Determination of different spice powder levels for soft cheese based on milk

This study aims to evaluate and determine the most optimal level of addition spice powder to soft cheese based on milk. The spice powders used are cinnamon (Cinnamomum burmannii), lemongrass (Cymbopogon citratus), and turmeric (Curcuma longa L.) with the following treatment; T0: soft cheese without added spices powder; T1: soft cheese + 1% cinnamon powder; T2: soft cheese + 2% cinnamon powder; T3: soft cheese + 3% cinnamon powder; T4: soft cheese + 1% lemongrass powder; T5: soft cheese + 2% lemongrass powder; T6: soft cheese + 3% lemongrass powder; T7: soft cheese + 1% turmeric powder; T8: soft cheese + 2% turmeric powder; T9: soft cheese + 3% turmeric powder. The study design used a completely randomized design with three replications. Variables measured including pH, color (L*, a*, b*), total solids, and antioxidant activities using/via DPPH scavenging activity (2,2-diphenyl-picrylhydrazyl). The soft cheese pH ranged from 5.60–6.26, with T0 was the lowest and T3 had the highest value. Total solids ranged from 38.87–66.77%, with T1 was the lowest and T3 had the highest value. Antioxidant activities ranged from 37.96–72.43%, with T0 was the lowest and T9 had the highest value. Soft cheese showed L* value 66.58–80.31, with T9 was the lowest and T0 had the highest value; a* value 3.47–12.44, with T6 was the lowest and T9 had the highest value; b* value 12.35–25.95, with T0 was the lowest and T9 had the highest value. Results were analyzed using analysis of variance and Duncan’s Multiple Range Test. Based on the results, it can be concluded that the level addition of 3% spice powder was the most optimal level for soft cheese, with the highest pH, total solids, antioxidant activity, redness, and yellowness.


Introduction
Consumers want delicious, nutritious, and healthful food products, known as functional foods.Demand for functional foods is increasing worldwide.Cheese is a fermented milk-based food product that is produced in various flavors and forms worldwide [1].Cheese contains substances that have particular biological activities and can be used as functional foods.[2].Functional food is a part of the human diet and has been demonstrated to provide health benefits and decrease the risk of chronic diseases beyond those provided by adequate nutrition [3].The fortification of dairy foods with spices could provide functional dairy products with medicinal and nutritional value [4].The development of cheese as a functional food product includes the use of spices rich in antioxidants, such as cinnamon (Cinnamomum burmannii), lemongrass (Cymbopogon citratus), and turmeric (Curcuma longa L.).These species have the advantage of being a source of antioxidants, including tannins, flavonoids, and steroids.Cinnamon is known to possess antioxidant, antibacterial, and other therapeutic properties [5].Leaves of lemongrass plants are known to have various biological properties, including antioxidant properties [6].Curcumin is the main chemical compound in turmeric and has been shown to have antioxidant activities [7].Spices added to soft cheese may alter the quality of the product, including pH, total solids, color, and antioxidant activity.Spices are added to cheese making to give it a variety of qualities, including color.Cinnamon, lemongrass, and turmeric have different colors.The cinnamon is brown, lemongrass is green, and turmeric is yellow.These three spices are inexpensive, plentiful, and are used as medicines or food in traditional society.
Cheese, kefir, and yoghurt, can be enhanced by using spices.Throughout history, spices have been used as preservatives, flavors, and therapeutic agents to fortify foods.Enhancing cheese aroma, color, flavor, increasing the shelf life of cheese are some of the objectives of using cinnamon, lemongrass, and turmeric powder [8].Spices contain natural antioxidants, that can reduce oxidative cell damage [9].
The spices used in this study were powdered, which possibly affect the color and acidity of the cheese.These qualities were affected by the amount of spice powder used.This study aims to determine and evaluate the most optimal level of addition spice powder to soft cheese based on milk and its effect on the pH, total solids, color, and antioxidant activity.

Spice powder manufacture
The spice powder was manufactured using cinnamon, lemongrass, and turmeric, cut into 2 cm pieces, and dried using a Filter Pro Dehydrator for 12 hours 40 o C.After drying, the spices were ground using a herb grinder (model IC-06B).Finally, the spices were sieved thrice with size of sieve 20 cm and stored in jars containing silica gel.The process of spice powder manufacture can be observed in Figure 1.

Cheese production
The process of cheese production starts with heating 85 o C whole cow's milk for 15 seconds, then allowed cool down to 40 o C in batches, with each batch containing 1 liter of milk.The subsequent step was adding CaCl2 0.2 gram and starter mesophilic 0.01 gram and incubated for 45 minutes 37 o C in an incubator.The function of CaCl2 is to shorten coagulation time and increase curd strength.Pure calf rennet 0.3 mL was added, and kept at 37 o C in incubator for 1 hour (coagulation process).The curd was cut into 2×2 cm.The next step was to separate the whey and curd, and the curd that was formed was added to the spice powder according to the treatment, then hung overnight using a cheese cloth to separate the whey.The separated curd and whey were then pressed using a cheese mold for 1 hour and the curd position was reversed every 30 minutes.Finally, the soft cheese was covered with 2% table salt and stored in a refrigerator for 24 hours before measurement.The process of spices cheese production can be observed in Figure 2.

Determination of antioxidant activity
Soft cheese was prepared for spectrophotometric analysis of the antioxidant activity of DPPH (2,2diphenyl-picrylhydrazyl) [11]. 1 gram of cheese was dissolved methanol 5 mL.After dissolution, the cheese was centrifuged for 10 minutes at 4000 rpm for until a precipitate was formed. 1 mL of supernatant was taken and 0.2 M DPPH 1 mL reagent was added and 30 minutes kept in dark.A spectrophotometer at 517 nm was used to measure the absorbance.Results were then calculated using the formula and compared with those of the entire sample.Controls were measured as described above using a 0.2 M DPPH solution.The percentage antioxidant activity of soft cheese calculated with formula: % Antioxidant Activity =

Absorbance DPPH − Absorbance sample
Absorbance DPPH x 100% The data were analyzed using analysis of variance and Duncan's Multiple Range Test with a computer software SPSS version 25 for windows at a significance level of 5%.

pH, total solids, and color
The pH, color, and total solids of soft cheese added with spice powder shown in Table 1.Table 1 shows the pH, L*, a*, and b* were significantly different (P<0.05).The addition of powdered spices (cinnamon, lemongrass, and turmeric) increased the pH of cheese compared to the control (Figure 3a).Soft cheese made from 3% cinnamon powder (T3) increased the pH of the cheese from 5.60 to 6.26, resulting in an increase of 0.66, but total solids was not significantly different (P>0.05)(Figure 3b).Cheese pH can alter the size of protein aggregates and their arrangement in the protein matrix [12].Lactose mainly plays an important role in changing the pH and acidity during protein and fat breakdown, resulting in some changes in the pH of cheese [13].
The spice powder alters the initial step of milk acidification by lactic acid bacteria, thereby affecting casein aggregation in cheese.The presence of phytochemical compounds in cinnamon plants indicates that the cinnamon plant, whether dried or fresh, has medicinal potency and may contain antimicrobial, antidiarrheal, anticancer, and antihelminthic agents [14].The antibacterial compound of the spice powder can slow down the acidification and aggregation processes in cheese that can occur simultaneously.The fat and protein content of milk, particularly casein, are the primary factors affecting yield.[15].The spice powder on the total solid content of soft cheese is not significant, mainly because of the reduction in cheese yield water, spice reduced the loss of fat in the whey and water and increased the cheese yield as a result of increased fat and moisture retention in the cheese curd [16].
The average brightness value (L*) in soft cheese ranges from 66.58-80.31,redness (a*) 3.47-12.44,and yellowness (b*) 12.35-25.95(Figure 3c).The yellowness of soft cheese made from the addition of 3% turmeric powder increased from 12.35 to 25.95, resulting in an increase of 13.60.This is because turmeric has a characteristic yellow color.Turmeric also known as the golden spice because of its brilliant yellow color [17].Turmeric contains curcumin, which is yellow in color [18].Turmeric is a vibrant yellow substance that is derived from the rhizomes.Turmeric contains 5-6% of the curcuminoids [19].Compared with the control, soft cheese made with the addition of 3% lemongrass powder reduced the redness color from 5.41 to 3.47, resulting in a decrease of 1.94.In another study, consumers preferred the light-green color of lemongrass tea, with balanced bitterness and a distinct citral aroma [20].

Figure 3a
Figure 3b Figure

Antioxidant activity
Table 2 shows that soft cheese made with the addition 3% turmeric powder increased the antioxidant activity from 37.96% to 72.43%, resulting in an increase of 34.47%.Turmeric has pharmacological activities, including antioxidant, and antibacterial [21].Compared with the control, soft cheese with the addition of cinnamon powder and lemongrass powder showed increased antioxidant activity.The antioxidant activity of soft cheese made with 3% cinnamon powder increased by 22.14%.Cinnamon has biological activities, including antioxidant, antimicrobial, antifungal, and antitumor [22].The addition of 3% lemongrass powder to soft cheese increased the antioxidant activity by 28.90%.Lemongrass contains phenolic compounds that exhibit activity of antioxidant [23].
The antioxidant activity of spices is related to their chemical composition, primarily due to the biologically active compounds and presence of polyphenolic.The cinnamon contains eugenol, limonene, terpineol, catechins, tannins.Turmeric contains curcumins, essential oils, eugenol, carotene [24].The antioxidant mechanisms of protein hydrolysates may be due to the combined effects of multiple reactions that stabilize or terminate radicals, sequester pro-oxidative metal ions, and donate hydrogen [25].Milk and dairy products contain peptides that have antioxidant properties.These peptides can inhibits lipid peroxidation, chelates metal ions, and scavenges free radicals [26].In another study, cheese with fresh or dried plants had higher antioxidant activity than control.Rosemary contains rosmarinic acid exhibits the highest antioxidant activity.Rosemary cheese exhibits strong antioxidant activity [27].

Conclusion
Soft cheese with the addition of 3% cinnamon powder (T3) had the highest pH.The addition of 3% turmeric powder (T9) produced the highest level of antioxidant activity, yellowness and redness color, with the lowest brightness.In contrast, the control produced the highest brightness, with the lowest pH, antioxidant activity, and yellowness.The level addition of 3% spice powder was the most optimal level for soft cheese, with the highest pH, total solids, antioxidant activity, redness, and yellowness.

Figure 1 .
Figure 1.Process of spice powder manufacture

Figure 2 .
Figure 2. Process of spices cheese production 2.4.Determination of pH, total solids, and color A pH meter Lutron PH-220S was used to determine the cheese acidity.The total solids beginning with moisture content determination using gravimetric method, while determine the color was used a colorimeter AMT506.The colorimeter sensor is attached to the surface of the cheese and the test button is pressed.The L* (brightness), a* (redness), and b* (yellowness) values shown on the monitor screen are recorded.Color determination were carried out on 3 different surface parts of each cheese sample [10].

Figure 3 .
Figure 3. a,b,c.Graph of average pH (a), total solids (b), and color L* a* b* (c) of soft cheese with the addition of spice powder

Table 1 .
Average value pH, color, and total solids of soft cheese with the addition spice powder e Values are shown as mean ± SD.Values with different superscripts in the same column are significantly different (P<0.05).

Table 2 .
Average value antioxidant activity of soft cheese with the addition spice powder Values are shown as mean ± SD.Values with different superscripts in the same column are significantly different (P<0.05).