Objectives: To optimize formulation of curcumin solid dispersion (CSD). Methods: Prepare curcumin solid dispersion using spray drying method and apply Modde 7.0 software to design the experiments and optimize the formulations.
Journal of military pharmaco-medicine No7-2016 APPLICATION OF MODDE 7.0 SOFTWARE TO OPTIMIZE FORMULATION OF CURCUMIN SOLID DISPERSION USING SPRAY DRYING METHOD Vu Binh Duong*; Nguyen Hai Dang Trieu*; Pham Van Hien* Trinh Thanh Hung**; Nguyen Trong Diep*; Pham Thi Thanh Huong* SUMMARY Objectives: To optimize formulation of curcumin solid dispersion (CSD) Methods: Prepare curcumin solid dispersion using spray drying method and apply Modde 7.0 software to design the experiments and optimize the formulations Results: The optimal formulation to prepare curcumin solid dispersion system using spray drying method including: ratio of curcumin: PVP K30 (1:8.8, w/w) composed 0.76% of lutrol F127 and 3.85% aerosil With this optimal formulation, the solubility of curcumin was 6942.47g/L and the dissolution reached 100% in period of 30 minutes Conclusion: The optimal formulation for preparing curcumin solid dispersion system using spray drying method was established that is potential to enhance bioavaibility of this active component * Key words: Curcumin solid dispersion; Spray drying method; Optimization; Experimental design INTRODUCTION Curcumin that is a polyphenol extracted from Radix of Curcuma longa L owns valuable bioactivities as anti-inflammation, antioxidant, inhibition of cancer cell, antimicrobial… [2, 3] In according to biopharmaceutical classification, this compound belongs to IV class with less solubility and penetration [6] In fact, curcumin is nearly insoluble in water, less absorption and fast elimination so its bioavaibility is too low O HO OCH3 curcumin MATERIALS AND METHODS Materials and instruments O H3CO we prepared solid dispersion using spray drying method with carrier as PVP K30 It found that solubility and dissolution were enhanced significantly Based on this research, we continuously optimized compositions of formulation using experimental design software In this publication, we informed the result of formulation optimization of CSD using Modde 7.0 software OH To improve solubility and dissolution rate of curcumin, as the previous publication, Curcumin was supported by Merck Coop PVP K30 passed BP 2008 Other chemicals were complied reagents of pharmaceutical or analysis Modde 7.0 software was provided by Umetrics Inc/MKS Instruments (USA) * Military Medical University ** Ministry of Science and Technology Corresponding author: Vu Binh Duong (vbduong2798@gmail.com) Journal of military pharmaco-medicine No7-2016 CSD was prepared on spray dring centrifugal system LPG5, China; magnetic thermal rotation (Stuart UC 152, UK) Properties of finished product was evaluated on Dissolution Test System (Caleva 10 ST8/1000, UK), UV-Vis spectrometer (Biochrom Libra S70PC, UK); differential Scanning Calorimeter (Setaram DSC 131, France), Powder X-ray Diffraction System (Brucker D8 Advance, Germany), Scanning Electron Microscope (Nova nanoSem 450 FEI) Preparation of curcumin solid dispersion Preparation of CSD was conducted by spray drying method [1] Dissolve PVP K30, lutrol F127, aerosol in 80% EtOH, stirred on magnetic thermal rotater to completely solution This solution was then added curcumin, kept stirring to dissolve completely and sprayed at 1100C, flow rate was set at 15 mL/min, suppress pressure was 0.2 MPa Finished solid dispersion was stored in brownish glass bottle with tightly cap Experimental design and optimization of formulation * Experimental design: In the aim of finding the optimal CSD formulation, we designed experiments using central composite face-centered model on Modde 7.0 [5] As our previous publication, the main factors effected curcumin solubility and dissolution including ratio of curcumin/PVP K30, lutrol F127, aerosil In this study, these were used as factors Solubility (Y) and dissolution of curcumin after 10, 20, 30 mins (Y10, Y20, Y30) were used separately as the responses in the mathematical modeling Table 1: Variants in central composite face-centered design Factors Signal Level Low -1 Target High +1 Curcumin:PVP K30 X1 0.1 0.15 0.2 Lutrol F127 (%) X2 0.5 0.75 Aerosil (%) X3 Response Y10, Y20, Y30 % curcumin liberate on testing medium on period of 10, 20, 30 mins Y Curcumin solubility (mg/l) * Optimization of experiment: After having the result of designing experiment, using Modde 7.0 software to investigate the effect of factors (independent variables) to responses by publishment of quadric equation scheming factors - responses correlation as the following: Yi = b0 + b1X1 + b2X2 + b3X3 + b11(X1)2 + b22(X2)2 + b33(X3)2 + b12(X1 X2 ) + b13(X1 X3) + b23 (X2 X3 ) Beside, Moddle software also predicted the optimal conditions to get the highest solubility and dissolution of curcumin (Yi max) Finally, optimal condition was tested by doing experiment following the prediction of software Journal of military pharmaco-medicine No7-2016 Yi = b0 + b1X1 + b2X2 + b3X3 + b11(X1)2 + b22(X2)2 + b33(X3)2 + b12(X1 X2 ) + b13(X1 X3) + b23 (X2 X3 ) Beside, Moddle software also predicted the optimal conditions to get the highest solubility and dissolution of curcumin (Yi max) Finally, optimal condition was tested by doing experiment following the prediction of software * Investigate properties of CSD: - Solubility: Transfer a saturate amount of curcumin or CSD (eq to 50.0 mg curcumin) to 50 mL volume flask, make to fill with water, sonicate for 24 hours at 250C This mixture was stored at room temperature for 48 hours more, was then filterred via 0.45 µm membrain, diluted in water (if any) Optical density was measured on UV-Vis spectrometry at 423 nm, compared to standard solution of curcumin to calculate its solubility in both of material and CSD [4] - Dissolution test: Dissolution studies were carried out using USP dissolution apparatus and a paddle at a speed of 50 rpm with 900 mL of H2O as dissolution medium at 370C Solid dispersion powders equivalenting or 50 mg curcumin were dispersed on the surface of the dissolution medium and the time was recorded The samples (2 mL) were collected at 10, 20 and 30 mins with replacement by an equal volume of medium The withdrawners were then through a 0.45 µm filter All the readings were blanked with same media as it was used in the dissolution study Curcumin that dissolved in test medium was determinated by UV-Vis spectrometry at 423 nm Based on optical density of curcumin standard solution to calculate curcumin that was solubilized in medium - X-ray diffraction: Powder X-ray diffraction patterns were obtained with a diffractometer (Brucker D8 Advance, Germany) The operating conditions were as follows: scanning angle: - 50o; scanning speed: 0.02o/0.5s; duration: 29.5 mininutes at 250C - Different scanning calorimetry: Thermal analysis of curcumin, PVP K30, CSD and physical mixture of curcumin and PVP K30 which contained same composition of CSD were carried out using differential scanning calorimetry method at heating rate of 5oC/min for 60 minutes and heated from 20 - 350oC - Scanning electron microscope (SEM): SEM was investigated of appearance, size, porosity, alignment of particles on SEM Nova Nanosem 450 RESULTS AND DISCUSSIONS Experimental design and optimization All factors were put on Modde 7.0, and then, this software established design of 17 experiments Preparation of CSD using dry spraying method followed this design Each of experiment, solubility and dissolution of curcumin from CSD were determined at 10, 20 and 30 minutes Journal of military pharmaco-medicine No7-2016 Table 2: Results of experiment design and observed response values Exp Normalised levels of factors(matrix) Factors Solubility (mg/l) Dissolution (%) 10 mins 20 mins 30 mins X1 X2 X3 X1 X2 X3 Y Y10 (%) Y20 (%) Y30 (%) -1 -1 -1 0.1 0.5 5860.39 68.18 83.11 92.92 -1 -1 0.2 0.5 5940.33 64.95 70.4 72.45 -1 -1 0.1 5096.65 65.37 82.53 92.21 1 -1 0.2 3663.38 51.25 63.39 70.47 -1 -1 0.1 0.5 5733.33 49.94 64.35 70.18 -1 0.2 0.5 5472.09 47.01 58.58 57.85 1 0.1 0.1 6445.68 62.19 76.45 85.01 1 0.2 0.1 5262.24 57.65 66.42 67.46 -1 0 0.1 0.75 7003.85 80.81 97.25 100.65 10 0 0.2 0.75 6381.44 67.36 76.17 78.91 11 -1 0.15 0.5 6926.77 69.9 85.07 90.49 12 0.15 6746.9 68.65 82.46 85.59 13 0 -1 0.15 0.75 6325.77 71.58 77.69 85.16 14 0 0.15 0.75 6227.27 64.66 73.75 81.64 15 0 0.15 0.75 6468.52 81.56 89.64 91.62 16 0 0.15 0.75 6698.36 81.25 89.89 94.33 17 0 0.15 0.75 6584.15 79.52 88.05 92.04 The analyzer of obtained data was carried out using ANOVA The parameter was evaluated using F - test and a polynomial equations were generated for responses Mathematically, relationship that was generated using multiple line arregression analysis for the study variables were expressed in term of coded factors as the following: Solubility: Y = 6,853.36 - 279.25X1 - 217.41X2 + 180.16 X3 - 238.41(X1)2 - 204.43(X2)2 411.54(X3)2 - 193.82(X1 X2 ) - 4.82(X1X3) + 280.79 (X2 X3) R2 = 0.920 Dissolution for 10 mins: Y10 = 78.88 - 2.99X1 + 0.42X2 - 3.22X3 - 2.82(X1)2 - 4.6(X2)2 5.65(X3)2 - 1.02(X1 X2) + 0.87(X1X3) + 3.14(X2 X3) R2 = 0.964 Dissolution for 20 mins: Y20 = 88.49 - 5.64X1 + 0.7X2 - 3.14X3 - 2.43(X1)2 - 3.38(X2)2 5.38(X3)2 - 0.85(X1 X2) + 0,.7(X1X3) + 2.46(X2 X3) R2 = 0.942 10 Journal of military pharmaco-medicine No7-2016 Dissolution for 30 mins: Y30 = 92.61 - 7.424X1 + 1.34X2 - 4.02X3 - 2.08(X1)2 - 2.9(X2)2 5.36(X3)2 - 0.59(X1 X2) + 1.2(X1X3) + 2.13(X2 X3) R2 = 0.961 Above equations noted that it had a close correlation between PVP, lutreol F127 and aerosil to corresponding solubility and dissolution rate of curcumin from CSD with all correlation coefficients that were not less than 0.9 It means that, these factors effected solubility and dissolution rate of curcumin from CSD Fig (a,b) showed 3D contour surface plot showing effect of factors to responses (a) Figure 1: 3D contour surface plot showing effect of aerosil, lutrol F127, PVP K30 to solubility of curcumin from CSD The results in fig indicated that the more PVP K30 and PVP K30 increased, the more curcumin solubility of CSD increased The effect levels of PVP K30 and lutrol F127 were the same Meanwhile, increasing - 4% of aerosil improved solubility of curcumin but keeping increasing of this excipient to 5% possessed tendency decreasing of active ingredient solubility in test medium This was explained by lipophilic property of aerosil 11 Journal of military pharmaco-medicine No7-2016 (a) (b) (c) (d) (e) (f) Figure 2: The 3D contour surface and prediction plot showing effect of factors (independent variables) to dissolution of curcumin for duration test of 10 mins (a,b), 20 mins (c,d) and 30 mins (e,f) The results in fig showed that, increasing of PVP K30 improved dissolution of curcumin in all withdrawn points for 10, 20 or 30 mins Raise of lutrol F127 possessed decreasing potency of curcumin dissolution in period of 20 and 30 mins testing 12 Journal of military pharmaco-medicine No7-2016 Dissolution of curcumin archived the highest at 4% aerosil and decreased while raising of this expicient Modde 7.0 also predicted the optimal formulation to prepare CSD including ratio of curcumin: PVP K30 as 1:8.8 (w/w) compose 0.76% lutrol F127 and 3.85% aerosil The theory solubility and dissolution of curcumin were Y = 6,850.25 mg/L, Y10 = 79.77%; Y20 = 92.27%; Y30 = 98.64%, respectively Running experiment followed the optimal formulation to verify prediction of software Table 3: The results of experiments verify prediction of software Y Predicted Y10 (%) Y20 (%) Y30 (%) Actual Predicted Actual Predicted Actual Predicted Actual 6850.25 6999.57 79.77 79.43 92.27 93.26 98.64 100.73 6850.25 6879.66 79.77 80.84 92.27 94.49 98.64 99.55 6850.25 6845.40 79.77 79.54 92.27 92.76 98.64 99.32 6850.25 6999.57 79.77 80.63 92.27 93.74 98.64 100.63 6850.25 7025.27 79.77 80.08 92.27 93.74 98.64 101.60 6850.25 6905.35 79.77 80.19 92.27 93.84 98.64 101.28 X 6850.25 6942.47 79.77 80.12 92.27 93.64 98.64 100.52 p > 0.05 p > 0.05 The results showed that the solubility and dissolution of curcumin from CSD that was prepared following prediction of Modde 7.0, in period of 10, 20 and 30 mins was undifferentiated significantly in comparison to result of verified actual experiment The predicted - actual values were 6,942.47 mg/L and 6,850.25 mg/L in case of solubility These values were 80.12% vs 79.77%, 93.64% vs 92.27% and 100.52% vs 98.64% in terms of dissolution in period of 10, 20 and 30 mins These findings indicated that optimal formulation preparing CSD via spray drying method included the ratio of curcumin: PVP K30 as 1:8.8; 0.76% lutrol F127; 3.85% aerosil p > 0.05 p > 0.05 Property investigation of CSD * Solubility: The solubility testing of curcumin from CSD and original material showed that the value in case of CSD folded 16.500 times in compare to material (6.942.47 mg/L vs 0.42 mg/L) * X-ray diffraction: X-ray diffraction patterns for orginal curcumin, PVP K30 and CSD were detected in fig Curcumin gave numerous sharp narrow and intense peaks indicating its high crystalline PVP gave no sharp narrow peaks and implicated it exist in amorphous In physical mixture, the carrier appeared as an elevated baseline and interacted to 13 Journal of military pharmaco-medicine No7-2016 curcumin producing characteristic diffraction peaks CSD did not show the characteristic peaks indicating reduction in crystalline and phase transition from crystalline to amorphous via formation of hydrogen bonds between curcumin - PVP K30 Fa cu lty o f C hemistry, H US , V NU, D AD VA NC E -Bruker - Cu rcum in nguye n lieu 30 00 Fa cu lty o f C he m istry , H US , V NU , D A D VA NC E -Br uk er - P VP K ng uye n lieu 29 00 10 0 28 00 27 00 26 00 90 25 00 70 d=5.211 24 00 (a) 80 23 00 22 00 (b) 21 00 20 00 19 00 18 00 50 17 00 16 00 15 00 14 00 d=10.230 Lin (Cps) Lin (Cps) 60 13 00 12 00 40 11 00 d=1.898 d=2.116 d=2.005 d=2.046 d=2.399 d=2.205 d=2.274 d=2.494 d=2.457 d=2.855 d=2.600 d=3.186 d=3.278 d=3.356 d=3.850 d=4.239 d=3.762 d=4.960 d=3.087 d=2.693 20 d=4.771 d=4.616 30 d=4.432 40 d=6.169 50 d=6.431 60 d=7.359 d=11.432 70 10 d=5.871 d=5.664 d=5.451 80 d=3.500 90 20 d=3.634 10 00 30 10 0 10 20 30 40 10 20 30 2-T heta - Sc ale 40 2- Theta - Sc ale F ile : T r ie uH V Q Y P V PK 0n g u yen li eu w - T yp e : T h/ T h lo cked - S t art: 8 ° - E nd : 8 ° - S t e p: 0 30 ° - S te p t i m e : s - T e mp : ° C (R o om ) - T im e S t ar t ed : 17 s - 2- T h e t a: 8 ° - T h eta : 00 ° - C hi : 00 ° - Ph F ile : Tr ie uHV Q Y C ur- ng uye n lie u.ra w - Type : 2Th /Th loc ke d - Sta rt: 0.8 85 ° - En d : 9.8 86 ° - S te p : 03 ° - S te p tim e: 0.3 s - Te m p : °C ( Roo m ) - Tim e Sta rte d: 1 s - -Th eta : 0.8 85 ° - The ta: 0 ° - Ch i: 0.0 ° - P h i: 00 -0 09 -0 81 ( Q) - Cu rcu m in - C2 1H2 O6 - Y : 56 % - d x b y: - W L: 06 - Fa cu lty o f C he m istry , H U S , V N U , D A D V A N C E -B r uk er - HH V L C ur -PV P K Fa cu lty o f C he mi stry , H U S , V N U , D AD VA NC E -Bruk er - H P T R C u r-P VP K3 10 00 10 00 90 90 (c) 80 70 60 Lin (Cps) 60 50 d=4.229 20 30 d=3.375 d=3.659 d=9.898 d=5.214 30 50 40 d=3.780 d=6.472 40 d=3.503 Lin (Cps) (d) 80 70 20 10 10 0 10 20 30 40 10 20 30 40 2-Theta - S c ale 2-T heta - S c ale File : T r ie uH V Q Y H PT R C ur -P VP K 30 w - T yp e : Th /Th lo ck ed - S tar t: 88 ° - E n d: 49 88 ° - S t e p: 0 ° - S t ep ti m e : s - T em p : 25 °C (R o om ) - Ti m e S t a rt ed : 11 s - 2- T he t a: 88 ° - Th et a : 00 ° - C hi : 00 ° - P hi F ile : T r ie uH V Q Y H HVL C u r-P V PK 0.ra w - Ty p e: 2T h /Th l oc ke d - Sta rt: 0.8 85 ° - En d : 8 ° - S tep : 03 ° - Ste p tim e: s - T e m p : °C ( R oo m ) - Tim e S tarte d: s - -Th eta : 0.8 85 ° - The ta: 50 ° - Ch i: 0.0 ° - P h i Figure 3: X-ray diffraction patterns of PVP K30 (a), orginal curcumin (b), physical mixture of curcumin: PVP K30 (c) and CSD (4) * Differential scanning calorimetry: Fi gure: Experiment: PVP K30 nguyen l ieu Crucible:Al 100 µl 22/05/2016 Procedure: RT -300 C (Zone 2) DSC131 Fi gure: Atmosphere:Ar Mass (mg): 20.76 Experiment: Cur nguyen li eu Crucible:Al 100 µl Atmosphere:Ar 22/05/2016 Procedure: RT -300 C (Zone 2) DSC131 Mass (mg): 15.23 HeatFlow/mW HeatFlow/mW Exo Exo -10 -5 -20 Peak :190.8678 °C Onset Poi nt :169.7208 °C Enthal py /J/g : 11.7825 (Endothermi c effect) -30 - 10 -40 - 15 -50 (a) - 20 Peak :91.2850 °C Onset Point :42.3669 °C Enthalpy /J/g : 246.2637 (Endothermic effect) -60 (b) Peak :195.6205 °C Onset Point :191.2751 °C Enthalpy /J/g : 128.5615 (Endothermi c effect) -70 - 25 -80 25 50 Fi gure: 75 100 125 150 175 Experiment: HHVL Cur - PVP K30 200 225 Crucible:Al 100 µl 22/05/2016 Procedure: RT -300 C (Zone 2) DSC131 50 Furnace temperature /°C Mass (mg): 20.13 75 Figure: Atmosphere:Ar 100 125 150 175 200 Experiment: HPT R Cur - PVP K30 225 Crucible:Al 100 µl 22/05/2016 Procedure: RT-300 C (Zone 2) DSC131 Furnace temperature /°C Atmosphere:Ar Mass (mg): 13.24 HeatFlow/mW H eatFlow/mW Exo Exo 0 -2 -5 Peak :184.4112 °C Peak :191.6883 °C Onset Point :177.7930 °C Enthal py /J/g : 8.9469 (Endothermi c effect) (3.8781 + 5.0688) -10 -4 -6 -15 -20 -8 Peak :87.0017 °C Onset Point :43.7985 °C Enthal py /J/g : 223.7256 (Endothermi c effect) (c) -25 25 50 75 100 125 150 175 200 225 Furnace temperat ure /°C (d) Peak :82.1471 °C Onset Poi nt :46.0151 °C Enthalpy /J/g : 136.4217 (Endothermi c effect) -10 50 75 100 125 150 175 200 225 Furnace temperature /°C Figure 4: Differential scanning calorimetry of PVP K30 (a), curcumin (b), physical mixture of curcumin: PVP K30 (c) and CSD (d) 14 Journal of military pharmaco-medicine No7-2016 The possible interaction between the curcumin and the carrier was studied by DSC (Fig 4) Curcumin powder showed a melting endotherm at 195.6oC corresponding to it melting point at enthalpy of 138.6 j/g, whereas the scan of PVP-K30 showed a broad endotherm ranging from 80 to 120°C due to the presence of residual moisture In physical mixture, there was a melting endotherm at 191.7oC corresponding to it melting point at enthalpy of 5.1 j/g, less than orginal material of curcumin This indicated that curcumin still existed in crystalline at physical mixture Loss of melting endotherm and enthalpy caused almost curcumin that dissolved in PVP K30 The pattern of CSD showed a melting endotherm between 100 and 250°C, due to loss of water from PVP-K30 and absenced of curcumin peak The absence (a) of curcumin indicated that curcumin was amorphous or was presented as a solid solution inside the PVP-K30 matrix According to these results, the amorphous property of curcumin in the formulation with PVP-K30 was mainly responsible for the dissolution enhancement * Scanning electron microscope: SEM of CSD were measured on SEM Nova Nanosem 450 The particles of CSD that were prepared using spray drying method, were characteristics of regular spherical, smooth surface, centered hollow porosity The diameter of particle was about 10 - 30 µm Original curcumin material was still existed in crystalline clearly (Fig 5) This finding expressed curcumin in CSD transferred to amorphor form instead crystaline as original material (b) (c) ( Figure 5: SEM of CSD (a,b) and curcumin (c) 1:8.8; 0.76% lutrol F127; 3.85% aerosil This CONCLUSION It was applied Modde 7.0 software optimal formulation improved solubility of formulation curcumin and its dissolution in period of preparing CSD using spray drying method 10, 20 and 30 minutes testing archiving The results found the optimal formulation 6,942.47 mg/L, more than 80%, 90% and including the ratio of curcumin: PVP K30 as got 100%, respectively design and optimize REFERENCES 15 Journal of military pharmaco-medicine No7-2016 Duong Quoc Toan, Vu Binh Duong Study on preparation of altretamin using spray drying method Vietnam Pharmaceutical Journal 2015, No 475, pp.19-24 Kumavat Suresh et al Enhancement of solubility and dissolution rate of Curcumin by solid dispersion technique International Research Journal of Pharmacy 2013, (5), Asish K Dutta, Elizabeth Ikiki Novel drug delivery systems to improve bioavailability of curcumin Bioequivalence & Bioavailability 2013, 6, pp.1-9 pp.226-232 Hatcher H et al Curcumin: From ancient medicine to current clinical trials Cellular and Molecular Life Sciences 2008, 65 (11), pp.1631-1652 design for poorly water-soluble drugs based Sartorius Stedim Biotech Company User guide Bio Pat Modde 7.0 2013 Yohei Kawabata et al Formulation on biopharmaceutics classification system: Basic approaches and practical applications International Journal of Pharmaceutics 2011, 420, pp.1-10 16 ... 20, 30 mins Y Curcumin solubility (mg/l) * Optimization of experiment: After having the result of designing experiment, using Modde 7.0 software to investigate the effect of factors (independent... optimization All factors were put on Modde 7.0, and then, this software established design of 17 experiments Preparation of CSD using dry spraying method followed this design Each of experiment, solubility... Preparation of curcumin solid dispersion Preparation of CSD was conducted by spray drying method [1] Dissolve PVP K30, lutrol F127, aerosol in 80% EtOH, stirred on magnetic thermal rotater to completely