Journal

of Controlled

Release, 7 (1988) 33-37

33

Elsevier Science Publishers

B.V., Amsterdam

Printed

in The Netherlands

AMYLOSE CONTENT BIOACTIVE AGENTS Robert E. Wing,

Northern (U.S.A.)
(Received

OF STARCH

CONTROLS

THE RELEASE OF ENCAPSULATED

Sukumar

Maiti*

and William

M. Doane
of Agriculture*,Peoria, IL 6 1604

Regional Research Center, Agricultural

Research Service, U.S. Department

June 23,

1987;

accepted

in revised

form September

15,

1987)

Controlled release of the herbicide butylate [S-ethyl bis(2-methylpropyl) carbamothioate] from a non-chemically modified starch matrix has been achieved. Steam injection cooking of cornstarch or mixtures of starches with varying amylose content, followed by active agent addition, drying and grinding yields encapsulated products showing controlled slow release of active agent. As the amylose content increases, the rate of release decreases due to the ability of amylose to retrograde (starch chain association). As the amylose content is reduced to about IO% the efficiency of active agent encapsulation nears 100%. Encapsulated products consisting only of starch and an active agent should be useful in a variety of applications, i.e. food, feed or agriculture.

INTRODUCTION

Recently we reported [ 1 ] a new method for encapsulating bioactive agents within a jetcooked pearl cornstarch matrix. Pearl cornstarch is a low-cost and abundant polymer composed of a mixture of amylose ( - 25% ) and amylopectin ( - 75% ) . Amylose is essentially a linear polymer having a molecular weight in the range of lOO,OOO-500,000. Conversely, amylopectin is a highly branched polymer having a molecular weight in the millions. The branches in amylopectin contain about 20-25 glucose
*On leave from Materials Science Centre, Indian Institute of Technology, Kharagpur 721302, India. **This article reports the results on research only. Mention of a pesticide in this article does not constitute a recommendation for use by the U.S. Department of Agriculture nor does it imply registration under FIFRA as amended. The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.

units. Through plant breeding, three types of cornstarch are available commercially; waxy ( < 1% amylose), pearl ( - 25% amylose), and high amylose (50-70% amylose). These starches and mixtures thereof were evaluated as controlled release matrics. It is well known [a-12] that amylose undergoes retrogradation, which is the formation of aggregates resulting from hydrogen bonding between hydroxyl groups of adjacent starch chains. By mixing waxy, pearl and high amylose starch the amylose content can be adjusted to allow different degrees of retrogradation. By controlling the degree of retrogradation we can control the release of bioactive agents from a non-chemically modified starch matrix.
MATERIALS a. Chemicals AND METHODS

Starches from CPC International, wood Cliffs, NJ (pearl), American

EngleMaize-

34

Products, Hammond, IN (waxy), and National Starch and Chemical Co., Bridgewater, NJ ( Amylon V and VII), and flour from Illinois Cereal Mills, Paris, IL (corn flour) were used for all encapsulations. EPTC (S-ethyl dipropylthiocarbamothioate) (98% ) and butylate [2-ethyl bis (2-methyl-propyl) carbamothioate] (97.8% ) were technical grade (Stauffer Chemical Co., Richmond, CA). b. Steam injection (jet) cooking

ml) at 30C and gently stirred several times during the first three hours to prevent clumping. After 24 h the height of the swollen product in the cylinder was measured. 2. Percent butylate encapsulated Hexane-washed samples (0.100 g ) in 2 N HCl (10 ml) covered by isooctane (10 ml) were heated at 100 C for 5 min to dissolve the starch. After cooling, samples were extracted with isooctane and diluted to 25 ml. Butylate concentration was measured in a Tracer 560 gas-liquid chromatograph (Austin, TX) with a Chromosorb WHP column (6 ft, 0.085 in. I.D.). A standard curve was made with EPTC as an internal standard. The unwashed samples were also analyzed by this procedure to determine total butylate recovered (surface and encapsulated). 3. Butylate release studies Samples (0.05 g) and water (45-81 ml) in 125-ml Erlenmeyer flasks were agitated in a horizontal shaker (Eberbach, Ann Arbor, MI) at 120 shakes per minute. Water volume was 20% in excess of that required to dissolve all the butylate in the product (butylate solubility is 45 mg/l water at 22 C-Herbicide Handbook). Experiments were terminated at various times between 15 min and 240 min by extracting the water phase with isooctane ( 25 ml). Butylate concentrations were determined by GC with EPTC as an internal standard.

Aqueous slurries (5 to 45% wt/vol) of cornstarch or mixtures of cornstarches (90 g) were passed through a laboratory-model continuous steam injection cooker (Penick and Ford, Ltd., Cedar Rapids, IA) at 90 to 155 C and at a flow rate of about 1.3 l/min. The amount of steam entering the cooker was used to regulate the desired dispersion temperature and the back pressure was kept constant. Final pastes were diluted by as much as 100% due to steam condensation. c. Butylate encapsulation

The starch paste from the cooker was collected in a Dewar flask and transferred to a Sigma blade mixer. Butylate (10 g) was added to the still hot starch paste (95C) and allowed to mix at 81 rpm for 30 min. Following mixing, the semi-solid mass was dried either on trays in a hood at 28 C overnight or in a forced-air oven at 130C for 90 min. The mass dried to a film, which was ground in a Waring blender. Samples were screened, and the 20-40 mesh fraction was taken for analysis and evaluation. Samples for the encapsulation analysis were washed three times with hexane to remove butylate from the surface of the ground particles. d. Product analyses 1 . Swellability Encapsulated samples (0.20 g) were placed in a lo-ml graduated cylinder with water (4.0

RESULTS Effect of amylose content

Several butylate-encapsulated products were prepared with either waxy, pearl, or high amylose starch or mixtures of waxy and pearl starch. Corn flour also was used to encapsulate butylate. Table 1 summarizes data of product formulation and analysis. As the amount of amylose in the matrix increases the amount of

35 TABLE 1 on encapsulation amylose ratio of butylate and properties Butylate recoveredh 100% 94% 100% 93% 89% 89% 84% 74% 51% of the matrices Butylate encapsulated 58% 50% 74% 56% 53% 77% 71% 62% 41% Swellability H,Od disperses disperses disperses partially 380% 320% 280% 240% 190% 180% in

Effect of starch composition Sample No. 1 2 3 4 5 6 7 8 9

Amylopectin:

100:0 (waxy) 95:5 9O:lO 85:15 SO:20 75:25 (pearl 1 75:25 (corn flour) 50:50 ( Amylon V) 30:70 ( Amylon VII)

90 g starch in 180 ml H,O; 10 g butylate; jet cooked at 135 C; air dried. bTotal butylate recovered in the dry product. bButylate remaining after extracting surface butylate in ground product. dSample (0.2 g) in H,O (4 ml).

100 g 80

95AP. 90AP

5A :lOA

DISCUSSION

p 2 s t? 2 2 2

60 85A.P 15A 80 A.P. : 20 A Pearl 75 A.P 25 A Corn Flour 75 A.P 25 A Amylon V 50 A P : 50 A Amylon VII 30 A P 70 A

40 20

60

120 Time (m(n)

180

240

Fig. 1. Effect of amylopectin rate of butylate release.

: amylose (AP:A)

ratio on the

butylate recovered (surface and encapsulated) decreases and the product swellability decreases. Rate of butylate release was measured in a strenuous water test using a 20% excess of water over that required to dissolve all the butylate in a 0.05-g sample of product. Figure 1 shows that increasing the amount of amylose in the encapsulated matrix reduced the percent of butylate release with time. Corn flour having the same amylopectin:amylose ratio as pearl starch and also containing about 6% protein releases butylate slightly slower than pearl cornstarch.

The encapsulating ability of starch depends on the amylopectin and amylose contents in the matrix. The higher the amylopectin content in the gelatinized starch, the greater is the encapsulating ability. The highly branched nature of amylopectin reduces the mobility of the starch chains and interferes with their tendency to become aligned for close packing. As a result, amylopectin provides a relatively larger volume for the active agent to be accomodated inside the polymer matrix than does amylose (Table 1). After entrapment and during drying, the amylose chains retrograde to form a tighter matrix; in the process they force some of the entrapped active agent molecules from the matrix. This phenomenon explains why waxy starch has the highest encapsulating ability (butylate recovered), Amylon VII the lowest, with pearl starch intermediate between the two. On the other hand, the rate of release of the entrapped butylate from the starch matrix depends on how tightly the active agent is held by the host matrix (Fig. 1) . Since the essentially linear nature of amylose allows the chains to pack closely (retrogradation) , the active agent in the higher amylose-containing matrix is more securely

36

held than in the higher amylopectin-containing matrix. The greater level of branching in the amylopectin matrix allows entrapped molecules to migrate out easily when water is present. With greater amylose content it becomes difficult for the molecules to escape. This is consistent with the experimental results in that the rate of butylate release from the starch matrix follows the order waxy> pearl > Amylon V > Amylon VII. The same order of release was observed for matrices with amylose contents of 5, 10, 15, and 20% prepared from mixtures of waxy and pearl strach. The tightness of an encapsulating matrix can be estimated by its degree of swelling in the leaching medium. This is consistent with the highest swellability for waxy starch and the lowest swellability for Amylon VII. In fact, when the amylopectin content of the starch or mixture of starches is 90% or greater, little retrogradation occurs, and the product nearly completely disperses in water in a short time (Table 1). Keeping other factors constant, swelling of the starch matrix product is a measure of its amylopectin content. Steam injection (jet) cooking [ 1 ] involves direct contact of steam with starch slurries in order to cause granule rupture and starch dissolution. In our study, the best temperature range to get good dispersion with little molecular breakdown was loo-120 aC for waxy starch, 120-135 C for pearl starch and 150-155 C for Amylon VII. Since the gelatinization of starch takes place at neutral pH, the pH of the encapsulated product is determined mostly by the pH of the active agent. Increasing amylose content also affects the maximum amount of active agent that can be entrapped. The maximum amount of butylate (as a percentage of starch by weight) entrapped by waxy starch was 25%, by pearl starch 20% and by Amylon VII 5%. Higher active agent loadings resulted in more butylate being forced to the matrix surface and less entrapment. Oven and air drying were evaluated for product recovery. Oven drying at 130 aC causes little

or no loss of active agent. Slow drying in ambient air allows for more extensive retrogradation which in turn results in a slower release rate of butylate.

CONCLUSION Jet-cooked cornstarch is an effective encapsulation matrix for active agents, such as butylate. The amylose content is important in determining the efficiency of encapsulation and rate of release of active agent. The higher the amylose content the lower the efficiency of encapsulation and the slower the release of butylate. The ability of the linear amylose molecules to retrograde is the governing factor for using starch in controlled-release applications. REFERENCES
R.E. Wing, S. Maiti and W. M. Doane, Effectiveness release

of jet-cooked pearl cornstarch as a controlled matrix, Starch/Starke, in press.

N.N. Hellman, B. Fairchild and F.R. Senti, The bread staling problem. Molecular organization of starch upon aging of concentrated starch gels at various levels, Cereal Chem., 31 (1954) 495-505. T.F. Protzman moisture

and R.M. Powers, Starch polymers and

their use in paper coating, in: R.R. Myers and J.S. Long (Eds.) , Film Forming Compositions, Marcel Dekker. New York, NY, 1968, Chap. 2. R.G. McIver, D.W.E. Elton, Kinetic ized starch, Axford, K.H. Colwell and G.A.H. of gelatin560-563. study of the retrogradation

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U. Matsukura, A. Matsunaga and K. Kainuma, Structural studies on retrograded normal and waxy starches, J. Jpn. Sot. Starch Sci., 30 (1983) 106-113. R.R. de1 Rosario and C.R. Pontiveros, Retrogradation of some starch mixtures, Starch/Starke, 35 (1983) 86-92. V.H. Kriisi and H. Newkom, Investigations on the re-

trogradation in concentrated wheat starch gels. Part 1. Preparation of concentrated gels, influence of the starch concentration and conditions of preparation on the starch retrogradation, Starch/Stlrke, 36 (1984) 40-45. M.J. Miles, V.-J. Morris and S.G. Ring, Some recent observations on the retrogradation of amylose, Carbohydr. Polym., 4 (1984) 73-77.

37

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