Cumin and Ginger as Antioxidants in Cookies

September 27/Cereal Chemistry -- The effects of cumin and ginger as antioxidants on dough mixing properties and cookie quality were evaluated. Antioxidant activities in cookies were estimated by total phenolic compounds (TPC) contents and free radical scavenging activities. The cookie dough development was evaluated using Mixolab equipment which showed that addition of cumin did not change dough stability and C2, but decreased C3 and C4, while the addition of 5% ginger decreased dough stability (from 8.4 in the control sample to 6.7 min with 5% ginger addition), C2 (from 0.49 in the control sample to 0.31 N-m with 5% ginger addition), C3 and C4.

Cookies formulated with addition of cumin and ginger had increased spread ratios, were softer, and had lower L* and b* values (were darker) than the control. Sensory analysis showed that cookies with cumin and ginger additions had overall acceptability similar to that of the control with a slightly darker appearance, as confirmed by color determination. Using cumin and ginger significantly increased TPC contents from 78.5 in the control to 93.0 and 109.8mg of gallic acid equivalent/100g. respectively.

Similar results were observed in the antioxidant activity measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH), which increased from 41.0% in the control to 51.5% and 64,6%, respectively, for cookies with 5% addition of cumin and ginger.

Recently, antioxidant-supplemented foods have gained a great deal of interest from consumers because of the growing awareness of cancer prevention and the risk of free radicals in the diet. The aim of food producers, food processors and food scientists has been to increase nutritional value of food products supplemented with antioxidants while simultaneously improving formulas for consumer acceptability.

The oxidation of lipids in foodstuffs results in the development of off-flavors, rendering the product unacceptable for human consumption and limiting the shelflife of products. The structure of foods is changed during processing, and as a result, lipids may become more exposed to oxygen. In addition, naturally occurring antioxidant systems are impaired during processing, making processed food more susceptible to oxidation.

Usually, antioxidants such as butylated hydroxytoluene (BHT) and butylaled hydroxyanisole (BHA), both powerful synthetic antioxidants, are used to reduce the rate of oxidation processes. However, these antioxidants suffer from the drawback that they are volatile and readily decompose at high temperatures (Martinez-Tome et al. 2001). Additionally, they are believed to possess carcinogenic activity. These observations have led to a demand for antioxidants derived from naturally occurring sources (Lindberg et al 1995). Many herbs and spices, usually added to season dishes, are an excellent source of phenolic compounds, which have been reported to show good antioxidant activity (Zheng and Wang 2001).

Cumin (Cuminum cyminum (L.), family Apiacea) is one of the most widely used spices in food preparations; it has a warm, bitterish taste and a strong aromatic smell. It is also used in traditional medicine in Arabic countries, India, China, and in Mediterranean countries (Thippeswamy and Naidu 2005). Among the large number of spices used to flavor foods and beverages in India, cumin occupies a place of prominence (Milan et al 2008). It has appreciable amounts of essential amino acids like lysine and threonine (Uma Pradeep et al 1993). The seeds contain a volatile oil (2-5%) that imparts the characteristic aroma to the seeds; the proximate composition of the seeds indicates that they contain oil ([approximate]10%), protein, cellulose, sugar, and other mineral elements (Kim et al 2009). The essential oils found in cumin provided better antioxidative activity when compared to synthetic antioxidants, which could find its application in foods (Allahghadri et al 2010). The seeds are used as an essential ingredient in mixed soups, sausages, pickles, cheese, and meat dishes, and for seasoning breads, cakes and candies (Behera et al 2004).

Ginger (Zingiber officinale (L.) Rose, family Zingiberaceae) has been used as a spice for over 2,000 years (Bartley and Jacobs 2000). Ginger rhizomes and its extracts contain polyphenol compounds (6-gingerol and its derivatives) with a high antioxidant activity (Chen et al 1986). Besides 6-gingerol, ginger contains a homologous series of phenolic ketones expected to have antioxidant effects; such phenolic ketones include 4-, 8-, 10-, and 12-gingerols (He et al 1998).

Several studies on the application of some natural antioxidants to bakery products have been reported. Cookies prepared with rice bran extract (a potential source of antioxidants) had nutritional and health benefits when compared with other commonly used antioxidants. Rice bran made cookies more stable (longer shelflife) that was attributed to its natural antioxidant activity (Bhanger et al 2008). Red palm olein and red palm shortening have been added to cookies with the objective of providing high amounts of natural antioxidant (Al-Saqer et al 2004). Bakery products with the addition of mango dietary fiber showed higher antioxidant capacity associated with extracted polyphenols content (VergaraValencia et al 2007).

In the present study, cumin and ginger powders were used as a source of antioxidants and development of a distinct flavor (increased spiciness) in cookies. The aim of this study was to investigate the properties of cookies prepared with the addition of cumin and ginger. The effects of these spices in the dough properties, antioxidant activity, and consumer acceptability of cookies are reported.

MATERIALS AND METHODS
Low-gluten, soft wheat flour with 1 1.2% protein, cumin and ginger powders, sugar, shortening, salt, and baking soda were obtained from the local markets of Wuxi city, China. All chemicals were of analytical grade. Diphenyl-picrylhydrazyl (DPPH), Folin-Ciocalteu reagent (2N), and gallic acid were purchased, as were acetone, methanol and sodium carbonate.

Moisture and Color
The moisture contents of the flour, flour with additives, and cookies were determined using an air oven according to Approved Method 44- 15 A (AACC International 2010). Flour moisture was determined by the two-stage method while moisture content of cookies was determined using the one-stage method.

Color was determined according to the method of Hyvonen and Slotte (1983) using a chromameter (CR-400, Konica MinoltaSensing, Japan). Color was determined as L*, a*, and b* values, with L* value represents lightness (brightness), a* value represents redness (where +a* is redness while -a* is greenness), and b* value represents yellowness (+b* is yellowness while -b* is blueness). A white-colored calibration plate No. 20533051 (x = 94.0, y = 0.3158, and z = 0.3322) was used to calibrate the machine between every two measurements. Two replicates of every flour sample were analyzed and average value plus or minus standard deviation was reported. Six measurements per cookie sample were measured and the results reported as the average plus or minus the standard deviation of the replicates.

Dough Mixing Property by Mixolab
The Mixolab was used to study the dough mixing properties of control dough (flour alone) and samples with cumin and ginger powder additions. A certain amount of flour with known moisture content was placed into the Mixolab analyzer bowl and mixed to obtain a dough of 75g. The water required for the dough to produce a torque of 1.1 Nm (C1) was added automatically by the Mixolab system. Five parameters (C1, C2, C3, C4, and C5) are obtained from a Mixolab curve (Fig. 1). C1 represents the maximum point of the first mixing stage, while points C2-C5 represent the end points of the corresponding mixing stages. According to manufacturer suggestions, the obtained curve is separated into five stages. In the first stage, dough-mixing characteristics such as stability, elasticity, and water absorption can be determined. During this stage, an increase in the torque is observed until a maximum is reached. Consistency of the dough decreases with mixing, which is an indication of protein weakening (stage 2). As the temperature increases, first a decrease and then an increase in consistency is observed and is attributed to starch gelatinization (stage 3). In stage 4, consistency decreases, attributed to anxiolytic activity. Finally, in stage 5, the decrease in temperature causes an increase in the consistency, attributed to gel formation. This stage is also related to the retrogradation of starch. In this study, we tried to adjust the C1 torque value to 1.1 N-m to establish a comparison between the other Mixolab characteristics. (Roseli et al 2007; Huang et al 2010).

Cookie Preparation and Texture
Cookies were prepared according to Approved Method 10-50D (AACC International 2010) with one modification. In the original method, a dextrose solution is used to aid in developing the brown color. In this study, as cumin and ginger powders have a brown color, the color of cookies did not need dextrose to form a brown color, so water was substituted for the dextrose solution. Cumin and ginger powders were added at 1, 2, 3, 4, and 5g/100g of flour, and the control consisted of flour without any addition. The cookie formulation for control sample and cumin and ginger additions is presented in Table I. Prepared dough was sheeted to 7mm thick and cut using a 54mm diameter circular die. Cookies were baked at 205°C for 11 minutes. Thickness, diameter, and spread ratio were measured 30 minutes after removing the cookies from the oven.

A three-point bend test was made on six cookies using a texture analyzer (TA-TX2, Stable Microsystem, Surrey, England) equipped with a 25kg load cell. The peak breaking force (g) of cookies using the force-in-compression was recorded. Cookie samples were placed on base beams with a distance of 4cm between the two beams. A three-point bending rig was used with an HDP/BS, knife-edge probe. The analyzer was set at a return-to-start cycle, with a pretest speed of 2mm/sec, test speed of 2mm/sec, posttest speed of 10mm/sec, the trigger force was 20g, and distance was 20mm.

Sensory Evaluation Test
In the absence of direct methods to measure taste and aroma, sensory evaluation of cookies provides a practical and rapid test of quality. The results obtained from the sensory-evaluation test enable the evaluation of food and help to judge consumer acceptance without following detailed chemical or microbiological methods. Cookies were presented on white, with disposable plates to 12 panelists using three-code (xxx) labels. The panelists from China, Egypt, Yemen and Pakistan were seven females and five males, aged 22-44 years old with a mean of 33 years, who were asked to rate each sensory attribute. Cookies were evaluated for surface appearance and color, texture, taste, and overall acceptability on a nine-point hedonic scale scoring system: 1, dislike extremely; 5, neither like nor dislike; and 9, like extremely (Hooda and Jood 2005; Zhu et al 2010).

Total Phenolic Compound (TPC) Content
The cookie samples were finely ground by a laboratory blade mill to pass through a 1.0-mm screen. Cookie powder (1g) was extracted using 50mL of methanol at room temperature for 1 hour with continuous swirling using an orbital shaker. Extracts were filtered and stored at -20°C until further analysis for antioxidant determination (Chan et al 2008).

Total phenolic contents of the extracts of flour, flour with cumin and ginger powder addition, and cookie samples were determined using the Folin-Ciocalteu method as described by Emmons et al (1999). Distilled water (4mL) were mixed with 500µL of saturated sodium carbonate, 250µL of sample extract, and 250µL of Folin-Ciocalteu reagent diluted with water (1:1 v/v). The mixture was allowed to stand at room temp for 25 minutes, centrifuged for 10 minutes at 5,000 Ö g at room temp, and absorbance at 725 nm was determined. Results were expressed as mg of gallic acid equivalents (GAE)/100g of sample (mg of GAE/100g).

Antioxidant Activity by Free-Radical Scavenging Activity
DPPH (1,1-diphenyl-2-picrylhydrazyl radical) was used to determine the scavenging activity of the acetone extracts of both flour and cookie samples (control and with cumin powder and ginger powder addition) as described by Tepe et al (2005), with slight modifications. A 2mL aliquot of extract was added to 2mL of DPPH solution (200µM in methanol). The mixture was shaken vigorously and incubated at room temperature in the dark for 30 minutes. The absorbance of the mixture was determined at 517nm using a spectrophotometer. The antioxidant activity was calculated as: where AA is the antioxidant activity, ADPPH is the absorption of the DPPH solution, and A^sub sample^ is the absorption of the extract.

Statistical Analyses
All the experimental data are presented as mean values plus or minus the standard deviation of the mean of the replicates of the individual samples. Data were analyzed for significance using the SPSS 17.0 statistics program, with a one-way analysis of variance (ANOVA) performed at the significance level of 0.05%. Duncan's multiple range test was used to differentiate between mean values, and standard deviation was calculated using the same software.

RESULTS AND DISCUSSION
Characteristics of Flour with Addition of Cumin and Ginger
The moisture content of control flour and flour with cumin and ginger additions were presented in Table II. Moisture content was 13.6, 13.4, and 13.2% for flour, cumin, and ginger powders, respectively.

The control flour, cumin and ginger powders, and flours with different addition levels were analyzed for color. For cumin powder, the respective values were 55.5, 4.28, and 18.68 for L*, a*, and b*; and for ginger powder the respective values were 72.09, 5.59, and 24.14 for L*, a*, and b* values (Table II). Adding cumin and ginger powder to the flour gave a darker, higher a* and b* values, which was the reason for removing dextrose solution from the cookie formula. A dextrose solution was added to the original formula to aid browning on the surface of cookies. The modification of the cookie formula was made by substituting water for the dextrose solution.

The lightness values of the flour samples gradually decreased with increasing addition levels of cumin and ginger powders (P = 0.05) (Table II). Lightness decreased by 2.5 and 3.2% with 5 and 10% addition levels, respectively, compared to the control. The lightness of cumin additions was significantly higher than the ginger additions (P = 0.05). The lowest value was obtained by 10% addition, with a lightness value of 93.1. Addition of cumin powder did not affect the redness values significantly, while ginger powder addition to the flour, increased a* values significantly. The highest value was achieved by the maximum addition level of ginger powder in the 10% addition, with an increase of 133.3% compared to the control. The b* values of the flours showed the same trend of a* values. The b* values were not affected significantly by the addition of cumin powder, while with addition of ginger powder, the b* values increased significantly. Maximum b* value was obtained by the maximum addition of ginger powder at 10%, with a difference of 32.7% compared to the control.

Mixolab Parameters
Adding cumin and ginger powders did not significantly affect water absorption, dough development time, and C5 (P = 0.05), while the stability, C2, C3, and C4 values were significantly different (P = 0.05) (Table III). Addition of cumin powder (1-5%) did not affect the dough stability of tested flour samples significantly compared to the control sample, while the addition of ginger powder (1-5%) decreased the dough stability significantly compared to the control sample. It was interesting to see the different effects between cumin and ginger additions and between different levels. That may be because different levels of components such as fiber or phenolic compounds are in different additions. The exact reason was unclear and justifies further research.

C2 is attributed to a weakening of protein, with lower C2 values corresponding to lower protein quality at increased temperature (Anonymous 2005). Cumin additions did not change C2 compared to the control sample, while ginger additions showed lower C2 values than the control. The maximum change was observed with the maximum addition of ginger (5%), which decreased C2 from 0.49 to 0.31 N-m. The differences between C1 and C2 (C1 - C2) were related to the diameter of cookies, which is an indicator for protein quality. The greater the difference between C1 and C2 values, the lower the protein quality (Ozturk et al 2008). C1 - C2 was greater in ginger-added flours. The diameter of ginger-added cookies was larger than the control and larger than cumin- added cookies. Cumin additions had greater C1 - C2 than the control sample and the diameter of cookies with cumin additions were larger than the diameter of the control sample. C3 decreased gradually from 2.05 N-m in the control to 1.87 and 1.97 N-m in 5% additions of cumin and ginger, respectively. C4 was also decreased from 1.99 Nm in the control to 1.81 and 1.9 Nm in 5% additions of cumin and ginger, respectively. The lowest values for C3 and C4 were observed in cumin additions at 5%.

Cookies Characteristics
The moisture contents of the cookies are presented in Table IV. No significant differences were observed with additions of cumin and ginger powders, except for cumin additions at 4 and 5%, which had higher moisture contents compared to the control, and with cumin addition at 5% having the maximum value of 8.19%. The high moisture content could be due to the high content of total dietary fiber in cumin powder, enabled to hold more water than the low-gluten flour. Cumin has a total dietary fiber content (TDF) of 59.0%, insoluble dietary fiber (IDF) content of 48.5%, and soluble dietary fiber (SDF) content of 10.5%, which all act to increase the water-holding capacity (Sowbhagya et al 2007).

It is well-known that the color of cookies is one of the most important quality attributes that influence acceptability for consumers. The surface color of prepared cookies from all additions was analyzed using the same method as for determining the color of the flours (Minolta Hunter Lab color space). The values obtained are presented in Table II. Cumin and ginger powder additions caused a gradual decrease in L* values, resulting in darker cookies compared to the control. Differences between cumin and ginger powder additions were not highly significant (P = 0.05). Our findings were similar to those reported by Ajila et al (2008), who found that increasing the mango-peel powder level decreased the L* value.

Adding cumin decreased a* values significantly (P = 0.05). In ginger powder additions, the a* value of the cookie surfaces was higher than the control. The highest value was obtained in 4% ginger addition, which was 18.5% higher than control. Both cumin and ginger powders addition decreased the b* values significantly (P = 0.05). Percentage decreases were 6.4 and 5.1% for 5% addition of cumin and ginger, respectively, compared to the control. No significant differences between cumin additions and ginger additions were seen. Cookie diameter, thickness, and spread ratio were measured 30 min after removing cookies from the oven, according to Approved Method 10-50D (AACC International 2010). Physical parameters are shown in Table IV. Cumin and ginger powders addition affected diameter, thickness, and spread ratio significantly (P 0.05). The cookie diameter increased significantly (P = 0.05) from 40.3 cm for the control to 42.5 and 42.4 cm for 5% of cumin and ginger additions, respectively, while the thickness decreased from 7.43 cm for the control to 6.5 and 6.58 cm for 5% of cumin and ginger additions, respectively. As a result of the changes in diameter and thickness, the spread ratios increased significantly (P = 0.05) with the addition to the cookie formula of both cumin and ginger powders at different levels. The values increased from 5.43 for the control to 6.54 and 6.44 for 5% of cumin and ginger additions, respectively.

The firmness of the cookies was evaluated by the breaking force tests and are presented in Table IV. The data show that cookie samples were significantly affected by the use of cumin and ginger powders in the formulation. The firmness (breaking force) decreased proportionally with the increase in the percentages of both cumin and ginger powders, but ginger affected the breaking force more than cumin. The maximum percentage reduction in force was obtained by the maximum addition level in ginger addition at 5%, with a decrease of 28.5% compared to the control sample; 5% addition of cumin decreased the breaking force by 21.2% compared to the control, which was 4.7kg. Cookie samples prepared with the addition of cumin and ginger were softer than the control (lower breaking force) but with significantly differences between them.

Sensory Evaluation
Cookies produced with cumin or ginger powder had good overall acceptability scores without significant differences from the control. This suggests that the products would have acceptability by the consumers. Table V shows the sensory evaluation scores given by the 12 panelists. Cumin additions scored lower for appearance by the panelists, which was in correlation with the color determination in Table II. In cumin additions, the L*, a*, and b* values were lower compared to the other additions and the control sample. This was confirmed by the results obtained from the panelists. However, the highest level of cumin and ginger additions (5%) had scores of 6.8 and 7.5. not significantly different from the control sample with an appearance score of 8.

The texture and flavor of cookies with cumin and ginger additions were scored as acceptable, with no significant differences (P = 0.05) from the control sample. Texture scores showed a trend to lower values than the control, confirming what was found in the firmness test (Table IV), which showed that both cumin and ginger additions resulted in cookies with lower breaking force compared to the control. However, the panelist scores were not statistically significant. This indicates that cookies with cumin and ginger powders additions had similar sensory attributes (texture, flavor, and overall acceptability) compared to the control sample. Therefore, prepared cookies, even with cumin and ginger additions at the highest level of 5% are expected to have good acceptability.

Total Phenolic Compounds (TPC) Contents in Cookies
Folin-Ciocalteu is not considered to be an authentic antioxidant test, but it provides an indication of the total phenolic compounds of the extracts and this is related to antioxidant activity because most of the plant-derived antioxidants consist of polyphenols (Lloycd et al 2000). Total phenolic compounds (TPC) of the methanolic extracts from the cookie samples are summarized in Table VI. The TPC contents of cumin and ginger powders were 566 and 1,132mg gallic acid equivalent (GAE)/ 100 g of dry sample, respectively. The additions of cumin and ginger powders influenced the TPC contents of the cookies significantly (P = 0.05). The control sample had the lowest content with a TPC level of 78.49 mg GAE/100 g of dry sample. Ginger additions resulted in higher TPC than cumin additions, with 5% ginger addition yielding an increase of 39.9% TPC compared to the control. In comparison, with 5% cumin addition the TPC level was 18.5% higher than the control.

Determined values of TPC of cookie samples were less than the calculated values. That might be due to the heat instability of TPC during baking process. The results were in line with findings from previous researches (Lindberg et al 1995; Mansour and Khalil 2000; Cheng et al 2006: Li et al 2007).

Free Radical Scavenging Activities of Cookies
DPPH (1,1-diphenyl-2-picrylhydrazyl) is a stable radical of organic nitrogen, characterized by a deep purple color and a maximum absorbance in the range of 515-520 nm (Locatelli et al 2009). DPPH is one of a few stable and commercially available radicals that can be used to determine antioxidant activity. Antioxidant compounds react with DPPH (1,1-diphenyl-2-picrylhydrazyl). converting it into 2,2-diphenyl-1-picrylhydrazyl. The extent of the decrease in absorbance at 517 nm indicates the scavenging potential of the antioxidant extract, which is due to the hydrogendonation ability of the antioxidant compound (von Gadow et al 1997). The scavenging of the stable DPPH radical has been used widely to evaluate the antioxidant activity of phenolic-compound extracts of cereal grains (Lin et al 2009). fruits (de Oliveira et al 2009). and herbs (Wojdylo et al 2007).

DPPH scavenging activity data are presented in Table VI. The ginger additions showed higher DPPH scavenging than cumin additions, and both had a higher scavenging capacity compared to the control sample, which scavenged 40.6% of the DPPH radical. The addition of cumin and ginger powders at 5% to the cookie formula increased scavenging of the DPPH radical to 51.5 and 64.6%, respectively. Increasing the levels of both cumin and ginger powders gradually increased the ability of the methanolic extracts to scavenge the DPPH radical. Thus, the addition of cumin and ginger powders to cookies yielded products with a significant source of antioxidant capacity. These results comfirm that such a relatively small percentage of cumin and ginger can improve the perceived value of a very well-accepted bakery product with enhanced nutraceutical properties. There are a number of spices with known bioactive compounds that have to be studied to probe the residual benefit after processing has been applied and the final product analyzed.

CONCLUSIONS
Adding cumin and ginger powders to flour at levels of 1-5g/ 100g of flour improved antioxidant activity as measured by both Folin-Ciocalteu and DPPH assays. TPC was 1 8 and 40% higher than the control when 5% of cumin and ginger, respectively, were added. DPPH scavenging activity was increased from 22.45% in the control to 51.48 and 64.57%, respectively, in cookies with added cumin and ginger at the 5% addition levels. Ginger additions showed higher TPC and DPPH-scavenging activity than control and cumin additions. Limited changes in dough development properties were observed with the addition of cumin and ginger and the effects did not significantly affect the end product, as shown by the sensory evaluation. The color of cumin cookies was darker than the control, while ginger cookies have similar colors to that of the control. All cookies had acceptable scores and were similar to the control. These products can be commercially produced, thus offering options of convenience and "better for you" bakery products for present day consumers.

ACKNOWLEDGMENTS
We are grateful for the financial support of research from the National Natural Science Foundation of China (20576046 and 307705) from the China Food Industry Group, Nanjing, China, and from Fortune Bakery Co., Ltd, Zhangjiagang, China, and by the earmarked funding for the Modern Agro-industry Technology Research System (No. nycytx-14) from the Ministry of Agriculture of China.

Mohamed Abdel-Shafi Abdel-Samie1 Jingjing Wan, 1  Weining Huang, 1,2 Okkyung Kim Chung, 1  and Baocai Xu3
1  Graduate researcher, professor, and visiting professor, respectively. The State Key Laboratory of Food Science and Technology, International Exchange and Cooperation Program. School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214036, China.
2  Corresponding author. Phone: +86-510-8591 9139. Fax: +86-510-8591 9139. E-mail: wnhuang@jiangnan.edu.cn
3  Research scientist. China Food Industry Group. Nanjing. Jiangsu 210041, China.

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From the October 4, 2010, Prepared Foods E-dition