Retention of the Enzymatic Activity and Product Properties During Spray Drying of Pineapple Stem Extract in Presence of Maltodextrin

International Journal of Food Properties ISSN: 1094-2912 (Print) 1532-2386 (Online) Journal homepage: http://www.tandfonline.com/loi/ljfp20 Retention of the Enzymatic Activity and Product Properties During Spray Drying of Pineapple Stem Extract in Presence of Maltodextrin A.C.S. Cabral, S. Said & W.P. Oliveira To cite this article: A.C.S. Cabral, S. Said & W.P. Oliveira (2009) Retention of the Enzymatic Activity and Product Properties During Spray Drying of Pineapple Stem Extract in Presence of Maltodextrin, International Journal of Food Properties, 12:3, 536-548, DOI: 10.1080/10942910801942483 To link to this article: https://doi.org/10.1080/10942910801942483 Copyright Taylor and Francis Group, LLC Published online: 19 May 2009. Submit your article to this journal Article views: 63 View related articles Citing articles: 5 View citing articles Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalinformation?journalcode=ljfp20

International Journal of Food Properties, 12: 536 548, 2009 Copyright Taylor & Francis Group, LLC ISSN: 1094-2912 print / 1532-2386 online DOI: 10.1080/10942910801942483 RETENTION OF THE ENZYMATIC ACTIVITY AND PRODUCT PROPERTIES DURING SPRAY DRYING OF PINEAPPLE STEM EXTRACT IN PRESENCE OF MALTODEXTRIN A.C.S. Cabral, S. Said, and W.P. Oliveira Universidade de São Paulo, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Ribeirão Preto, SP, Brazil The aim of this work was to investigate the effects of drying parameters on the retention of the enzymatic activity and on the physical properties of spray-dried pineapple stem extract. A Box and Behnken experimental design was used to investigate the effects of the processing parameters on the product properties. The parameters studied were the inlet temperature of drying gas (Tgi), the feed flow rate of the pineapple extract relative to evaporative capacity of the system (W s /W max ), and the concentration of maltodextrin added to the extract (MD). Significant effects of the processing parameters on the retention of the proteolytic activity of the powdered extract were observed. High processing temperatures lead to a product with a smaller moisture content, particle size, and lower agglomerating tendency. A product with insignificant losses of the proteolytic activity (» 10%) and low moisture content (less than 6.5%) is obtained at selected conditions. Keywords: Spray drying, Pineapple extract, Proteolytic activity, Dried extract. INTRODUCTION Pineapple (Ananas comosus) is a plant widely cultivated in almost all tropical and subtropical countries. It is native to South America where wild relatives still occur. Pineapple juice has great economic interest, being one of the most important non-citrus fruit juices sold in the world. It is important also as a source of proteolytic enzymes, mainly bromelain, a group of sulfur-containing proteolytic (protein-digesting) enzymes. Bromelain is used in the food industry for meat softening and in the beer production. It has also a recognized pharmacological action, demonstrated in vitro and in vivo, as antiedematous, anti-inflammatory, antithrombotic and fibrinolytic. [1,2] The pineapple stem is one of the main wastes generated by the industrial processing of pineapple. This sub-product has high nutritional value, and may be used as a source for the extraction of bromelain and for preparation of fiber food products. [3]. In this work, it is proposed to use this material for the development of a bromelain loaded spray-dried Received 18 July 2007; accepted 25 January 2008. Address correspondence to Prof. Dr. W.P. Oliveira, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Av. do Café s/n, Bloco Q, 14040-903, Ribeirão Preto, SP, Brazil. E-mail: wpoliv@fcfrp.usp.br 536

SPRAY DRYING OF PINEAPPLE STEM EXTRACT 537 extract, which can be used as nutraceutical, flavouring agent in ice cream, sweets, confectionery, or additive in soft drink concentrates, milk-based products, and baby foods. A spray dryer is a piece of equipment wherein liquid feed is rapidly transformed into a dried particulate form by atomizing it into a hot drying environment. [4] The reduction of the degradation of thermally labile proteins and of the powder stickiness during spray drying of pineapple stem extract are typical challenges inherent to this process. The powder stickiness are mainly due to the presence of low molecular weight sugars, such as fructose, glucose, sucrose and some organic acids present in the fruit. In order to overcome these problems, drying aids such as isolated protein, maltodextrin with different dextrose equivalent (DE) are added to produce non sticky, free flowing powders. [5 8] Although the effects of adding these drying carriers on product stickiness are relatively understood, their effects on enzymes stability during spray drying is not well known. The purpose of this work was to evaluate the efficacy of the maltodextrin on the preservation of the crude enzymes stability and on physical properties of spray-dried pineapple stem extract. MATERIALS AND METHODS Materials Pineapples (Ananas Comosus), variety Perola, produced in Frutal, Minas Gerais State, Brazil, were purchased from a local market (CEASA, Ribeirão Preto SP, Brazil). Fruits were selected based on maturity, judged by colors (yellowish-greenish color). The stems were removed by hand, using a knife. The stems were pressed to squeeze out juice. The juice was filtered, added with 3 g/l of sodium benzoate, 0.5 g/l of cistein chloridrate and 2 g/l of EDTANa 2, and maintained under refrigeration (4 C) for the subsequent tests. The filtered juice was characterized by the determination of the solids, protein, reducing sugars content, and relative enzymatic activity. Spray Drying The drying was conducted in a bench-top spray dryer (model SD-05, Lab-Plant, Huddersfield, U.K), with concurrent flow regime. The drying chamber has 215 mm in diameter and height of 500 mm. The main components of the system were a feed system of the drying gas, constituted by a blower and an air filter, a temperature control system of the drying gas, and a product collect system (cyclone). The extract was fed to the spray dryer through a feed system, constituted by a peristaltic pump, a two fluid atomizer (inlet orifice diameter of 1.0 mm) and an air compressor. The drying operation started with injection of the drying air into the SD-05 spray dryer. The air was heated to the desired temperature and then the pineapple extract was fed in at a preset flow rate together with the atomizing air. Maltodextrin (MorRex 1914) kindly donated by Corn Products of Brazil was used as the drying aid, in order to evaluate its efficacy on the improvement of the stability of the crude enzymes during spray drying. Measurements of the outlet gas temperature, T go, were taken at regular intervals in order to detect the moment when the dryer attained the steady state (± 15 minutes). Once the steady state was attained, samples of the powdered product were collected. The samples were used for the physical and chemical characterization of the product, through determination of its moisture content ( volatiles content), size distribution, particle morphology, protein content, and relative enzymatic activity.

538 CABRAL, SAID, AND OLIVEIRA Physical and Chemical Characterization of the Pineapple Stem Extract The total reducing sugars in the crude pineapple stem extract was measured according to IAL (1985), [9] method 4.13.2. The total protein content was determined by the Bradford method, [10] using bovine albumin (BSA) as standard. The protein retention ratio in the spray-dried extract was defined as the ratio between the protein content in the product relative to crude extracts, as defined by Eq. (1): R TP TP,SD =, (1) T P,EX where R TP is the total protein retention ratio; T P,SD is the total protein content in the spraydried extracts; and T P,EX is the total protein content in the unprocessed stem extract (dry basis). The relative proteolytic activity of the extracts during spray drying was determined by the enzymatic hydrolysis of the casein and the quantification of the peptides formed by spectrophotometry at wavelength of 280 nm [11] (Murachi, 1976), using a spectrophotometer UV-VIS HP 8453 running the software HP Chem-Station. A unity of relative enzymatic activity was considered as the amount of the stem extract (dry basis) able to produce an increment in the absorbance reading equivalent to the release of 1 μg of tyrosine per minute under the conditions of the assay (ph 7.2 and 35 C). The reference curve was constructed with tyrosine at final concentrations ranging from 10 to 100 μg/ml. The effect of the spray drying on losses of the relative enzymatic activity was estimated through definition of the retention of the enzymatic activity ratio, R AE. This parameter was defined as the ratio between the relative enzymatic activity in the dried extract relative to crude extract (dry basis), according Eq. (2): R AE AP,SD =, (2) A where A P,SD is the relative proteolytic activity in the spray-dried extract; and A P,EX is the relative proteolytic activity of the unprocessed stem pineapple extract (dry basis). The moisture content of the spray-dried extract was determined by the oven drying method [12]. Powder samples with a pre-defined mass, m i, were placed in an oven heated at 102ºC until constant mass, m f. The percentage moisture content estimated by Eq. (3): P,EX ( m ) Up(%) = i mf 100. m f (3) The size distribution of dried product was determined by optical microscopy and image analysis. A powder sample was dispersed on the surface of a microscopic lamina. Images of the powder were obtained with the aid of an optical microscope (Olympus BX60MIV), connected to an image analysis system (Image Pro-plus 4.5). [13] The particles morphology was determined by scanning electronic microscopy (S.E.M.) in a DSM 960 Zeiss, West Germany.

SPRAY DRYING OF PINEAPPLE STEM EXTRACT 539 Experimental Design Three factors and three levels Box and Behnken design [14,15] was used to investigate the effects of the processing parameters on the product properties. The parameters studied were the inlet temperature of drying gas, T gi (70, 115, and 150ºC), the feed flow rate of the pineapple extract relative to evaporative capacity of the system, W s /W max (24, 36, and 48%) and the concentration of the maltodextrin added to the extract, MD (60, 80 and 100% relative to the solid content). These conditions were chosen based on preliminary tests. [16] The feed flow rate of the drying air was maintained constant at 0.0227 m 3 /h. The Box-Behnken design [14,15] is an independent quadratic design in that it does not contain an embedded factorial or fractional factorial design. In this design, the parameters combinations are at the midpoints of edges of the process space and at the center. These designs are rotatable (or near rotatable) and require three levels of each factor. For three factors, the Box-Behnken design offers advantages over the Box-Wilson Central Composite Designs in requiring a fewer number of runs. [17] This design allows the construction of a second-order polynomial model, which can be used to characterize or optimize a process. The resulting model has the following form: Yi = a0 + a1x1+ a2x2 + a3x3 + a4x1x2 + a5x2x3 + a6x1x3 + a7x1 2 + a8x2 2 + a9x3 2 + E, where a 0 to a 9 are the regression coefficients; X 1 to X 3 denotes the factors; Y is the relative average or expected response associate with the combination factors; and E represents the experimental error. Before the drying runs, the evaporative capacity of the spray dryer was determined using water as standard liquid material. The evaporative capacity was defined as the feed flow rate that saturates the drying gas at system outlet gas temperature. [18-19] Table 1 present the results obtained. As expected, the evaporative capacities of the SD-05 spray dryer increases with inlet gas temperature. The results of evaporative capacity were used in the definition of the feed flow rate of the pineapple extracts fed to the dryer, according the Box-Behnken design. (4) RESULTS AND DISCUSSION The crude pineapple stem has a solids concentration of 11.46 ± 0.04%, total reducing sugars of 5.05%, total protein content of 203.5 ± 7.9 μg.ml 1, and a relative enzymatic Table 1 Water evaporative capacity of the spray drying obtained by the saturation criterion. [18] T gi (ºC) T go (ºC) W max (g/min) 70 39 9.30 110 58 12.3 150 76 16.0 T gi : Inlet temperature; T go : outlet temperature; W max : evaporative capacity.

540 CABRAL, SAID, AND OLIVEIRA Table 2 Processing conditions and experimental results of the protein retention ratio, R TP, retention of the enzymatic activity ratio, R AE, and product moisture content, U P. Processing parameters Experimental results T gi ( C) W S /W max (%) MD (%) T go ( C) R TP ( ) R AE ( ) U p (%) d p (μm) 70 24 80 48 1.07 0.70 7.41 31.5 150 24 80 85 0.98 0.65 2.63 16.0 70 48 80 46 1.07 0.75 8.57 74.1 150 48 80 80 0.97 0.70 3.93 12.0 70 36 100 47 0.51 0.72 9.54 48.4 150 36 100 82 0.32 0.67 3.16 17.7 70 36 60 47 0.42 0.78 10.23 86.3 150 36 60 80 0.18 0.74 4.06 32.7 110 24 100 69 0.45 0.67 5.12 20.1 110 48 100 62 1.05 0.72 3.95 28.4 110 24 60 70 0.69 0.74 5.21 32.5 110 48 60 61 0.60 0.77 4.62 38.6 110 36 80 67 0.70 0.81 6.50 29.3 110 36 80 63 0.75 0.79 4.86 31.4 110 36 80 63 0.74 0.80 4.52 16.9 activity of 10965.2 ± 147.3 μg tyr.g 1. min 1. This extract was submitted to spray drying, according to the experimental Box Behnken design. Samples of the dried product were withdrawn during the tests, and used for their physical and chemical characterization. Table 2 presents the processing conditions and the experimental results of the protein retention ratio, R TP, retention of the enzymatic activity ratio, R AE, product moisture content, U P, and mean powder diameter, d p. Analysis of variance (ANOVA) was carried out for the experimental results presented in Table 2, in order to identify the processing parameters presenting statistical significance on total protein retention ratio, R TP ; retention of relative enzymatic activity ratio, R AE ; and product moisture content, U P. Tables 3 5 show the ANOVA analysis results, respectively for the total protein retention ratio, R TP ; retention of relative enzymatic activity ratio, R AE ; and product moisture content, U P.. ANOVA results revealed that the parameters W S /W max and MD were statistically significant at a level lower than 0.01, both for R AE and R TP. The inlet gas temperature, T gi, was highly significant for R AE and U P at a a 0.01, and only at a a level lower than 0.1 for R TP. Second order polynomial models relating the total protein retention ratio, R TP, the retention of relative enzymatic activity ratio, R AE, and the product moisture content, U p, with the processing parameters were fitted to experimental results by non-linear regression, using the least squares method. Only the parameters presenting significant effects at a a level lower than 0.05 were used in the model. Models with high determination coefficient, able to adequately describe the effects of the processing parameters on the experimental responses, were obtained (see Table 6). Figures 1a c present comparisons between the experimental results of the retention of the enzymatic activity ratio with the estimates obtained by Equation 5 (Table 6). These figures confirm the optimal agreement between the experimental data and the fitted model, exhibiting a root mean square error (RMSE) of only 0.005. As expected, these graphs show the existence of curvature effects of the processing parameters on R AE, evidencing the existence of a maximum region. This region is delimited by the maltodextrin concentrations (60 to 80%), W s /W max ratio of 36% and

SPRAY DRYING OF PINEAPPLE STEM EXTRACT 541 Table 3 ANOVA results for the total protein retention ratio, R TP. Variable Sum of squares Degrees of freedom Mean square F CALC T gi 0.05013 2 0.02507 2.952 T gi(l) 0.04805 1 0.04805 5.660*** T gi(q) 0.00208 1 0.00208 0.245 W s /W max 0.40053 2 0.20027 23.588* W s /W max (L) 0.03125 1 0.03125 3.681 W s /W max (Q) 0.36928 1 0.36928 43.496* MD 0.47328 2 0.23664 27.873* MD (L) 0.02420 1 0.02420 2.850 MD (Q) 0.44908 1 0.44908 52.895* Interaction effects T gi (L) W s /W max (L) 2.510-5 1 2.510-5 0.003 T gi (L) MD (L) 0.00063 1 0.00063 0.074 W s /W max (L) MD (L) 0.11903 1 0.11903 14.019** Error 0.04245 5 0.00849 Total 1.15360 14 *term is significant (a = 1%); **term is significant (a = 5%); ***term is significant (a = 10%); L = linear effect; Q = quadratic effect. Table 4 ANOVA results for the retention of the enzymatic activity ratio, R AE. Variable Sum of squares Degrees of freedom Mean square F CALC T gi 0.01329 2 0.00664 120.795* T gi (L) 0.00451 1 0.00451 82.045* T gi (Q) 0.00878 1 0.00878 159.545* W s /W max 0.01375 2 0.00687 124.983* W s /W max (L) 0.00405 1 0.00405 73.636* W s /W max (Q) 0.00970 1 0.00970 176.329* MD 0.00990 2 0.00495 89.956* MD (L) 0.00781 1 0.00781 142.045* MD (Q) 0.00208 1 0.00208 37.867* Interaction effects T gi (L) W s /W max (L) 0.00000 1 0.00000 0.000 T gi (L) MD (L) 0.00003 1 0.00003 0.455 W s /W max (L) MD (L) 0.00010 1 0.00010 1.818 Error 0.00028 5 0.00006 Total 0.03496 14 *term is significant (a = 1%); L = linear effect; Q = quadratic effect. temperature of 110ºC. The enzyme denaturation during spray drying of the pineapple stem extract has a complex functional relationship with the maltodextrin content and spray drying parameters. In general, the loss of the solvatation water during drying takes an important role in the protein degradation. [20] Therefore, the formation of hydrogen bonds between the OH groups of the maltodextrin (MD) and the polar groups of the protein may contribute to maintain the protein conformation (stability) during drying. The effects of the W S /W max ratio and concentration of the maltodextrin added to the extract, MD, on total protein retention ratio is presented in Fig. 2. The RMSE between

542 CABRAL, SAID, AND OLIVEIRA Table 5 ANOVA results for the product moisture content, U p. Variable Sum of squares Degrees of freedom Mean square F CALC T gi 65.49445 2 32.74723 28.335* T gi (L) 60.33511 1 60.33511 52.205* T gi (Q) 5.15934 1 5.15934 4.464*** W s /W max 2.66913 2 1.33456 1.155 W s /W max (L) 0.06125 1 0.06125 0.053 W s /W max (Q) 2.60788 1 2.60788 2.256 MD 0.96365 2 0.48183 0.417 MD (L) 0.69031 1 0.69031 0.597 MD (Q) 0.27334 1 0.27334 0.237 Interaction effects T gi (L) W s /W max (L) 0.00490 1 0.00490 0.004 T gi (L) MD (L) 0.01103 1 0.01103 0.010 W s /W max (L) MD (L) 0.08410 1 0.08410 0.073 Error 5.77864 5 1.15573 Total 75.57549 14 *term is significant (a = 1%); ***term is significant (a = 10%); L = linear effect; Q = quadratic effect. Table 6 Models fitted to the experimental results of R AE, R TP, and U p (dry basis). Fitted models R 2 2 WS WS RAE a + a1 Tgi + a Tgi + a + a a3 MD a W W + + 33 MD 0 = 0 11 2 22 2 max max 3 5 1 11 2 22 a = 0. 287; a = 6. 1 10 ; a = 3. 0 10 a = 0. 028; a = 356. 10 ; 3 5 3 33 a = 79. 10 ; a = 59. 10 4 2 (5) 0.989 WS WS W RTP = a + a + a a MD a MD a W W + + 2 0 2 22 3 33 + 23 W max max 2 3 22 3 33 S max a = 0. 310; a = 0. 211; a = 2. 21 10 ; a = 0. 116; a = 8. 710 ; 0 2 a 23 = 72. 10 4 2 P 0 1 gi 11 gi U = a + a T + a T a = 21.; 7 a = 0. 237; a = 7. 64 10 0 1 11 (7) 4 4 MD (6) 0.920 0.872 the experimental and calculated values of R TP was 0.08, confirming the adequacy of the Equation 6 (Table 6) to estimate the experimental results of total protein retention ratio obtained in this work. It can be observed that the maximum of total protein retention ratio is observed at maltodextrin concentrations of 80%, for the W s /W max ratio of 36% and T gi of 110ºC, defined previously. Moisture content (U P ) is an important property of dried products, being an indicator of the drying efficiency. In general, the reduction in the product moisture content (and water activity as well) increases the product stability and shelf life. As can be observed in Figure 3, the product moisture content is inversely related to the inlet temperature of the

SPRAY DRYING OF PINEAPPLE STEM EXTRACT 543 Figure 1 Comparison between the experimental results of R AE with estimates obtained by Equation 5 (a: MD = 60%; b: MD = 80%; c: MD = 100%).

544 CABRAL, SAID, AND OLIVEIRA Figure 2 Comparison between the experimental results of R TP with estimates obtained by Equation 6. 10 Experimental data Equation 7 U p (%kg/kg ) 8 6 4 70 110 150 T gi ( C) Figure 3 Moisture content of the dried extract of pineapple stem as a function of the outlet gas temperature. drying gas. The dotted line in Fig. 3 correspond to the fitting of the experimental results of U P with T gi, given by Equation 7 (Table 6). For the spray-dried extract obtained at optimized conditions Equation 7 estimated a moisture content around 5.0%, which agrees with the experimental values. The product size was determined by optical microscopy and image analysis. In general, the powdered extract presented a wide size distribution, with mean diameters varying from 12 to 74 μm. Fig. 4a,b present results of the product size distribution obtained at four distinct operating conditions. The dashed lines in the Figs. 4a,b represent the data fitting by Rosin-Rammler-Bennet (RRB) model. Statistical analysis performed

SPRAY DRYING OF PINEAPPLE STEM EXTRACT 545 1.0 (a) Cummulative frequency (-) 0.8 0.6 0.4 0.2 0.0 Tgi = 70 C Tgi = 150 C 0 20 40 60 80 100 120 Particle diameter (µm) 1.0 (b) Cummulative frequency (-) 0.8 0.6 0.4 0.2 Tgi = 70 C Tgi = 150 C 0.0 0 50 100 150 200 Particle diameter (µm) Figure 4 Typical results of the product size distribution as function of the inlet gas temperature, T gi : (a): Ws/ W max = 24%, MD = 80% (b): Ws/W max = 48%, MD = 80%. for the mean diameter values obtained for all experimental runs (see Table 2), indicate a significant effect of the drying gas temperature. Higher relative humidity conditions, which are often related to low drying temperatures, result in higher product moisture content during drying. Since the product has high sugar content, the increase in the moisture content, can cause the dissolution of small proportion of the sugar present, increasing the agglomeration tendency (stickiness) of the product. Figure 5 shows typical photomicrographs obtained by scanning electronic microscopy (S.E.M.), obtained at W s /W max ratio of 36%. These figures confirm that low drying gas temperatures lead to formation of agglomerated products. This behavior can be partly explained by the increase in the product moisture content, as discussed beforehand. Particles nearly spherical were obtained at temperatures of 110 and 150 C. The interaction between the maltodextrin and the extract resulted in a system of smooth (Fig. 5c), wrinkled (Fig. 5a) and dimpled (Fig. 5d) microparticles. It has been reported that sucrose can provides the formation of smooth particles when spray dried with maltodextrin and

546 CABRAL, SAID, AND OLIVEIRA a) b) 20 kv Edge = 18µm 20 kv Edge = 90µm c) d) 20 kv Edge = 18µm 20 kv Edge = 18µm Figure 5 SEM: a) 150ºC, 36% Flow rate and 100% MD, b) 70ºC, 36% Flow rate and 60% MD, c) 150ºC, 36% Flow rate and 60% MD, d) 110ºC, 36% Flow rate and 80% MD. protein. The pineapple stem extract has about 50% of sucrose and probably an favorable ratio maltodextrin: sucrose and adequate drying temperature was reached in the processing conditions used in Fig. 5c. [21] CONCLUSION The spray drying conditions, and the proportion of maltodextrin, have significant impact on preservation of the enzymatic activity, as well as on the physical properties of the spray-dried extracts of the pineapple stem. These results confirm the protective effect of maltodextrin against protein denaturation. Furthermore, as discussed by several authors, [5,6,22 25] this drying carrier improves the dryer s performance during drying of rich sugar products; as the pineapple stem extract studied in this work. The statistical analysis performed shows that the processing parameters have significant effects on product quality. The results indicate the maltodextrin concentration of 80%, the W S /W max ratio of 36% and T gi of 110ºC, as the optimized conditions for production of spray-dried extract of pineapple stem. The relative enzymatic activity of the spray-dried product obtained under the optimized conditions was at least 80% of the initial activity.

ACKNOWLEDGMENTS SPRAY DRYING OF PINEAPPLE STEM EXTRACT 547 We gratefully acknowledge The State of São Paulo Research Foundation (FAPESP), The National Council for Scientific and Technological Development (CNPq), and The Coordination for the Improvement of Higher Education Personnel (CAPES) for financial support. REFERENCES 1. Maurer, H.R. Bromelain: biochemistry, pharmacology and medical use. Cellular and Molecular Life Sciences 2001, 58, 1234 1245. 2. Kelly G.S. Bromelain: A Literature review and discussion of its therapeutics applications. Alt. Med. Rev 1996, l4, 243 257. 3. Baldini, V.L.S.; Iaderoza, M.; Ferreira, E.A.H.; Sales, A.M.; Draetta, I.S.; Giacomelli, E.J. Ocorrência da Bromelina em espécies e cultivares de abacaxizeiro. Colet. ITAL, Campinas 1993, 23, 44 55 (In Portuguese). 4. Masters, K. Spray Drying Handbook, George Godwin, Ltd., London, 3 rd ed.. 1985. 5. Bhandari, B.R.; Senoussi, A.; Dumoulin, E.D.; Lebert, A. Spray drying of concentrated fruit juices. Drying Technology 1993, 11, 1081 1092. 6. Bhandari, B.R.; Datta, N.; Howes, T. Problems associated with spray drying of sugar-rich foods. Drying Technology 1997, 15, 671 684. 7. Abadio, F.D.B.; Domingues, A.M.; Borges, S.V.; Oliveira, V.M. Physical properties of powdered pineapple (Ananas comosus) juice effect of malt dextrin concentration and atomization speed, Journal of Food Engineering 2004, 64, 285 287. 8. Jaya, S., Das, H., Mani, S. Optimization of Maltodextrin and Tricalcium Phosphate for Producing Vacuum Dried Mango Powder. International Journal of Food Properties 2006, 9, 13 24. 9. IAL. Normas analíticas do Instituto Adolfo Lutz Métodos químicos e físicos para análise de alimentos, 3 ed, São Paulo, SP, Brazil, 1985; 533 pp. (in Portuguese). 10. Bradford, M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye-binding. Analytical Biochemistry 1976, 72, 248 254. 11. Murachi, T. Bromelain enzymes. In: Lorand, L. Methods in enzymology, New York: Academic Press. 1976, XLV, 475 485. 12. WHO. Quality control methods for medicinal plant materials, Geneve, Switzerland, 1998. 13. IMAGE PRO-PLUS Reference Guide, Medya Cybernetics, Georgia EUA, 1999; 506 pp. 14. Box, G.E.P.; Behnken, D.W. Some a new three level designs for the study of quantitative variables. Technometrics 1960, 2, 455 475. 15. Box, G.E.P.; Hunter, W.G.; Hunter, J.S. Statistics for experimenters: and introduction to design, data analysis and model building; John Wiley: New York, 1978. 16. Cabral, A.C.S.; Said, S.; Oliveira, W.P. Degradação da bromelina durante secagem do extrato bruto da polpa e talo do abacaxi (Ananas comosus L. Merr.), Anais do XXXI Congresso Brasileiro de Sistemas Particulados, Uberlândia, MG, Brasil, 2004. (CD-Rom In Portuguese). 17. NIST/SEMATECH. e-handbook Statistical Methods, 2004, http://www.itl.nist.gov/div898/ handbook/ (accessed October 12, 2005). 18. Nath, S.; Satpathy, R.A Systematic approach for investigation of spray drying process. Drying Technology 1998, 16, 1173 1193. 19. Souza, C.R.F.; Oliveira, W.P. Powder properties and system behavior during Spray drying of Bauhinia forficata Link extract. Drying Technology. 2006, 24, 735 749. 20. Sonner, C. Protein-loaded powders by spray freeze drying. Thesis, Ph.D., Friedrich-Alexander Universitat, Germany, 2002; 151 pp. 21. Tzannis, S.T.; Prestrelski, S.J. Moisture effects on protein-excipient interactions in spray dried powders. Nature of destabilizing effects of sucrose. J. Pharm. Sci. 1998, 88, 360 370. 22. Adhikari, B.; Howes, T.; Bhandari, B.R.; Truong, V. Characterization of the surface stickiness of fructose-maltodextrin solutions during drying. Drying Technology 2003, 21, 17 34.

548 CABRAL, SAID, AND OLIVEIRA 23. Adhikari, B.; Howes, T.; Bhandari, B.R.; Truong, V. Effect of addition of maltodextrin on drying kinetics and stickiness of sugar and acid-rich foods during convective drying: experiments and modeling. Journal of Food Engineering 2004, 62, 53 68. 24. Bhandari, B.R.; Datta, N.; Crooks, R.; Howes, T.; Rigby, S. A semi-empirical approach to optimize the quantity of drying aids required to spray dry sugar rich foods. Drying Technology 1997, 15, 2509 2525. 25. Mani, S.; Jaya, S.; Das, H. Sticky issues on spray drying of fruit juices, 2002 ASAE/CSAE North-Central Intersectional Meeting, Saskatoon, CANADA, September 27 28, 2002, paper no: MBSK 02 201, ASAE, 2950 Niles Road, St. Joseph, MI, USA.