Formulation and Evaluation of Rutin Loaded Nanosponges

 

K.Vijaya Sri^*, Guda Santhoshini,  D. Ravi Sankar, K.Niharika

Malla Reddy College of Pharmacy, (Affiliated to OsmaniaUniversity), Secunderabad, Telangana

 

 

ABSTRACT:

The   main objective of this research was to develop  rutin nanosponges   and evaluate the potential of nanosponges  for the delivery oral  route. Rutin exhibited as antioxidant, anti-inflammatory, anti-allergic and  antiviral activity.  But its,   poor absorption  from the gastrointestinal tract is slow and irregular, probably due to their less solubility  and slow dissolution rate. Rutin-loaded nanosponges were prepared by emulsion solvent diffusion method. Nanosponges were evaluated for various physicochemical parameters like FT-IR, DSC, entrapment efficiency ,SEM, particle size, zeta potential   and in vitro drug release. FT-IR study reveals the there was no physical and chemical  interaction observed between the excipients and drug in combination. Rutin-loaded nanosponges enhanced the drug release when compare to  the  pure rutin.

 

 

 

INTRODUCTION:

The advent of nanotechnology lead to invention of many dosage forms. Effective targeted drug delivery system have been a dream for long time, due to several major drawbacks, a practical approach has been developed for the formation of discrete functionalized particles, which have been termed as ‘Nanosponge’¹.The development of new colloidal carrier called nanosponges has the potential to solve these problems. Nanosponge is a novel emerging technology it can precisely control the rates of controlled drug delivery for topical use. The invention of nanosponges has become a significant step toward overcoming these problems.²

 

Nanosponges are tiny sponges with a size of about a virus, which can be filled with wide variety of drugs. These tiny sponges can circulate around the body until they encounter the specific target site and stick on the surface and begin to release the drug in controlled manner.³ Because the drug can be released at specific target site instead of circulating throughout the body it will give more effective for a particular given dosage. Another important character of these sponges is their aqueous solubility; this allows the use of these systems effectively for drugs with poor solubility.^4,5 Nanosponges are new class of tiny sponges that are about the size of virus, filling them with a drug and attaching special chemical “linkers” that bond preferenticallyto a feature found only on the surface of tumour cells and then injecting them in to the body. These tiny sponges circulate around the body until they encounter the surface of tumour cell where they stick on the surface(or are sucked into the cell) and begin releasing their potent drug in a controllable and predictable fashion. Nanosponges are like a three dimensional network or scaffold, whose backbone is long length polyster. It is mixed in a solution with small molecules called cross-linkers that act like tiny grapping hooks to fasten different parts of the polymer together. The net effect is to form spherically shaped particles filled with cavities where drug molecules can be stored. The polyster is biodegradable, so it breaks down gradually in the body.⁶ The size of nanosponges particles can also possible to control by varying the proportion of cross-linkers to polymer the nanosponge particle can be made larger or smaller. The research has shown that drug delivery system is smaller than 100nm.The nanosponge particles used in the current study were 50nm in size 2. Nanosponges are tiny mesh like structure that may revolutionize the treatment of many diseases and this technology is five times more effective than conventional methods. Nanosponges are made up of microscopic particles with a few manometers wide cavities, in which large variety of substances can be encapsulated. These particles are capable of carrying both lipophilic and hydrophilic substances and of improving the solubility of poorly water soluble molecules. Rutin, a very common quercetin glycoside, was recognized since 1946 to decrease the permeability of capillaries. Rutin acts as antioxidant and exhibited several beneficial effects, such as anti-inflammatory, anti-allergic, antiviral as well as an anticancer activity^7-9 and shown in Figure 1.

 

 

 Figure 1. Structure of Rutin

 

It is suggested that rutin plays a protective role in liver diseases, cataract and cardiovascular diseases  and  the absorption of  rutin from the gastro intestinal tract is slow and irregular, probably due to their very slight solubility in water and slow dissolution rate.  Rutin with cyclodextrins and ,nanoparticles formulations were reported. ^10-14 The   main objective of this  present research was to develop  a simple nanoparticle production technique and evaluate the potential of rutin  loaded nanosponges.

 

MATERIALS AND METHODS:

Rutin purchased from  sigma alderich Other ingredients used were ethyl cellulose (SD Fine Chemicals Ltd, Mumbai, India), Polyvinyl alcohol, Dichloromethane, Triethanolamine (Molychem India), Carbopol 934 NF (Finar Ltd. India), Propylene glycol (Molychem  Ltd, Thane, India) and N-methyl-2-pyrrolidone (Finar Ltd. India). All other ingredients used were of analytical grade.

 

Compatibility Studies:

Fourier transform –infra red:

The pure drug Rutin, polymers like polyvinyl alcohol, ethyl cellulose dichloromethane and physical mixture  were mixed  with IR grade KBr. The powder blends  were scanned over a wave number range of 500 to 4000 cm-1 and studied by FTIR Bruker 160A, (Kyoto, Japan)  model instrument.

 

Preparation of Emulsion solvent diffusion method:

Nanosponges were prepared by using different ratios of ethyl cellulose and polyvinyl alcohol directly prepared by emulsion solvent diffusion method.² In this method, two phases, the continuous phase comprised of a specified amount of polyvinyl alcohol  in 150 mL water and the disperse phase consisted of rutin (100 mg) and definite quantity of ethyl cellulose dissolved in 20 mL dichloromethane. The organic dispersed phase was added slowly in to the aqueous continuous phase at 35°C and reaction mixture was stirred at 1000 rpm for 2 hr on a magnetic stirrer. The nanosponges formed were collected and dried in an oven at 40°C for 24 h and stored in dessicator to ensure the removal of residual solvent.

 

Percentage entrapment efficiency:

A specified quantity (10 mg) of nanosponges was placed with 5 mL of methanolicHCl  in a volumetric flask and was shaken for 1 min vigorously. The volume was made up to 10 mL using methanolicHCl. The solution was filtered, appropriately diluted and the concentration of rutin  was determined spectrophotometrically  at 220 nm. The measurement of percentage drug content of each formulation was carried out in triplicate and the average values are reported.

 

Scanning Electron Microscopy:

The materials were placed onto carbon plates and coated under an argon atmosphere with gold to a thickness of 5mm.

 

Particle size, polydispersity index and zeta potential determination:

Nanosponges properties such as particle size diameter, zeta potential and size distribution were determined by HORIBA scientific nanopartica (Nano particle size analyzer) SZ-100. The polydispersity index (PDI), which is the width of the particle size distribution curve, was determined as a measure of the homogeneity. Polydispersity of rutin loaded nanosponges non uniform size was calculated from the formula,    

Polydispersity = [D0.9 - D0.1] ÷ D_0.5

 

Where D_0.9, D_0.1 and D_0.5 are particle diameters determined at 90th, 50th and 10th percentile of undesired particles respectively. Small values of PDI indicate a homogeneous population, while high values of PDI indicate its high heterogeneity.

In vitro drug release:

The in vitro release study of rutin  from the nanosponge equivalent to 10 mg of drug was determined using USP paddle apparatus. The paddle rotation speed was kept at 50 rpm and a temperature of 37 ± 0.5°C was maintained. A release study was carried out in 900 mL of phosphate buffer pH 6.8 as dissolution medium separately. Five mL of sample was withdrawn at predetermined intervals and replaced by its equivalent volume of fresh dissolution medium to maintain the sink condition. The withdrawal liquids were filtered and assayed at 220 nm for study at pH 6.8.

 

Differential scanning calorimetry:

DSC analysis of rutin and  different ratios of nanosponges  each sample under the analogous conditions of temperature range 0 – 420º C, heating rate 20ºC/min, nitrogen atmosphere (20ml/min) and alumina as reference. DSC measurements were done on a Shimadzu DSC-60 and samples were heated.

 

RESULTS AND DISCUSSION:

Fourier transform –infra red:

No compatibility between drug and excipients and finally confirm that there was no chemical modification.

 

Entrapment Efficiency:

Encapsulation efficiency is the percentage of drug loaded within the polymeric nanoparticle compared to the actual or theoretical mass used for the nanoparticle preparation. It is heavily dependent on the polymer–drug combination as well as the method deployed.The entrapment efficiency was found to be in the range of  56.7%  to 81.6%  for all the nanosponges.

 

Scanning electron microscopy:

SEM indicated uniform and spherical shape, discrete particles without aggregation and appear to be smooth in surface morphology with nano-size range. The nanosponges are spherical, uniform, sponge like structures are shown in the figure 2.

 

Figure 2.SEM Photo graphs of a.NF1 b.NF2 c.NF3

 

Particle size, Zeta potential and Polydispersity index:

All formulations produced particles of size less than 1000 nm, and hence met the requirements to be characterized as nanoparticles. The zeta potential is an important parameter when considering the stability of the nanoparticles in vitro. The more negative or positive values of zeta potential are related to more stable particles, more repulsion between particles reduce the particle aggregation. PDI is a measure of the heterogeneity of particle size for a given emulsion/suspension. A relatively low PDI value (less than 0.1) indicates a good quality narrow particle size distribution range16 while a high PDI value (value towards 1.0) indicates a lower quality broad particle size distribution range. A lower PDI is also desired over a higher PDI, as the particles would be more monodispersed hence a lower likelihood of aggregation and subsequently a decreased chance of sedimentation. This aids the bioavailability aspect of the drug after clinical administration. Particle size  and  zeta potential of optimized formulation (NF2) was determined and found 48.3 nm and -17.2mV indicating low particle size therefore greater surface area and high absorption indicating the formulation is stable and mention the  table 1.

 

 

Table 1. Different formulations of Particle size Diameter (nm) Zeta potential, PDI and Entrapment efficiency (%)of rutin loaded nanosponges

Formulation code	Particle size  Diameter (nm)	Zeta potential (mV)	PDI	Entrapment efficiency (%)
NF1	60.40	-12.8	1.000	56.7
NF2	48.30	-17.2	1.000	81.6
NF3	54.04	-14.6	0.986	75.1

 

 

 

In vitro drug release:

The in vitro release profiles of the formulated rutin nanosponges in pH 6.8 are depicted in fig 3. The drug release from all the formulations was higher in pH 6.8 (simulated pH of normal skin) due to higher solubility of drug in phosphate buffer pH 6.8. Among all the prepared nanosponges,NF2 made with PVA:EC (3:2) exhibited highest drug release.

 

Figure 3 Percentage of drug releases of rutin loaded nanosponges

Differential scanning calorimetry:

The DSC thermogram of rutin exhibited sharp endothermic peaks at 143.62⁰C and at 190.05⁰C indicating the melting point of rutin  and the same was retained in the optimized  nanosponges indicative of no change in the state of the drug.

 

 

Figure 4. Differential scanning calorimetric of optimized nanosponge formulation

 

CONCLUSION:

Nanoparticles have an increased surface area to volume ratio compared to microparticles defining why they have increased dissolution rates and hence enhanced bioavailability. Rutin loaded nanosponges  were prepared by emulsion  solvent diffusion method   and evaluated. The formulation NF2 having less particle size and  high   drug release when compared with the other  two formulations and  NF2 formulation showed optimized activity.

 

ACKNOWLEDGEMENTS:

Authors are grateful to authorities of  Malla Reddy College of  Pharmacy, for providing  the facilities to  conduct  this research work.

 

REFERENCE:

1.     S.P. Vyas, R.K. Khar. Targeted and controlled drug delivery-novel carrier systems: molecular basis of targeted drug delivery. CBS Publishers and Distributors; New Delhi: 2012: 38- 40.

2.     Patricia Vsan Arnum: Nanosponges, a controlled – release Nanoparticle system, shows promise in targeted drug delivery. Pharmatech.com. 35(5): 2011; 56-60.

3.     Santanu K, Sabyasachi M, Ashoke Kumar G, Dilipkumar Pal, Animesh G, and Subham Banerjee  Microsponges: A novel strategy for drug delivery system Journal of  Advance pharmaceutical  technology and research. 2010; 1(3): 283–290

4.     S S, S A, Krishnamoorthy K, Rajappan M. Nanosponges: a novel class of drug delivery system—review. Journal of Pharmacy and Pharmaceutical Sciences. 2012;15(1):103-11

5.     Sharma R, Roderick B and Pathak K: Evaluation and kinetics and mechanism of drug release from Econazole Nitrate Nanosponges loaded carbopol hydrogel. Indian Journal of Pharmaceutical Education and Research. 2011;45(1):25-31

6.     Saija, Scalese M, Lanza M,etal. Flavonoids as  antioxidant agents: Importance of  their interaction  with biomembranes. Free Rad. Biol. Med.,1995; 19, 481-486

7.     Miller A.L.  Antioxidant flavonoids: structure, function and clinical usage. Alt. Med.  Rev., 1996; 1: 103-111.

8.     Alcaraz M. J., Ferrandiz M. L.:  Modification of arachidonic metaboilism by flavonoids. J. Ethnopharmacol., 1987;21: 209-29.

9.     Wilson RH, Mortarotti TG, De EF. Some pharmacological properties of rutin. J Pharmacol Exp Ther 1947;90:120–7.

10.   Miyake K, Arima H, Hirayama F, Yamamoto M, Horikawa T, Sumiyoshi H, et al. Improvement of solubility and oral bioavailability of rutin by complexation with 2-hydroxypropyl-betacyclodextrin. Pharm Dev Technol 2000;5:399–407.

11.   Vijaya Sri,K Kondaiah ,A  VijayaRatna J and  Annapurna A.;  Preparation and characterization of Quercetin and Rutin cyclodextrin inclusion complexes. Drug Development and Industrial  Pharmacy. 2007;33(3): 245-253.

12.   RababKamelDina M.Mostafa Rutin nanostructured lipid cosmeceutical preparation with sun protective potential J.of Photochemistry and Photobiology B: Biology.2015;153:59-66

13.   RababKamel  MonaBasha Preparation and in vitro evaluation of rutin nanostructured liquisolid delivery system. Bulletin of Faculty of Pharmacy, Cairo University 2013; 51 (2):261-272

14.   .Mauludin R, Muller RH, Keck CM. Development of an oral rutin nanocrystal formulation. Int J Pharm 2009;370:202–9

 

 

 

 

 

 

Accepted on 11.03.2018            © A&V Publications All right reserved