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INDONESIAN SCHOLARS JOURNAL Paper Number (Will be replaced with volume and paper number if the manuscript is accepted)

Browning Intensity, Antioxidant Activity, and Goaty Aroma of Goats Whey Glycated by Lpsicose, L-tagatose, and L-fructose
1

Y.A.P. Waskito* & 2F.G.Salim

Faculty of Animal and Agriculture Sciences, Diponegoro University 2 Faculty of Engineering, Diponegoro University

Abstract. Rare Sugar added to whey is predicted to be usefull for reducing the risk of obesity and increasing its functional property.The purpose of this research was determine the browning intensity, antioxidant activity, and goaty aroma of goats whey that was glycated with 4% L-psicose, L-tagatose, and L-fructose at 65 C for 30 minutes. Browning intensity was measured by spectrophotometry at 420 nm, antioxidant activity was determined by spectrophotometry mean of ABTS as substrate and goaty aroma was determined by organoleptic with 25 respondents. The design of an experiment to test the browning intensity, antioxidant activity, and goaty aroma was completely randomized design (CRD) with 3 treatments and 7 replications, if there is a significant effect of treatment followed by Duncan Multiple Test Area. The treatments was the addition of rare sugar as much as 4% L-psicose (T1), 4% L-tagatose (T2), and 4% L-fructose (T3). The results showed that the addition of rare sugar (4%) gave a significant influence (P>0.05) to browning intensity, antioxidant activity, and goaty aroma. Browning intensity generated a number from 0.186-0.611%; antioxidant activity 14.403-37.610%; and goaty aroma 3.88-5.08. The conclusion of this research was goats whey with rare sugar (L-psicose, L-tagatose, and L-fructose) increased functional properties such as browning intensity, antioxidant activity, and decrease goaty aroma in goats whey.

Keywords: antioxidant, browning intensity, goaty aroma, rare sugar, whey

A. INTRODUCTION
Whey is the liquid remaining after milk has been processed through coagulation and filtration. It is a major by-product of cheese or casein manufacturing, which has versatility with respect to functionality, and high nutritive value as an ingredient in food products. Whey-based food products has yet to be commonly found in the society, due to peoples tendency to choose sweet food products. While sugar added foods are popular, adding conventional sugar to whey may increase the risk of obesity. For that reason, rare sugar is expected to become the substitute for conventional sugar. Rare sugar is the derivative product from starch or lactose, that has low, or almost-zero calories. Since it also contains a high level of antioxidant activity, it may potentially be used for daily consumption. Rare sugar is very different than polyol or alcoholic sugar, for polyol has the molecular formula H(HCHO)n+1H, while rare sugar has the same molecular formula as monosaccharide, (H(HCHO)nHCO). Because of the similarity in the molecular formula, it is expected that rare sugar and conventional sugar have similar properties (for example, the ability to cause Maillard effect). This would allow rare sugar to substitute sucrose for industries. especially for those who want to have the Maillard effect on their products, such as pudding and cake industries [1]. According to the International Society of Rare Sugar (ISRS), there are 34 types of rare sugar, the amount of research conducted internationally for rare sugar application has been increasing significantly. This increase is triggered by the growing popularity of healthy lifestyle, as well as the unique properties of rare sugar, which have motivated many food researchers in the world to research rare sugar as an additive for functional food, so that it may benefit the society [1] further. The health benefits of rare sugar, among others, is what makes rare sugar a rather unique research topic. In field of food, D-psicose, for example, is a zero-calorie sugar and has been proven to have a potential to be a natural antioxindant, as reported in many food research. D-piscose is used for sucrose substitution in cake. Subtituting 20% Dpsicose in a cake helps create better texture, browning, and it also boosts the color. D-psicose is reported to have a low glycemic index, or in other words, it does not increase ones blood sugar concentration, which helps prevent obesity and diabetes [1]. Glycation is a chemical reaction that happens when sugar molecules are heated to a certain temperature, bonds with a protein molecule, and produces a new compound called melanoidin. Melanoidin compound has many benefits, especially for the food industry, such as increasing browning

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intensity, increasing antioxidant activity, and reducing goaty aroma in food products manufactured from goat milk. As of now, in Indonesia, the research on the application of rare sugar in food product such as whey goat milk has not existed. The combination of the unique properties of rare sugar that has many benefits, and that of whey goat milk, which are also equally unique--such as distinctive odor from low chain fatty acid, enzyme or amino acid essentiality which is abundantly found in whey, and its function as an antioxidant, is an interesting topic to be researched further, as it may provide guidance to food industry companies on the best way to treat whey as a by-product of cheese and casein manufacturing. The purpose of this research is to analyze shifting of browning intensity, antioxidant properties, and goaty aroma caused by glycation process of whey goat milk with adding Lpsicoes, L-tagatose, dan L-fructose. The benefits of this research is to know shifting browning intensity, antioxidant properties, dan goaty aroma of goats whey glycated by L psicose, L-tagatose, and L-Fructose sugar in particular precentage

B.4 Determination of Antioxidant activity Pour 0,4 gram of L-psicose, L-tagatosa and L-fructose into microtube and each microtube is added 1ml whey goat milk and vortex them. That sample is heated at 65 oC for 30 min and then leave it for 10 min. 300L dan 2700Lis taken for antioxidant test kit and cover microtube and test kit with alumunioum foil and incubated for 2 hourd. After that, open alumunium foil and vortex it. pick 3mL sample to cuvvete then see the absorbance value to know antioxidant value for 734 nm. Absorban is expected on 0,2 -0,6 range [3] . B.5 Determination of Browning Intensity Determinatuion of browning intensity using fosfat buffer with pH at 7,4 and spectrophotometer at 420nm. Previously, 7,8 gram sodium dihydrogen fosfat (pH 4) is dissolved into 500 ml aquades and 17,907 gram of sodium hydrogen fosfat (pH 9) is dissolved into 500 ml aquades. 100 ml of sodium dihydrogen fosfat is taken and add sodium hydrogen fosfat gradually . check pH and stop when its pH 7,4. 30 l taken from sample is poured into cuvvet and add 2970 l buffer fosfat solution . Sample is tested with spectrophotometer with 420 nm wave in order to know the absorbance value [3]. B.6 Determination of goaty aroma Pour 0,4 gram of L-psicose, L-tagatosa and L-fructose into microtube and each microtube is added 1ml whey goat milk on each tube and each sample is heated in waterbath during 30 min at 65oC. sample is cooled during 10 min. just about right with 9 criteria is used in organoleptic test. Organoleptic test is tested to 25 correpondents [4].

B. METHODS
B.1 Materials Reasearch about Shifting of browning intensity, Antioxidant properties, and Goaty aroma of goats whey glycated by L-psicose, L-tagatose, and L-fructose held on October 2012 till Februari 2013 in Chemistry and Nutrition food labroratory, Faculty Animal and Agriculuture Sciences Diponegoro university. All chemicals used is this study were of analytical grade and All were obtained from Chemistry and Nutrition food labroratory B.2 Preparation of Whey goat milk

C. RESULTS
Adding 4% rare sugar into whey goat milk showed a distinctive result (P>0,05) against browning intensity, and goaty aroma. Analysis of browning intensity, antioxidant activity, and goaty aroma with various sugar can be seen in figure 1

1 liter goat milk was adjusted at 27 C. then prepare rennet that was diluted into 50 ml aquadest. Adjust waterbath at 35oC then wait till milk temperature is same with waterbath temperature. Pour lactat acid to milk till its pH at 6.0. Pour rennet and mix it then leave it for 40 min. milk will be coagulated and be diced, leave it for ten min then filter it to obtain whey. And then keep whey into freezer , keep curd (the solid one) inside closed container in refrigerator [2]. B.3 Preparation of ABTS (2,2-azinobis(3ethylbenzothiazoline-6-sulphonic acid) solution Dilute 0,08 gram ABTS powder with 21 ml aquadest and prepare PP solution with diluting 0,014 gram PP with 21 ml aquadest. Two solution is poured together into centrifuge tube and its tube cover with alumunium foil in order to avoid it from light. Mix them until that is homogenous [3].

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to brownish. Color is one of indicators that chemical reaction is happened on food. [6] C.2. Antioxidant activity of whey goat milk with adding various type of sugar Fig.1 showed that antioxidant activity value for T1 (intercalation 4% L-tagatosa) is 14,40% , T2 (intercalation 4% L-psikosa) is 37,61 %, and T3 (intercalation 4% L-fruktosa) is 25,22%. Based in various antioxidant activity analysis showed that adding various sugar on whey in glycation process influenced antioxidant activity. It is proven that actual F theoretical F so that H0 rejected and H1 accepted. Further analysis using Duncan double area showed that T1 treatment is obviously different with T2 and T3. T2 treatment is obviously different with T1 and T3. Treatment T3 is obviously different with T1 even T2. Adding sugar on whey in glucation process could increase antioxidant activity up to 14,40%-37,61%. An existing of antioxidant can be detected by ABTS method. This compound is obtained from oxidation of calium persulfat with diammonium salts and ABTS. An existing antioxidant activity from sample was causing a shifting color on ABTS solution originally blue greeness shifting to colorless. Shifting color indicated that free radical of ABTS captured by antioxidant compound which was contained in sample. Percentage inhibition in this report is deviation of absorbance between absorbance control ( with no sugar added) with absorbance sample. Lingga [7] reported that antioxidant is a natural compound that keep our cell body from damage caused by reactive molecule or called free radical. It obstructs oxidative reaction with binding free molecule and keep the genetic structure of cell in order to keep in normal condition. The compound that contain particular bioactive which has antioxidative ability can weaken another compound that reactive if it reacts with oxygen. Oxidative reaction can be obstructed with reduction. Furthermore antioxidant is called reductive compound The increase of antioxidant in adding of sugar is happened during heating when the reaction between sugar and whey goat milk produced a compound that has antioxidant properties. That compound has an influence for the increase of antioxidant. Nagai et al [8] reported that glication is a reaction happened between amino group and carbonil grup, perticulary reduction sugar. The continuous glycation will form melanoidin such as end product from maillard process [9]. Maillard reaction formed during heating and long incubation can produce antioxidant in significant amount. Antioxidant

Fig.1. Mean value of browning intensity, antioxidant activity, and goaty aroma on whey goat milk in glycation process with adding various type of sugar (4%)

C.1 Browning Intensity with adding various type of sugar Glycation process of whey goat milk with adding 4% Ltagatosa (T1) showed absobance value 0,5390,019 b, adding 4% L-psikosa (T2) 0,6110,074 and adding 4% L-fruktosa (T3) 0,1860,077c (Fig 1). The higher absorbace value, the more brownish is resulted. Adding various sugar on whey in glycation process showed an obvious influence (P<0,05) againts browning intensity. This is proven that actual F theoretical F so that H0 rejected and H1 accpeted. Further analysis using Duncan area showed that T1 treatment is very different with T2 and T3. T2 treatment is very different with T1 and T3. T3 treatment is very different with T1 and T2. T2 treatment (intercalation L-psikosa) showed highest asorbance value. The asorbance value of T1 treatment ( intercalation Ltagatosa) is higher than T3 treatment (intecalation L-fructose) and asorbance value of T3 is the lowest The increase of absorbance value showed that shifting color of whey into brownish is happened. Brown color formed is caused by a reaction between carbonil group from sugar containing amino group on protein in whey during heating. According to reineccius [5] that maillard reaction produced much compound and could influence a color of product. A reaction between reductive sugar and amino acid through maillard reaction resulted a shifting color of food from yellow

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activity of T2 (intercalation L-psikosa) has the highest antioxidant activity. Antioxidant activity of T3 (intercalation L-fruktosa) is higher tham T1(intercalation L-tagatosa). Ltagatosa has the lowest antioxidant activity. It is caused by adifferent structure and reduction properties from another sugar types so that it influenced the ability of L-tagatosa in binding free radical compound came from ABTS. Antioxidant activity produced from various treatment, is caused by a different structure and reductive properties from each sugar. C.3 Goaty aroma against whey goat milk with adding various type of sugar Fig.1 showed that a mean value of goaty aroma against T1 treatment (intercalation 4% L-tagatosa) is 5,08 ; T2 treatment (intercalation 4% L-psikosa) is 3,88 and T3 ( intercalation 4% L-fruktosa) is 4,00. Organoleptic test score for the rate of aroma produces is between 1 to 9 with highly no smell to highly smell. The mean score in T1 treatment was in netral to goaty category with 5,08 ; T2 treatment (adding L-psikosa) was in no goaty category to goatless with 3,88 and T3 (adding L-fruktosa), was in no goatless with 4,00. According to analysis manner showed that adding various type of sugar obviously influenced against aroma. It is proven that actual F theoretical F so that H0 rejected and H1 accepted. Further analysis used obvious different showed that T1 treatment is obviously different with T3 and different with T2. While T2 treatment is obviously different with T1, but not too different with T3, then T3 is obviously different with T1 but not obviously different with T2 Whey goat milk has a distinctive goaty smell, but after whey goat milk suffered glycation process with rare sugar, goaty aroma reduced even undetected. Legowo et al [10] reprted that Prengus taste and aroma sticks to goaty milk. short chain fatty acid kaprilat and short chain fatty acid laurat is in the highest amount in goat milk and influenced against prengus taste and aroma goat milk. Heating goat milk with sugar addition, resulted a distinctive aroma. That aroma can reduce goaty aroma in goat milk. It was appropriate with Mejcher and henry [11] study that furaneol is formed in food product contained reductive sugar and amino acid. Furaneol compound (4-hidroksi-2,5-dimetil-3-(2H)- furanon) formed through maillard reaction from 2 hidroksil propanol. Aroma can be detected by sense of smell (nose). According to deMan [12], Aroma is usually caused from an existing solution from smelly compound. In maillard reaction, karbonil compound especially fufural, hidrokksimetilfurfural, and aldehid, is an important smelly compound.

C.4 Browning intensity, antioxidant activity, and goaty aroma in whey goaty milk with adding various type of sugar Whey goat milk with adding rare sugar in particular concentration (4%) showed an obviously distinction on browning intensity parameter, antioxidant activity, and goaty aroma from whey goat milk. This is because whey goat milk suffered alteration of functional properties which is obviously different after adding with various type of sugar, so that browning intensity, antioxidant activity and goaty aroma showed a significant alteration from various type of sugar addition. It was appropriate with Liu et al study [13] This is because a reaction between reductive sugar with amino acid contained in each sugars. Glycation is happened when protein in food against sugar which is heated on particular temperature so that it will produce a new compound called melanoidin compound. Melanoidin compound has a benefit especially in food industry: increasing browning intensity caused by maillard reaction, increasing antioxidant ammount, and covering goaty aroma on food product

D. CONLUSIONS
According to the experiment result can be concluded that whey goat milk with particular percentage of various rare sugars addition (4%) showed an obvious influence against browning intensity, antioxidant activity, and goaty aroma. Adding L-psikosa result the highest level of browning intensity, reducing goaty aroma, and antioxidant activity. Adding L-fruktosa resulted the high level of antioxidant activity. The higher browning intensity is resulted in glycation the higher antioxidant activity is going to be. Glycation with adding various type of sugar can increase browning intensity, reducing goaty aroma, and increasing antioxidant activity on whey goat milk

ACKNOWLEDGEMENT
The authors would like to thank Ahmad Nimatullah AlBaarri, Ph.D and Setya Budi Muhammad Abduh, M.Sc for supporting this research

REFERENCES
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pp. 509-517, 2005. [4] Z. Vickers, "Sensory Spesific Satiety in Lemonade Using Just Right Scale for Sweetness," Journal of Sensory Studies, vol. I, no. 3, pp. 1-8, 1988. [5] G. Reineccius, Flavor Chemistry and Technology 2nd Edition, LLC: Taylor and Franchis Group, 2006. [6] S. Nielsen, Food Analysis 3rd Edition, vol. III, New York: Plenum Publisher, 2003. [7] L. Lingga, The Healing Power of Antioxidant, Jakarta: PT. EleX Media Komputindo, 2012. [8] R. Nagai, M. Jinno, M. Ichihashi, H. Koyama, Y. Yamamato and Y. Yonai, "Advanced Glycation end Product and Their Receptor as Risk Facctors for Aging," Anti Aging Magazine, vol. IV, no. 9, pp. 108-113, 2012. [9] M. Nicoli, M. Anese, M. parpinel, S. Franceschi and C. Lerici, "Study on Loss and formation of Antioxidants during Processing and Storage," Cancer Letters, no. 114, pp. 71-74, 1997. [10] A. Legowo, A. Al-Baarri, M. Adnan and U. Santosa, "Identifikasi dan Karakterisasi Aroma "Prengus" ("Goaty Flavour") Susu Kambing serta Produk Olahannya," International Journal of Tropical Animal Production, no. 31, pp. 1-10, 2006. [11] M. Mejcher and H. Henryk, "Idenstification od Potent Odorants Formed during the Preparation of Extruded Potato Snacks," J.Agric.Food Chemistry, no. 53, pp. 6342-6347, 2005. [12] J. deMan, in Kimia Pangan Edisi Kedua, Bandung, Penerbit ITB, 1997. [13] J. Liu, R. Qiaomei and Y.Ding, "2012," Glycation a Promising Method for Food Protein Modification: Physicochemical Properties and Structure, a Review, no. 49, pp. 170-183, 2012.