USE OF MALTODEXTRIN AS AN EXCIPIENT

Abstract

The present invention relates to the use of maltodextrin as an excipient. In particular, the present invention relates to the use of maltodextrin as an excipient in probiotic formulations.

The present invention also relates to a method to provide a probiotics formulation with improved dispersibility and/or shelf life comprising adding maltodextrin to probiotic formulations.

Claims

1-6. (canceled)

7. The method according to claim 11, wherein the maltodextrin is further combined with a base.

8. The method according to claim 7, wherein the base is selected from Sodium hydroxide (NaOH), Potassium hydroxide (KOH) or Ammonium hydroxide (NH.sub.4OH).

9-10. (canceled)

11. A method to improve shelf life of freeze-dried probiotic bacteria comprising: blending maltodextrin with the freeze-dried probiotic bacteria to form a blend, and storing the blend, wherein said maltodextrin has a dextrose equivalent (DE) between 17 and 23 and a water activity (AW) below 0.1.

12. (canceled)

13. The method according to claim 11, wherein the freeze-dried probiotic bacteria comprises Lactobacillus acidophilus.

14. The method according to claim 11, wherein the freeze-dried probiotic bacteria comprises Bifidobacterium lactis.

15. A method for improving shelf-life of a probiotic formulation comprising freeze-dried probiotic bacteria comprising: adding maltodextrin to the formulation, and storing the formulation, wherein: the formulation is in powder form, and the maltodextrin has a dextrose equivalent (DE) between 17 and 23 and a water activity (AW) below 0.1.

16. The method according to claim 11, wherein the maltodextrin is potato maltodextrin.

17-18. (canceled)

19. The method according to claim 15, wherein the maltodextrin is potato maltodextrin.

20. The method according to claim 15, wherein the freeze-dried probiotic bacteria comprises Lactobacillus acidophilus.

21. The method according to claim 15, wherein the freeze-dried probiotic bacteria comprises Bifidobacterium lactis.

22. The method according to claim 15, wherein the method further comprises adding a base to the formulation.

23. The method according to claim 22, wherein the base is selected from Sodium hydroxide (NaOH), Potassium hydroxide (KOH) or Ammonium hydroxide (NH.sub.4OH).

24. The method according to claim 15, wherein the maltodextrin has a dextrose equivalent (DE) between 17 and 22.

25. The method according to claim 15, wherein the maltodextrin has a dextrose equivalent (DE) between 17 and 21.

26. The method according to claim 15, wherein the maltodextrin has a dextrose equivalent (DE) between 17 and 20.

27. The method according to claim 15, wherein the maltodextrin has a dextrose equivalent (DE) between 18 and 20.

28. The method according to claim 15, wherein the maltodextrin has a dextrose equivalent (DE) between 19 and 20.

29. The method according to claim 15, wherein the maltodextrin has a water activity (AW) below 0.09.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0043] FIG. 1: Dissolution profile of different polysaccharides.

[0044] FIG. 2: Aw impact on the Lactobacillus acidophilus (NCFM) stability.

[0045] FIG. 3: Aw impact on the Bifidobacterium lactis (strain BBL) stability.

[0046] FIG. 4: Aw impact on the Bifidobacterium lactis (Strain BBL) stability.

[0047] FIG. 5: Cell recovery at 30 C after 30 days for various maltodextrin pH for various strains of probiotics.

EXAMPLES

Example 1. Transparency and Dissolution Rate of Polysaccharide Excipients

[0048] Various polysaccharide excipients were obtained for transparency and dissolution rate evaluation.

1.1 Materials

[0049] The products that were tested are shown in Table 1 which include potato maltodextrin at various levels of depolymerization (or dextrose equivalent (DE)), some rice maltodextrin at various levels of depolymerization, microcrystalline cellulose, and potato starch.

TABLE-US-00001 TABLE 1 Evaluated polysaccharide excipients Polysaccharides tested Suppliers Characteristics MD 2 AVEBE Potato maltodextrin DE 2 MD 6 AVEBE Potato maltodextrin DE 6 MD 10 AVEBE Potato maltodextrin DE 10 MD 20 AVEBE Potato maltodextrin DE 20 MCC AVICEL microcrystalline cellulose Instance Primera Foods Rice agglomerated maltodextrin (DE 16?) Benefiber Benefiber Wheat dextrin Prejel PA 5 PH DFE Pharma Fully pregelatinised potato starch Maltrin Or R170 Grain processing Rice maltodextrin (DE=12) corporation Rice trim 35 Primera Foods Rice maltodextrin (DE=35) Solani amylum AVEBE Potato starch

1.2 Methods

[0050] The transparency was evaluated by measuring the optical density of solution in which the excipients was fully dissolved and the dissolution rate was tested by measuring the optical density (OD) as a function of time. The method described below simulates what happens when excipients are dissolved into a glass of water.

[0051] A small beaker wrapped with an aluminum foil was filled with 200 ml of tap water at 25° C. and stirred at 600 rpm with a cross-shaped bar activated by a magnetic stirrer. The turbidimeter ASD19-N from Optek inline control (Optek-converter FC20 with storage time interval 1s) was installed on the side of the container. 3.5 gr of sample was added on the surface of the liquid.

[0052] The transparency of the dissolved excipients was evaluated by measuring the optical density after 600 sec (or 10 minutes) of stirring.

[0053] The dissolution rate was followed by measuring the OD as a function of time. The results were expressed as a percent of OD divided by the maximum OD obtained during the run and also expressed as OD for various mixing times.

1.3 Transparency results of dissolved excipients

[0054] Table 2 shows the optical density of the various excipients after 600 sec of dissolution in water.

TABLE-US-00002 TABLE 2 OD after 600 sec of dissolution for various excipients Polysaccharides OD after 600 tested s MD 2 0.011 MD 6 0.022 MD 10 0.037 MD 20 0.000 MCC 1.300 Instance MR211 0.135 Benefiber 0.006 Maltrin OR R170 0.000 Rice trim 35 0.057 Solani amylum 0.794 Prejel PA 5 PH 0.004

[0055] As shown in Table 2, the four most transparent dissolved excipients are MD 20, Benefiber, Maltrin R170, and Prejel. The dissolution rates of those excipients were then measured.

1.4 Dissolution Rates Results

[0056] FIG. 1 shows the percent of OD measured every second divided by the maximum OD as a function of time for the four excipients with the best transparencies in water.

[0057] In a first step, the percent OD increases very rapidly corresponding to a phase during which the powder is dispersing in the water column, then in a second phase the OD percent drops as the excipient is being dissolved. The percent OD drops very quickly for fast dissolving excipients like MD 20, other like Prejel PA 5PH the powder creates some lumps during the dispersion phase; the lumps then absorb strongly when passing in front of the OD reader which explains the successive peaks of absorption.

[0058] The OD measurement at various stirring time are then reported in Table 3 for all the excipients

TABLE-US-00003 TABLE 3 OD in the water column at various stirring time Polysaccharides Time (s) tested 60 120 160 180 MD 2 0.080 0.063 0.053 0.047 MD 6 0.164 0.175 0.161 0.153 MD 10 0.112 0.136 0.117 0.117 MD 20 0.006 0.001 0.000 0.000 MCC 1.044 1.215 1.240 1.246 Instance 0.128 0.134 0.135 0.135 Benefiber 0.033 0.022 0.018 0.016 Prejel PA 5PH 0.653 0.791 0.399 0.054 Maltrin OR R170 0.060 0.027 0.020 0.017 Rice trim 35 0.590 0.038 0.034 0.034 Solani amylum 0.642 0.728 0.739 0.744

[0059] Table 3 clearly shows that the potato maltodextrin with a dextrose equivalent of 20 (MD20) has the fastest dissolution rate and the highest transparency.

Example 2: Transformation of Potato Maltodextrin with a DE of 20 to Improve Probiotic Shelf-Life Stability

[0060] Potato maltodextrin with a DE 20 is clearly the best polysaccharide excipient for dissolution in water or in the mouth. However, the Activity of water (Aw) and acidity measured as pH of MD 20 may not be at its optimum for shelf-life stability. Some additional transformation may need to be done to improve those parameters and improve probiotic shelf-life when mixed with MD 20.

2.1 Method

2.1.1 Aw Measurement

[0061] An Aqualab 4TE, Decagon was used for the measurements of water activity. The sample (about 1 g) was equilibrated within the headspace of a sealed chamber containing a mirror, an optical sensor, an internal fan and an infrared temperature sensor.

2.1.2 pH Measurement

[0062] A C6010 Consor was used for the measurement of pH. A solution was prepared at 100 g/I using demineralized water at 25° C.

2.1.3 Method to Change the pH of Maltodextrin

FB Equipment Description

[0063] All trials were carried out on a GLATT Procell fluid bed dryer. The apparatus was equipped with the GF3 insert and the nozzle (1.2 mm of diameter) was placed in a top position. The inlet air used was taken from a clean room air with controlled humidity (40% RH) and was dehumidified at 1 g H2O/Kg air with a dehumidifier Munters L180E.

Process Parameters and Formulations Used

[0064] The process parameters and formulations are descripted in table 4.

TABLE-US-00004 TABLE 4 Process parameters and formulations used % Mass Spray Nozzle Air Inlet Product Post Sample solutionr MD20 rate pressure flow temperature temperature drying time number added type of solution loaded (g/min) (bar) (m3/h) (° C.) (° C.) Yield (min) 15186B 25% Demineralized water 2000 13.8 2.5 100 68 45.5  5% 24 15186E 22% 1% NaOH 1N 2000 13.5 2.5 100 68 43  8% 8 15202A 26% 25% NaOH 1N 2000 13.5 2.5 100 68 46 19% 22 15203A 28% 1.86% NaOH 1N 2000 15.4 2.5 100 68 43 22% 24 15204A 25% 3.66% NaOH 1N 2000 14.7 2.5 100 68 46 10% 24 15204B 28% 3.1% NaOH 1N 2000 14.8 2.5 100 68 46.6  6% 23

[0065] The final MD 20 pH's obtained are shown in table 5 below.

TABLE-US-00005 TABLE 5 MD 20 pH obtained Water activity Sample number (Aw) pH 15186B 0.078 4.95 15186E 0.088 5.84 15202A 0.087 10.14 15203A 0.09 6.81 15204A 0.073 7.44 15204B 0.076 7.23

[0066] The data demonstrates that by the use of the described process it is possible to make potato maltodextrin with a pH varying from 4.95-10.14

2.1.4 Method to Change Aw

[0067] Samples with various water activity levels were prepared in the fluidized bed reactor by adjusting the inlet temperature and drying time, as shown in Table 6. pH is held constant at pH 7.2.

TABLE-US-00006 TABLE 6 Water activity MD 20 samples obtained Water Sample activity number (Aw) 1 <0.025 2 0.034 3 0.093 4 0.138 5 0.181 6 0.222

[0068] It is thus demonstrated that it is possible to prepare a potato maltodextrin with an Aw varying from 0.025-0.222 by the use of the described process.

2.1.5 Accelerated Shelf-Life Method

[0069] The experiments were conducted with the following freeze dried probiotics: Lactobacillus acidophilus (NCFM strain), Bifidobacterium lactis (strain BBI), Bifidobacterium lactis (strain BBL). The various probiotic species were blended with MD 20 samples equilibrated at various pH and Aw to make a blend with 30 percent of probiotics. The blends were mixed by rotation (about 60 tr/min) in plastic bottle during 20 min. Then, the sealed foil bags were filled with the different blends. The preparation of the samples was made in a clean room at 40% RH and 25 deg. C.

[0070] The sealed foil bags with the blends were stored in an environmental chamber at 30° C. for 3 months or in an environmental chamber at 30° C., 65% HR; or in a chamber at 38 C for 2 weeks (known as Parker test). CFU and Aw were measured at time 0, 1, 3 months to evaluate the impact of excipient Aw and pH on stability performance.

Cell Count Method

[0071] 1 g of sample was precisely weighed in a bottle; then sterile peptone water was added into the bottle up to 100 g and was mixed for 5 minutes at 400 rpm using a bench top shaker, then the samples was let to rehydrate for 20 minutes at room temperature and mixed afterward for 5 minutes to obtain a homogenous solution. A 10.sup.−2 dilution from the original sample was obtained.

[0072] Subsequent dilutions were carried out at 1:10 steps and were made by adding 1 ml of the solution to 9 ml of peptone water. The solutions were homogenized at each step during 20 seconds using a Vortex system at maximum speed.

[0073] MRS agar with 1% of cysteine was used to plate cells.

[0074] For accuracy of determination, it is well known in the art that there must be between 30 and 300 colonies on each agar plate. It is therefore important to estimate the cell concentration of the sample to be analyzed in order to achieve countable plates without having to do too many dilutions.

[0075] For each determination, 4 plates were counted: two different volumes of cell suspension were plated and each volume was made in duplicate. Then the number of colonies obtained on the plates were added and divided by the total sum of the volumes of dilution used for these plates.

[0076] The plates were then incubated at 37° C. for 72 hours and colonies counted.

Calculation Method to Determinate the Recovery

[0077] Probiotic recovery rate was expressed in two different ways.

% Recovery=(Colony Forming Unit (CFU) after storage/CFU t0)*100   a) Percent recovery

Log loss=Log (CFU t0)−Log (CFU after storage   b) Log loss

2.2 Experimental Results

2.2.1 Stability as a Function of Aw

[0078] FIGS. 2 to 4 shows the stability of the probiotic cultures Lactobacillus acidophilus (NCFM strain), Bifidobacterium lactis (strain BBL), Bifidobacterium lactis (strain BBI) as a function of Aw, measured at pH 7.2.

[0079] It is clearly demonstrated that when the water activity is higher than 0.1 the recovery percent decreases. However, as an example when the water activity is below 0.1 a 100% recovery is obtained for the Lactobacillus acidophilus strain even after 1 month at 30 C, see FIG. 2.

[0080] The figures demonstrate that the recovery percentage decreases as the Aw increases.

2-2-2 Stability as a Function of pH

[0081] FIG. 5 shows the recovery percent as a function of pH 4.95, 6.90 and 7.25 for Bifidobacterium lactis (strain BBI), Bifidobacterium lactis (strain BBL) and Lactobacillus acidophilus (NCFM strain). It can be seen from FIG. 5 that certain strains like BBI and BBL are sensitive to the pH of the excipients, and that BBL is having an optimal pH at 6.9 and BBI at 7.25.