Sal Made SCHIRALDI A, FESSAS D. 2001. Mechanism of From Wheat-Coconut Flour Composite. Manila: Staling: An Overview. In: Bread Staling. Chinachoti College of Home Economics. 224p P, Vodovotz Y, Editors. New York, USA: CRC Press MEILGAARD M., CIVILLE GV, CARR BT. 1999. LLC. p.1-17. Sensory Evaluation Techniques, 3rd ed. New York, SCHIRALDI A, PIAZZA L, BRENNA O, VITTADINI USA: CRC Press. 387p E. 1996. Structure and properties of bread dough and NUSSINOVITCH A. 1997. Hydrocolloid Applications: crumb. Journal of Thermal Analysis 47(5):1339-1360. Gum Technology in the Food and Other Industries. SHARADANANT R, KHAN K. 2003. Effect of London: Blackie Academic & Professional. 354p hydrophilic gums on the quality of frozen dough II: [OTA] Office of Technology Assessment. 1979. Bread Characteristics. Cereal Chemistry 80(6):773- Application of Open Dating to Specific Foods. 780. Retrieved from http://www.princeton.edu/~ota/ SHITTU TA, AMINU RA, ABULUDE EO. 2009. disk3/1979/7911/791112.PDF on 13 September 2013. Functional effects of xanthan gum on composite PALACIOS HR, SCHWARZ PB, D'APPOLONIA B. L. cassava-wheat dough and bread. Food Hydrocolloids 2004. Effects of alpha-amylases from different sources 23(8):2254-2260. on the firming of concentrated wheat starch gels: SHORT AL, ROBERTS EA. 1971. Pattern of firmness Relationship to Bread Staling. Journal of Agricultural within a bread loaf. Journal of the Science of Food and Food Chemistry 52(19):5987-5994. Agriculture 22(9):470-472. PAREYT B, FINNIE SM, PUTSEYS JA, DELCOUR JA. STAMPFLI L, NERSTEN B. 1995. Emulsifiers in bread 2011. Lipids in bread making: Sources, interactions, making. Food Chemistry 52(4):353-360. and impact on bread quality. Journal of Cereal Science 54(3):266-279. SUN-WATERHOUSE D, SIVAM AS, COONEY J, ZHOU J, PERERA CO, WATERHOUSE GI. 2011. PATERAS IM. 2007. Bread Spoilage and Staling. In: Effects of added fruit polyphenols and pectin on the Technology of Breadmaking (2nd ed.) Cauvain SP & properties of finished breads revealed by HPLC/ Young LS, Editors. New York, USA: Springer Science LC-MS and Size-Exclusion HPLC. Food Research + Business Media, LLC p.275-298. International 44(9):3047-3056. PIAZZA L, MASI P. 1995. Moisture redistribution THERON MM, LUES JF. 2011. Organic Acids and Food throughout the bread loaf during staling and its effect Preservation. Florida: CRC Press. 340p on mechanical properties. Cereal Chemistry 72:320- 325. TOURNAS V, STACK ME, MISLIVEC PB, KOCH HA, BANDLER R. 2001. Bacteriological Analytical PONTE JR JG, TSEN CC. 1987. Bakery Products. In: Manuel: Yeasts, Molds and Mycotoxins. Retrieved Food and Beverage Mycology, 2nd ed. Beuchat LR. from http://www.fda.gov/Food/FoodScienceResearch/ New York, USA: Van Nostrand Reinhold p.191-223. LaboratoryMethods/ucm071435.htm on 25 June 2013. RAO PA, NUSSINOVITCH A, CHINACHOTI P. TROMBETTA G, DI BONA C, GRAZI E. 2005. The 1992. Effects of selected surfactants on amylopectin transition of polymers into a network of polymers. recrystallization and on recoverability of bread crumb International Journal of Biological Macromolecules, during storage. Cereal Chemistry 69(6):613-618. 35(1-2):15-18. ROGERS D, ZELEZNAK K, LAI C, HOSENEY R. 1988. TUMIMBANG M. 2008. Pandesal, Cookies and Extruded Effect of native lipids, shortening, and bread moisture Snack Curls with Coconut Flour. Retrieved from http:// on bread frming. Cereal Chemistry, 65(5):398-401. www.fnri.dost.gov.ph/index.php?option=content&task ROSELL CM, ROJAS JA, BENEDITO DE BARBER C. =view&id=1432 on 25 June 2012. 2001. Influence of hydrocolloids on dough rheology WARD KE, HOSENEY RC, SEIB PA. 1994. and bread quality. Food Hydrocolloids 15(1):75-81. Retrogradation of amylopectin from maize and wheat SAWA K, INOUE B, LYSENKO E, EDWARDS starches. Cereal Chemistry 71(2):150-155.

34 Philippine Journal of Science Azanza et al.: Staling Control in Pandesal Vol. 145 No. 1, March 2016

WHITTEN KW, DAVIS RE, PECK ML, STANLEY GG. 2014. Chemistry (10th ed.). California: Brooks/Cole Cengage Learning. 1184p WILLIAMS T, PULLEN G. 2007. Functional Ingredients. In: Technology of Breadmaking, 2nd ed. Cauvain SP & Young LS, Editors. New York, USA: Springer Science+Business Media, LLC. p.45-80. XU A, CHUNG OK, PONTE JR JG. 1992. Bread crumb amylograph studies I: Effects of storage time, shortening, flour lipids, and surfactants. Cereal Chemistry 69(5):495-501. ZELEZNAK KJ, HOSENEY RC. 1986. The role of water in the retrogradation of wheat starch gels and bread crumb. Cereal Chemistry 63:407-411. ZHOU Y, WANG D, ZHANG L, DU X, DU X. 2008. Effect of polysaccharides on gelatinization and retrogradation of wheat starch. Food Hydrocolloids 22(4):505-512. ZOBEL HF, KULP K. 1996. The staling mechanism. In: Baked Goods Freshness, Hebeda RE, Zobel HF, Editors. New York, USA: Marcel Dekker, Inc. p.1-64.