X-Ray diffractometry: In order to investigate the physical nature of the encapsulated drug

,
the powder X-ray diffraction technique was used. Crystalline state is another factor influencing
the dissolution and stability behavior of a compound. The crystalline state of the samples was
evaluated to prove the effect of stirring. Upon X-ray examinations, it was observed that the
specific peaks for simvastatin at specified 2θ value in X-RD graph at 10, 15, 17, 20 were not
observed for SNS. This suggested that the crystallinity of simvastatin was not preserved in the
SNS formulation, indicating that the crystalline state of simvastatin was altered following
stirring. The absence of major peaks of simvastatin in case of SNS confirmed formation of
amorphous product which might leads to enhanced solubility of the drug in case of SNS.
Nanoparticles reduced its crystallinity. This is evident from the disappearance of most peaks in
the nanoparticles compared to the drug. X-Ray diffractometry graphs are shown in Fig. 14.

. 10.2468 Int J Pharm Sci Nanotech Vol 7. In vitro dissolution profile of plain drug and SNS batch-7 in pH 1. 2 acidic buffer. Fig. 11. In vitro dissolution profile of plain drug and SNS batches in pH 7.4 phosphate buffer. Issue 2• April-June 2014 Fig.

24 profile of plain d SNS batch-7 469 .Shid et al: Formulation a nd Evaluation o of Nanosuspen nsion Delivery S System for Sim mvastatin vitro Fig. 12. physic al mixture and powder in dis stilled water. In vi dissolution drug.

654 est and stand 2. T he elevated riglyceride le ncreased HDL he above va nanosuspensio profile than th Sci Nanotech K-30. d levels in test mvastatin tre on powder de The simvasta total choles evel more si Lc level than alue it was on powder sh he plaindrug.001). phys Triton X-100. Sodium L sical mixture (3). decreas up to 191. mvastatin pow gnificant decr 8.2470 Fig. while t eceived stan nanosized sim had shown sig evel up to 78 tandard grou 2. erum.28 ± 1. DS SNS (Simv Fig.088 up (plain dru n rats treated ally. the total li the increase w ndard drug.56 ± d group (plain (p < 0. in test grou standard gro in standard um VLDL-c l nd in standar 4 mg/dL. X Batch-7 SN In vivo st successfu the serum with stan to differe SC spectra of (A) astatin Nanosus X-Ray Diffractom NS Powder.191 ± 1. 14.586 mg/dL a hange in lipid group of sim nanosuspensio micactivity. seru est group an 43. The H dard were 35 and 46. LD ndard drug (S ent degrees.307 mg/dL in ntraperitonea ignificantly u with standard 6. Tr altered this e hat were trea (Simvastatin. 13.74±3. study: Injectio ully induced hm TC.Poy pray dried Simva n drug (B) g/kg) has ncreasing reatment elevation ated with T s re nh le st 2 in si w 6 L co le S te 4 te 2 ch gne th tr in th np Int J Pharm yvinylPyroidine astatin (2).049 mg/dL ( LDL-c level i ompared to evels found i Similarly. I . Sp ain Simvastatin X-100 (400 mg a in rats by in DL levels. I ) Simvastatin ph spension) (D) Sim metry of (A) pla on of Triton X yperlipidemia DL-c and VLD Simvastatin) In animals th hysical mixture ( mvastatin (1). TG.

15.08 ± 1.9 respectively. T comparable w The simvasta anti-hyperlip pension redu c.205 mg/dL was found to found in cont mg/dL. The LD ± 1. 2014 ) (C) in that the g/kg TC with 7 ± /kg) evel ared 44 ± rum as DL-c /dL.023 239 mg/dL rt groups was ated rats. Poloxamer-188) as increased d in animals t istration of ent to 20 mg 001) in serum n compared w wed TC 101.50 ± 1 oup (plain dr group 85. ipid level w was prevented Oral admini wder equivale ease (p < 0.86 level 32. L in be trol.383 mg/ 1.43 ± rd group it w HDL-c levels 5.Vol 7. VLDL-c ap < 0.103 mg/dL n drug) having Same dose d up 67.00 mg/dL when g) that show d only with tr se in seru ± 2.001) a tatin drug. .0 riton (400 mg/ um TG le L when compa g value 225. T emonstrated atin nanosusp sterol. 1 ± The with atin ped. LDL-c gnificantly (p plain simvas found that ows greater d It is shown in 2• April-June 2 .4 decreased ser 1.uced and and rom atin ipid .008 mg/dL rug). Issue 2 Luaryl Sulphate. Fr the simvasta decrease in li n Fig. 51.

Plain drug (Simvastatin). *** = Extremely Significant (P < 0.05) .47 ± 2.78 ± 1.20 ± 1.38 ± 8.205*### 43. LDL-Low Density Lipoprotein.828 34.74 ± 3.Shid et al: Formulation and Evaluation of Nanosuspension Delivery System for Simvastatin 2471 TABLE 6 Biochemical analysis of blood sample after induction of hyperlipidemia.32 ± 2.559 TC-Total Cholesterol. # = Non Significant (P > 0.070*** HDL 34.47 ± 1.01).155*** 300. HDL-High Density Lipoprotein.479*** 32.773 282.00 ± 1.84 ± 2.01 ± 9.001).05). # = Control vs.24 ± 1.76 ± 2.001) TABLE 7 Biochemical analysis of blood sample after drug and SNS powder dosing (after 24 hr)/Pharmacodynamic study after 24 hr (after dosing).307***##@@@ TG 134. @@ = Highly Significant (P < 0.66 ± 1.100 55. HDL-High Density Lipoprotein.43 ± 3.Very Low Density Lipoprotein.001).586***### 46.91 ± 2.27 ± 8.15 ± 1.234*** 113.801*** 78. @ = Non Significant (P > 0.39 ± 1. 05).492*** 67. SNS-Simvastatin Nanosuspension.088 38.01).28 ± 1.71 ± 1.822*** TG 130.001). 918 62. VLDL-Very Low Density Lipoprotein. SNS-Simvastatin Nanosuspension.18 ± 2. 315 133.10 ± 3.44 ± 6.17 ± 2. Plain drug (Simvastatin).008### 85.25 ± 1.462*** 64.020 112. * = Non Significant (P > 0.08 ± 1.191 ± 1.26 ± 1.57 ± 3.50 ± 1.438*** 133.239**## Results are expressed as mean ± SEM.86 ± 1.968*** 62.103***### 225.** = Highly Significant (P < 0.84 ± 2.085 35.147*** LDL 82.61 ± 1. ***=Extremely Significant (P < 0. 415 34.59 ± 1.088### 101. Bioanalytical Test Normal Group Mean ± SEM Control Group Mean ± SEM SNS Powder Group Mean ± SEM Plain drug Group Mean ± SEM TC 75. VLDL.07 ± 2.99 ± 1.43 ± 1. TG-Triglyceride. 2 113. TG-Triglyceride.049***###@@@ LDL 65.023 51.48 ± 6. Comparison between the groups was made by one way analysis of variance (ANOVA) followed by Tukey's test TC-Total Cholesterol.654***###@@@ HDL 34.370*** 289. LDL-Low Density Lipoprotein.56 ± 2.344 134.01). * = Normal vs.383***##@@@ VLDL 25. @ = SNS Powder vs Plain drug.744*** 133.279*** 191.115*** 114. @@@ = Extremely Significant (P < 0. ### = Extremely Significant (P < 0. * = Normal vs.479*** VLDL 28. Test Normal Group Mean ± SEM Control Group Mean ± SEM SNS Powder Group Mean ± SEM Plain drug Group Mean ± SEM TC 77. ## = Highly Significant (P < 0. 095 301.83 ± 3.

(A) Pharmacodynamic parameter study after induction of hyperipidemia.(A) (B) Fig. (B) Pharmacodynamic parameter study after dosing of plain simvastatin drug and batch-7 SNS powder. LDL-Low density lipoprotein. . HDL-High density lipoprotein. TC-Total cholesterol. 15. TG-Triglyceride. VLDL-Very low density lipoprotein.

3-D Response surface plot of total drug content was shown in Fig.49% 2 month 91.2472 Int J Pharm Sci Nanotech Vol 7.53% 95. Each batch contains 20 mg of simvastatin.62 × A+0.74% After storage 1 month 92.78% 95. Final Equation in Terms of Coded Factors: PI = +0. two level full factorial designs was adopted for optimization employing the amount of SLS.5000E – 003 × Polo .3 months samples were withdrawn and rested for entrapment efficiency and total drug content.079 × B-0.30000E – 003 × PVPK . a statistical model incorporating interactive and polynomial terms was used to evaluate the responses.026875 × SLS – 6.18–1. 054 × A-0. Total drug content (%) and polydispersity index as dependent variables. 16(B).80875 × SLS+0.05.041 × C Final Equation in Terms of Actual Factors: PI = +98250 – 0. TDC = +93.2.30-5. Y1 is indicating total drug content (%) whereas Y2 is polydispersity index. Concerning polydispersity index. It indicates that this formulation was able to retain its stability up to 3 months. Experimental trials were performed at all 8 possible combinations.38750 – 0. X1= amount of SLS.30% 93.188 The coefficients b1.188.46 – 0. b2.023200 × PVPK – 30 + 0. a 23 factorial design was employed. Issue 2• April-June 2014 Stability study: The stability studies were carried out on optimized formulations. the results of multiple linear regression analysis showed that the coefficients b1 bear a negative sign and coefficients b2 and b3 bear positive sign (R2 = 0. Time period Entrapment efficiency Total drug content Initial 92.29 × B+0. TABLE 8 Stability data of optimized formulation of nanosuspension. the results showed that the coefficients b1. X2 = amount of PVPK-30 and X3 = amount of Poloxamer. and b3 were found to be significantat P < 0.59 × C Final Equation in Terms of Actual Factors: TDC = +95.87% 94. amount of PVPK-30 and Poloxamer-188 as independent variables. 078667 × Polo-188 It can be concluded from the equation (2) that when the concentration of X2 and X3 was increase with decrease in the concentration of X1 then desired total drug content could be obtained and its controlling the stabilization to the nanoparticles for coalescence and high total drug content was observed intheformulation Batch-7. Total Drug Content (TDC).9395).9061) 3-D Response surface plot of polydispersity index was shown in Fig. After 1. The optimized formulation did not show any significant difference in both parameters. b2 and b3 bear a negative sign (R2 = 0. TABLE 9 Design matrix of independent and dependent variable as per 23 factorial design Run X1 (SLS) . 16(A). The samples were stored at 40 ± 2oC and 75% ± 5% RH for 3 months to access their stability.65% Experimental data analysis: A three factor. In order to investigate the factors systematically.91% 3 month 91.

74% 0. Analysis of variance table partial sum of squares-Type III Source Sum of squares df Mean squares F value P-value Probe > F Model 24.01% 0.87% 1117.87% 0.93 1 20.496 67.33% 1916.39 Cor Total 25. Dev.2608 C-Polo-188 2.420 89. TABLE 10 ANOVA for Response Surface Linear Model: Response-1 (Total Drug Content).78% 258. μm-micron meter.62% 903.78 1 2.5% 548.67 1.310 88. 2 B8 8 50 30 91.77% 0.431 70.39 3 8.5-0.4 0.55% 0.X2 (PVPK-30) X3 (Polo-188) Y1 (TDC%) *Partical size (μm) B1 4 25 15 94.6 B3 4 50 15 94.3 0. E *MPD(nm) .3 B4 8 50 15 91. Polo-188-poloxamer-188.9 0. PVPK30-polyvinyl pyrolidine k-30.2 0. MPD-mean partical diameter.2-0.2-0. V% 0.67 1 0.71 0.83% 653.67 Pred RSquared 0.5 B2 8 25 15 90.8 0.17 Adj R-Squared 0.414 74.3-0.5 B5 4 25 30 94.0019 B-PVPK-30 0.69 0.0067 significant A-SLS 20.2-0.45% 0.8% 604.9395 Mean 93.381 92.578 87. EE -entrapment efficiency.3 *Not considered for factorial design.63 R-Squared 0.11% 0.3 B7 4 50 30 95.93 53. 0.87% 624. SLS-sodium lauryl sulphate.7 0.7578 PRESS 6.13 20.6 B6 8 25 30 92.57 4 0.09 0.96 7 Std.689 58.2-0.3-0.9% 0.270 Y2(PI) *%E. TDC-total drug content.0563 Residual 1.7 0.78 7.8941 C.27 0. PIpolydispersity index. nm-nanometer.29 Adeq Precisior 11.8 0.

16. Emulsification Fig.6243 PRESS 0. In this process.0160 Significant A-SLS 0. a poorly water-soluble drug. Means the bioavailability of simvastatin nanosuspension is greater than the plain simvastatin drug due to the decrease in particle size. for the improvement of solubility and dissolution velocity.8356 C.950E-003 4 2. 50 mg PVPK-30.46 Adj R-Squared 0.389 Conclusions Emulsification Solvent Diffusion method was employed to producing nanosuspension of simvastatin. and 30 mg Poloxamer-188 using emulsification solvent diffusion technique at laboratory scale.0692 Residual 8.33 0.029 12. The dissolution of nanosized simvastatin is significantly enhanced compare with the pure simvastatin suspension.014 1 0.17 0. 0. From the In-vivo study value it was found that the simvastatin nanosuspension powder shows greater decrease in lipid profile than the plaindrug.9061 Mean 0. .050 1 0. V% 10.086 3 0. Dev.0092 C-Polo-188 0.036 AdeqPrecisior 10.08 0.86 0.050 22.0325 B-PVPK-30 0. The best nanosuspension of simvastatin can be obtained by 4 mg SLS. by adjusting the operation parameters. Analysis of variance table partial sum of squares-Type III Source Sum of squares df Mean squares F value P-valueProbe > F Model 0.size ranges.023 10.26 Pred RSquared 0.047 R-Squared 0.023 1 0.095 7 Std.238E-003 Cor Total 0. the particle size of simvastatin can be obtained in the nano.Shid et al: Formulation and Evaluation of Nanosuspension Delivery System for Simvastatin 2473 TABLE 11 ANOVA for Response Surface Linear Model: Response-2 (Polydispersity Index).014 6. (A) 3-DResponse surface plot of total drug content (B) 3-D Response surface plot of polydispersity index (PI). such as the stabilizer concentration.

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