Optimization, preparation and characterization of rutin-quercetin dual drug loaded keratin nanoparticles for biological applications

Document Type: Research Paper


1 Sir CV Raman- KS Krishnan International Research Centre, Kalasalingam University, Krishnankoil, India

2 Department of Chemical Engineering, Jadavpur University, Kolkata, India


Objective(s): Response surface methodology (RSM) by central composite design (CCD) was applied to statistically optimize the preparation of Rutin-Quercetin (Ru-Qr) dual drug loaded human hair keratin nanoparticles as well as evaluate the characteristics.
Materials and Methods: The effects of three independent parameters, namely, temperature (X1:10-40 C), surfactant (X2: SDS (1), SLS (2), Tween-20 (3)), and organic solvents (X3: acetone (1),  methanol (2), chloroform (3)) were investigated to optimize the preparation of dual drug loaded keratin nanoparticles, and to understand the effects of dependent parameters namely, drug releasing capacity, average particle size, total antioxidant power, zeta potential, and polydispersity index of Ru-Qr nanoparticles. Optimization was executed by CCD and RSM using statistical software (Design Expert, version, Stat-Ease, Inc., Minneapolis, MN, USA). The optimal Ru-Qr dual drug loaded keratin nanoparticles were obtained at temperature (X1): 40ÚC, SDS (X2), and acetone (X3).
Results:  Under this conditions to achieve highest drug releasing capacity of 98.3%, average size of nanoparticles are 125 nm, total antioxidant power 98.68%, zeta potential 28.09 mV, and polydispersity index of 0.54. Although majority of the experimental values were relatively well matched with the predicted values.
Conclusion: This optimization study could be useful in pharmaceutical industry, especially for the preparation of new nano-therapeutic formulations encapsulated with drug molecules. This nanotechnology based drug delivery system is to overcome multi drug resistance and site specific action without affecting other organs and tissues. The methodology adopted in this work shall be useful in improvement of quality of human health.


1. Heller A. Integrated medical feedback systems for drug delivery. AlChE J. 2005; 51(4): 1054-1066.

2. Roszek B, De Jong WH, Geertsma RE. Nanotechnology in medical applications: state-of-the-art in materials and devices. RIVM report. 265001001/2005.

3. Singh R, Lilard JW. Nanoparticle-based targeted drug delivery. Exp Mol Pathol. 2009; 86(3): 215-223.

4. Selvaraj. K, Chowdhury. R, Bhattacharjee. C. A green chemistry approach for the synthesis and characterization of gold nanoparticles stabilized with methanol extract of Azolla microphylla and their enhanced antioxidant activity. Front Mat Sci. 2014; 8(2): 123–135.

5. Liechty WB, Kryscio DR, Brandon V. Slaughter, Peppas NA. Polymers for drug delivery systems. Annu Rev Chem Biomol Eng. 2010; 1: 149-173.

6. Lohcharoenkal W, Wang L, Chen YC, Rojanasakul Y. Protein nanoparticles as drug delivery carriers for cancer therapy. Biomed Res Int. 2014; 2014: 1-12.

7. Elzoghby AO, El-fotoh WS, Elgindy NA. Casein-based formulations as promising controlled release drug delivery systems. J control release. 2011; 153:206-216.

8. Nie S. Understanding and overcoming major barriers in cancer Nanomedicine. Nanomedicine (Lond). 2010; 5(4): 523-528.

9. Maham A, Tang Z, Wu H, Wang J, Lin Y. Protein-based Nanomedicine platforms for drug delivery. Small 2009; 5(15): 1706-1721.

10. Kakkar P, Madhan B, Shanmugam S. Extraction and characterization of keratin from bovine hoof: A potential material for biomedical applications. Springer plus. 2014; 3: 596.

11.  Feughelmann M. Keratin. Encyclopedia of polymer science and engineering. New York, Wiley.1985.

12. Marshall RC, Orwin DF, Gillespie JM. Structure and biochemistry of mammalian hard keratin. Electron Microsc Rev. 1991; 4(1): 47-83.

13. Parani M, Lokhande G, Singh A, Gaharwar AK. Engineered Nanomaterials for infection control and healing acute and chronic wounds. ACS Appl Mater Interfaces. 2016; 8(16): 10049-10069.

14. Chaudhury K, Kumar V, Kandasamy J, Roychoudhury S. Regenerative nanomedicine: current perspectives and future directions. Int J Nanomed. 2014; 9: 4153-4167.

15. Scarano W, de souza P, Stenzel MH. Dual-drug delivery of curcumin and platinum drug in polymeric micelles enhances the synergistic effects: a double act for the treatment of multidrug-resistant cancer. Biomater Sci. 2015; 3(1):163-174.

16. Zhao XY, Zhu YJ, Chen F, Wu J. Calcium phosphate nanocarriers dual-loaded with bovine serum albumin and ibuprofen: facile synthesis, sequential drug loading and sustained drug release. Chem Asian J. 2012; 7(7):1610-1615.

17. Nakamura A, Arimoto M, Takeuchi K, Fujii T. A rapid extraction procedure of human hair proteins and identification of phosphorylated species. Biol Pharm Bulletin. 2002; 25(5): 569-572.

18. Ramadan MF, Kroh LW, Morsel JT. Radical scavenging activity of black Cumin (Nigella sativa L), Coriander (Coriandrum sativum L), and Niger (Guizotia abyssinica Cass.) crude seed oils and oil fractions. J Agric Food Chem. 2003; 51: 6961–6969.

19. Tran TH, Ramasamy T, Cho HJ, Kim YI, Poudel BK, Choi HG, Yong CS, Kim JO. Formulation and optimization of raloxifene-loaded solid lipid nanoparticles to enhance oral bioavailability. J Nanosci Nanotechnol. 2014; 14(7): 4820–4831.

20. Atkinson AC, Donev AN. Optimum experimental designs. Oxford: Oxford University Press, 1992; 132–189.

21. Wu CC, Chen DH. Facile green synthesis of gold nanoparticles with gum arabic as a stabilizing agent and reducing agent. Gold Bulletin 2010; 43(4): 234–240.