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Development and Optimization of Instant Drink Formulation from Pineapple (Ananas cosmosus) Juice

Steffi1, Maruli Pandjaitan2 and Hery Sutanto1

1Department of Pharmaceutical Engineering, Swiss German University, Serpong, Indonesia 2Department of Biomedical Engineering, Swiss German University, Serpong, Indonesia

(Tel: +62 21 − 3045 0045; E-mail: [email protected])

Abstract – Pineapple (Ananas cosmosus) has been proven to have many beneficial effects for health due to its bromelain content. The formulation of an instant drink form fresh pineapple juice to produce an instant drink product with high bromelain content was developed. A number of pineapple powder specimens were produced using a spray dryer with different formulations. Fresh pineapple juices were added with 15% of maltodextrin (MD) combined with 0%, 0.1%, 0.2%, 0.3%, 0.4%, and 0.5% of carboxymethyl cellulose (CMC) before exposing to the drying temperature at 150°C. The qualities of pineapple powders were investigated in the aspects of enzyme activity, moisture content, solubility, and vitamin C content. The results indicated that the preferred formulation for spray dried pineapple was the addition of MD at 15% and CMC at 0% with specific activity of 2.212 ± 0.459 U/mg, moisture content of 4.27 ± 0.177 %, solubility of 0.83 ± 0.236 g/ml, and vitamin C content of 0.466 ± 0.029 mg/g pineapple. Keywords: Pineapple, Bromelain, Spray Drying, Maltodextrin, Carboxymethyl Cellulose

I. INTRODUCTION

Pineapple or Ananas cosmosus is a tropical fruit belongings to the Bromeliaceae family. Pineapple has several health benefits due to its rich content of various minerals (manganese, phosphorus, iron, calcium, magnesium, and potassium) and vitamins such as vitamin B1, B2, B3, B6, and C. High vitamin C content in pineapple cause the pineapple to possess antioxidant properties that promote the elimination of free radicals from the body [1]. Therefore, pineapple consumption gives beneficial effect for human health, especially for the immune system due to its ability to eliminate pathogens.

Pineapple is not only used as a food source, but also as traditional medicine because the parts of pineapple can be useful for treating and relieving illness due to its proteolytic enzyme called bromelain. Recent studies have discovered that bromelain can be effectively used to treat HIV, Hepatitis C, and cancer patients. The anti-cancer property of bromelain is due to its protease components [2].

Bromelain is a protease enzyme that is found mostly in the pineapple stem. Bromelain can be absorbed in human intestines without degradation and losing its biological

activity [3]. Its function is to break the peptide bonds of proteins and cleave the protein into amino acid. Bromelain can be used to treat wounds, burns, infections, sprains, and swelling and inflammation after sinus surgery [4].

Bromelain is effective to remove mucus, therefore it is often used in sinus or asthma treatment. It stimulates the production of cytokine, which is very useful to treat the cancer patients because cytokines are produced by leukocytes which increase the immune system and fight the pathogens [5]. Since it has various therapeutic effects as anti-inflammatory, anti-cancer, anti-thrombotic, and fibrinolytic agents, bromelain is very valuable in the medicinal sector.

Various instant products from pineapple can be produced by utilizing the current technology, such as instant pineapple juice drink production by using a spray drying technique. In this research, the formulation of instant drink from pineapple juice with a high concentration of bromelain will be developed and optimized. Pineapple in powder form is beneficial because it has a long shelf life at ambient temperature, is convenience to use, and has low transportation expenditure. An instant drink product of pineapple juice which contains bromelain concentration will give benefits to the consumers due to its potent and practical usage.

In this research, spray drying method was used to produce instant drink product from pineapple juice. The fillers used in this research were maltodextrin (MD) and carboxymethyl cellulose (CMC).

II. METHODS

Raw materials used for the preliminary research were ripe

pineapples from Bogor and Palembang. The pineapple fruit was selected based on color and the outer appearance of the fruits (no bruises). The pineapples were purchased from a supermarket in BSD City, Tangerang.

The pineapple fruit was peeled, its eyes were removed and cut into small pieces. The fruit was added cold distilled water with 1:1 ratio of weight to volume, 1 ml of distilled water was added to every 1 g of fruit. The fruit was blended at speed setting 1 for 5 minutes. The juice was filtered through a filter cloth and analytical sieve of 36 μm and 20 μm.

2013 3rd International Conference on Instrumentation, Communications, Information Technology, and Biomedical Engineering (ICICI-BME)

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Bandung, November 7-8, 2013

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Each 300 ml of filtered pineapple juice was added with 15% of MD and variation concentration of CMC at 0%, 0.1%, 0.2%, 0.3%, 0.4%, and 0.5%. The spray drying process was performed at temperature of 150°C at the inlet. The final product was analyzed in the aspects of enzyme activity, moisture content, solubility, and vitamin C content.

Enzyme Activity Analysis 125 μl sample, 125 μl calcium chloride (CaCl2) 0.04 mM, 250 μl casein 1%, and 250 μl phosphate buffer pH 7 were added into a microtube and vortex. The microtube was incubated with 37°C water for 20 minutes. 750 μl of 5% trichloroacetic acid (TCA) was added into the microtube and vortex. The microtube was centrifuged at 4°C and 8000 rpm for 10 minutes. 300 μl sample from the microtube was transfered into a test tube. 1000 μl sodium carbonate (Na2CO3) and 200 μl Folin-ciocalteau (1:2) were added into the test tube and vortex. The sample was poured into a cuvette and its absorbance was measured in a spectrophotometer at wavelength 578 nm. For the blank solution and standard solution, the same procedure was applied. Tyrosine with different amounts of concentration was used as the standard solution. Lowry method was used to determine the protein content. The specific activity of the enzyme was calculated by dividing the volume activity with the protein content.

Moisture Content Analysis Moisture content analysis was performed by using Sartorius MA35 moisture content analyzer. The temperature was set to 105°C and the time was set to 30 minutes. After the moisture content analysis was completed, the percentage of moisture content was shown on the display screen. The water content of the sample was analyzed in w/w percentage.

Solubility Analysis Solubility analysis was performed by using micropipette, beaker glass, and glass rod. The solubility of the sample was determined in g/ml.

Vitamin C Content Analysis A redox titration using iodine was applied to analyze the content of vitamin C in sample. 10 ml sample, 100 ml distilled water, 50 ml sulfuric acid (H2SO4) 1 N, and 1 ml starch 1% were added into an Erlenmeyer flask. The solution was titrated with iodine 0.01 N until the solution became blue and remained. The titration was repeated three times. The mass of ascorbic acid was calculated for each sample.

The experiment data were compared using Analysis of Variance (ANOVA) Single Factor and t-test analysis.

III. RESULT AND DISCUSSION

Preliminary Research Two types of ripe pineapple were

used for the preliminary research, namely Bogor pineapple and Palembang pineapple. Both types of pineapple were evaluated for their volume activity, protein content, and specific activity. The result of specific activity comparison is shown in Fig. 1.

Fig. 1. Specific activity between Bogor and Palembang

pineapple

Based on the result shown in Fig. 1., the enzyme activity of Bogor pineapple is higher than Palembang pineapple. This was due to the higher absorbance that was exhibited during reading in the spectrophotometer. It means that the enzyme activity in Bogor pineapple was higher because it hydrolyzes more proteins from casein into single amino acids tyrosine. This resulted in more amino acids obtained in the cuvette and yielded high absorbance.

The statistic result from t-test also showed that the enzyme activity analysis between Bogor pineapple and Palembang pineapple had significant difference (p<0.05). Therefore, Bogor pineapple was selected to be used for further research.

Primary Research The filtered pineapple juices made from Bogor pineapple were added with 15% MD and various concentrations of CMC and spray dried. The reconstituted pineapple powders were analyzed for their enzyme activity, moisture content, solubility, and vitamin C content.

Enzyme Activity Analysis The specific activity of bromelain in pineapple powders is shown on Fig. 2.

Fig. 2. Specific activity between pineapple juice and

spray dried pineapple

Fig. 2. shows that the combination of 15% maltodextrin with 0.4% carboxymethyl cellulose yielded the highest enzyme activity. However, the result of statistical analysis by using ANOVA Single Factor stated that there was no significant difference (p>0.05) in enzyme activity under various treatment. It means that the various treatment of carboxymethyl addition did not significantly preserving the enzyme activity. The phenomenon might occur due to small concentration of carboxymethyl cellulose addition.


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Moisture Content Analysis The outcomes of moisture content for the pineapple powders are presented in Fig. 3.

Fig. 3. Effect of CMC addition on moisture content of

spray dried pineapple

Based on Fig. 3, the spray dried products had moisture content in the range of 4.27%-5.02%. Reference [6] stated that typical moisture content of powder ranges from 3-6% (w/w), therefore the moisture content of spray dried pineapple from various carboxymethyl cellulose concentrations was still acceptable. The moisture content was slightly increased by the addition of higher concentration of carboxymethyl cellulose. The carboxymethyl cellulose acted as water-binding material, thus it hold water and prevent moisture removal during the heating process. Therefore, the addition of carboxymethyl cellulose caused the evaporation rate became lower [7].

The spray dried products from carboxymethyl cellulose variation yield in significantly different (p<0.05) moisture content as proven by Analysis of Variance (ANOVA) Single Factor test. According to the t-test done to each data, there was a significant difference of moisture content between the product of 15% maltodextrin and 15% maltodextrin with 0.5% carboxymethyl cellulose. This also was because carboxymethyl cellulose is easy to absorb moisture, because carboxymethyl cellulose can bind water via hydrogen bond [8]. Therefore the highest moisture content was in the product with the highest concentration of carboxymethyl cellulose, which was 0.5% of carboxymethyl cellulose.

Solubility Analysis The result of solubility analysis of spray dried pineapple is shown on Fig. 4.

Fig. 4. Effect of CMC addition on solubility of spray dried pineapple

Based on Fig. 4., the addition of carboxymethyl cellulose,

in all concentrations, decreased the solubility of the product. Reference [9] stated that the addition of carboxymethyl cellulose negatively affected the solubility of spray dried powder. The reconstitution property from the pure extract is better than those prepared by addition of carboxymethyl cellulose.

The result of ANOVA Single Factor test shows that the addition of carboxymethyl cellulose gave no significant difference (p>0.05) to solubility of all spray dried products.

Vitamin C Content Analysis Prior to spray drying process, the vitamin C content measured in pineapple juice was 0.837 ± 0.014 mg/g pineapple. The result vitamin C content after fillers addition and spray drying process was shown on Fig. 5.

Fig. 5. Vitamin C content of pineapple juice and spray

dried pineapple From the result, the vitamin C content in the sample

product was decreased compared to the vitamin C content before spray drying. The heating exposure during spray drying might cause the decrease in vitamin C content as vitamin C is very relatively unstable to heat. Vitamin C is very susceptible to destruction by oxidation [10]. The process is accelerated by the presence of heat, light, alkali, enzyme, oxidator, and catalyst of copper and iron.

However, the bar graph from each sample from Fig. 5. were fluctuating and the result of ANOVA test showed no significant difference (p>0.05) in carboxymethyl cellulose addition on vitamin C content in spray dried pineapple. This result showed that various additions of additives did not affect the vitamin C content in pineapple powders.

Reference [11] also reported that the addition of carboxymethyl cellulose gave no significant effect towards the vitamin C content in pineapple jam. The result showed the fluctuation of vitamin C content as the concentration of carboxymethyl cellulose was increased. The heating treatment that was required to process the pineapple caused the decrease in vitamin C content in the pineapple jam if compared to the vitamin C content in the original pineapple fruit.


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Thus, in this research, the quality of vitamin C was reduced after spray drying because of high temperature and oxidation. On the other hand, the addition of carboxymethyl cellulose did not affect the vitamin C content as it did not give significant difference towards vitamin C content.

IV. CONCLUSION

From the analysis conducted, it can be concluded that the

addition of carboxymethyl cellulose did not significantly affect the enzyme activity, vitamin C content, and solubility of spray dried pineapple. However, the result of moisture content analysis showed significant difference among formulation with carboxymethyl cellulose variation. The more carboxymethyl cellulose added into the formulation resulted in higher moisture content of the spray dried pineapple. Since lower moisture content is preferred for powder product, thus the formulation with 15% maltodextrin was selected for spray dried pineapple. Therefore, the preferred formulation for spray dried pineapple was the addition of 15% of maltodextrin with specific activity of 2.212 ± 0.459 U/mg, moisture content of 4.27 ± 0.177 %, solubility of 0.83 ± 0.236 g/ml, and vitamin C content of 0.466 ± 0.029 mg/g pineapple.

Actually the addition of maltodextrin and carboxymethyl cellulose could preserve the enzyme activity of spray dried pineapple. However, the statistical analysis resulted in insignificant difference between carboxymethyl cellulose variations. Furthermore, the result of enzyme activity of spray dried pineapple before (pineapple juice) and after (spray dried pineapple with 15% of maltodextrin) spray drying was not different significantly. Therefore, the selected formulation for spray dried pineapple was 15% of maltodextrin.

REFERENCES

[1] Joy, P.P. 2010. Benefits and uses of pineapple.

http://www.kau.edu/prsvkm/Docs/Benefitsandusesofpineapple.pdf, accessed on January 09, 2013.

[2] Chobotova, K., A.B. Vernallis, and F.A.A. Majid. 2010. Bromelain’s activity and potential as an anti-cancer agent: Current evidence and perspectives. Cancer Letters 290: 148-156.

[3] Castell, J.V., G. Friedrich, C.S. Kuhn, and G.E. Poppe. 1997. Intestinal absorption of undegraded proteins in men: presence of bromelain in plasma after oral intake. Am. J. Physiol 273 (1): 139-146.

[4] Maurer, H.R. 2001. Bromelain: biochemistry, pharmacology and medical use. Cell Mol Life Sci. 58(9): 1234-1245.

[5] Kelly, G.S. 1996. Bromelain: a literature review and discussion of its therapeutic applications. Alt Med Rev 1 (4): 243-257.

[6] Johnson, R. 2010. Pharmaceutical spray drying. http://www.slideshare.net/xtalks/pharmaceutical-spray-drying-upperton, accessed on June 20, 2013.

[7] Alam, K., M. Ahmed, S. Akter, N. Islam, and J. Eun. 2009. Effect of carboxymethylcellulose and starch as

thickening agents on the quality of tomato ketchup. Pakistan Journal of Nutrition 8(8): 1144-1149.

[8] Boruvkova, K., J. Wiener, and M. Jakubickova. 2012. Preparation and properties of microporous structures based on CMC. http://www.nanocon.eu/files/proceedings/04/reports/758.pdf, accessed on June 22, 2013.

[9] Abdalla, A.A., M.A. Mohammed, and H.A. Mudawi. 2010. Production and quality assessment of instant boobab (Adansonia digitata L.). Advance Journal of Food Science and Technology 2 (2): 125-133.

[10] Winarno, F.G. 2008. Kimia Pangan dan Gizi. Bogor, Indonesia: M-Brio Press.

[11] Syahrumsyah, H., W. Murdianto, and N. Pramanti. 2010. Effect of carboxy methyl cellulose (CMC) addition and maturity level of pineapple (Ananas cosmosus (L) Merr.) on quality of pineapple jam produced. Jurnal Teknologi Pertanian 6(1): 34-40.


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