Wheat flour extraction rate of 72% was obtained from the South Cairo Mill Company, Al Giza, Egypt. Yellow corn flour 97 % was obtained from Egyptian Company for Maize Products, 10th Ramadan City, Egypt. Carob pods and baking ingredients: dry yeast, cane sugar, corn oil, baking powder, and salt were obtained from local market, Al Giza, Egypt. All chemicals were used in the experiment were obtained from Egyptian Scientific Company, El-Dokki, Al Giza, Egypt.
Methods
Preparation of roasted carob bean powder from carob pulp
Carob bean pods were manually cleaned, freed of impurities, and washed with running water. The seeds were separated and kept away for the extraction of the gum. The obtained pulp was roasted in a convection oven Memmert, Cambridge, UK) at 130° C for 30 minutes according to Boublenza et al. [2], then ground to obtain fine powder passed through a 125 µm sieve. Finally, the obtained powder was kept in polyethylene pages at -18° C for further use.
Extraction of carob bean gum from carob bean seeds
Carob bean gum was extracted from carob bean seeds using a hot water method as described by El Batal, & Hasib [15] with few modifications. Carob bean seeds (100 g) were dehusked after soaking in 800 ml boiling water for 2 h to facilitate the mechanical discarding of the husks and germs. The obtained endosperms were dried in a convection oven Memmert, Cambridge, UK) at 40° C for 18 hrs. The dried endosperms were added to distilled water in a ratio of 1: 197and the pH was adjusted to 8 with NaOH (1M). Then the suspension was submitted to a water bath at 97º C, under shaking (450 rpm), for 36 min. Solid to liquid separation was done by centrifugation at 21875 rpm/4° C for 1h. The residue was discarded, and an equal volume of isopropanol was added to the supernatant to precipitate the carob bean gum. The solution was kept for 18 h at 4 ºC, and carob bean gum was skimmed off and rinsed with isopropanol. After drying under vacuum at 30°C for 18 h, the precipitate was ground to a fine powder and kept in polyethylene pages at -18° C for further use.
Preparation of cupcakes
Formulations presented in Table 1 were used to produce cupcakes according to the method described by Doweidar [16]with some modifications, where cane sugar was replaced by roasted carob powder at 0, 25, 50, and 75 %. Briefly, egg and vanilla were whipped, then corn oil was added to the egg-vanilla mixture gradually and well beaten at low speed for 5 min. After that, cane sugar / roasted carob powder/ cacao powder was added, well beaten at low speed for 5 min. Finally, the other dry ingredients were added to the mixture and blended with a hand mixer for 2 min at high speed. Cupcake batter was poured into greased mini cupcake pans and baked at 180°C for 25min. The cupcakes were allowed to cool, packed in polypropylene bags, and stored for further analyses.
Preparation of gluten-free bread
Formulations presented in Table 2 were used to produce gluten-free bread according to the method described by Zahran [17] with some modifications. Corn flour was mixed with dry yeast, corn oil, and salt. Carob bean gum was dispersed in half of the water to be used in dough formulation by using a high-speed homogenizer. Then the dispersion was heated at 80° C for 15 min within continuous mixing and added to the corn flour mixture. The rest of the water was added and the dough was rested at 30° C/ 15 min/ 85% relative humidity, then divided into portions each of 125 g. The divided portions were flattened to 3 mm thickness and fermented at 30° C / 25 min/ 85% relative humidity. Finally, the fermented dough is baked at 350-400° C for 1-1.25 min. gluten-free bread was allowed to cool before sensory scoring.
Sensory evaluations
Sensory evaluation was carried out on cupcakes according to Larmond [18]. Members from Food Technology Research Institute, Agricultural Research Centre scored cupcakes for taste, odor, appearance, crust color, crumb color, crumb texture, and overall acceptability (OAA).on a 9 hedonic scale from one (dislike extremely) to nine (like extremely). Sensory evaluation was carried out on gluten-free bread according to Johnson & Harrisn[19]. Members from Food Technology Research Institute, Agricultural Research Centre scored the bread for general appearance, crust color, texture, taste, odor, roundness, layer separation, and overall acceptability (OAA). The scoring was done on a 9 hedonic scale from one (dislike extremely) to nine (like extremely).
Chemical analysis for raw materials, cupcakes, and gluten-free bread
Chemical composition was performed on raw materials, cupcakes, and gluten-free bread. The contents of moisture, crude protein, crude fat, ash, fiber, and acid value were determined according to the AACC International [20] Approved Methods 44- 16, 46-30, 30-10,08-01,02-31.01, respectively. Crude protein content was calculated as N × 5.7, and Total carbohydrates was calculated by difference.
The energy(Kcal)was calculated according to equation 1:
Determination of total phenolic content (TPC)and antioxidant activity (AOA)
The TPC content of carob powder before and after roasting was determined according to Singleton,& Rossi [21] using a Folin-Ciocalteu reagent. Samples were measured at 765 nm, and the concentration was expressed as milligrams of Gallic acid equivalents (GAE)/100 g. The AOA of carob powder before and after roasting was evaluated by measuring free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH˙) scavenging capacity according to Blois [22]. The absorbance was measured against pure methanol at 515 nm, and the AOA was calculated as percent discoloration from the following equation:
where A0 is the absorbance of the pure methanol control at the beginning of the reaction (t = 0) and A1 is the absorbance of sample extract at the end of the reaction(t = 30 min). All chemical tests were averaged from three replicates.
Measurement of the Viscosity (η) of the carob bean gum
The dispersion of 1% carob bean gum in distilled water under vigorous stirring for 1 h, at room temperature, followed by heating the dispersion at 85°C in a water bath for 30 min according to Garcia‐Ochoa, & Casas [23]. The apparent viscosity (η, cP) of carob bean gum dispersion was measured using Brookfield Engineering labs, DV-III Ultra Rheometer, and SC4-21 spindle. The shear rate was varying from 9.30 - 55.80/s, and the viscometer was operated between 10 and 60 RPM.
Physical characteristics of the cupcake and gluten-free bread
Water activity(aw)
The aw of the cupcakes and gluten-free bread was measured using Rotronic Hygrolab 3 CH-8303, Switzerland as mentioned by Cadden [24].
Color measurements
The color of the cupcakes and the crust of the gluten-free bread were measured in triplicate using a colorimeter (CR-400, Konica Minolta Sensing Inc., Japan). The color values were recorded as L* =lightness (0 = black, 100 = white), a* (-a* = greenness, +a* = redness) and b* (-b* = blueness, +b* = yellowness).
Textural Profile analysis (TPA)
The TPA of cupcakes and gluten-free bread were measured using Brookfield Engineering Lab. Inc., Middleboro, MA. 02346-1031. USA based on AACC International [20]. method 74-09. From the force-time curves, Firmness/ Hardness (N), Adhesiveness (N.s-1), and Resilience were calculated.
Statistical analysis
The obtained results for chemical composition and sensory evaluation were statistically analyzed by the least significant difference value (LSD) at P ≤ 0.05levels probability by Snedecor & Cochran [25].