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Mokhtar, Wong Tin Wui* Particle Design Research Group, Faculty of Pharmacy Non-Destructive Biomedical and Pharmaceutical Research Centre Universiti Teknologi MARA, 40450 Shah Alam, Selangor, Malaysia. * Introduction Diabetes mellitus is a metabolic disorder characterized by high blood glucose level as a result of pancreatic failure to secrete insulin sufficiently or the bodily resistance towards the action of insulin. The syndrome causes various complications namely chronic renal failure, retinal damage leading to blindness, nerve damage, microvascular damage, impotence, poor wound healing leading to ulceration, gangrene and many others.



Fig. 1: Clinical complications of diabetes mellitus


The prevalence of diabetes mellitus in Malaysia was escalating at a phenomenal scale over the years. Over 90% of the diabetic patients suffer from type II diabetes (insulin resistance or reduced insulin sensitivity, in combination with reduced insulin secretion) while the remaining 10% of patients have type I diabetes (loss of the insulin-producing beta cells of the islets of Langerhans in pancreas, leading to a deficiency of insulin). Though both are pathophysiologically different, they share common morbidity and mortality. Lowering the blood glucose level is the primary step to prevent and reduce various diabetic complications of diabetic patients. Medically, patients with type I diabetes require the administration of exogenous insulin. To date, the insulin is administered by means of subcutaneous route. The delivery of insulin through injection induces pain and thus has a low level of patient compliance. One of the alternative routes of insulin administration is by means of oral delivery. Nonetheless, the development of oral dosage form carrying the insulin has encountered immense difficulty with respect to proteolysis of insulin in the gastrointestinal tract and poor permeability of insulin molecules across the intestinal epithelium.


Poor permeability of insulin through epithelia Enzymatic activity in gastrointestinal tract that could proteolyses insulin

Fig. 2: Physiology of gastrointestinal tract

The invention of nanoparticles has led to the formation of a new direction in design of oral insulin delivery system. Nanoparticles can be fabricated using polysaccharides. Polysaccharides are found in abundance, inexpensive and available in a variety of

structures with a variety of physicochemical properties. Polysaccharides can be easily modified chemically and biochemically and are highly stable, safe, non-toxic, hydrophilic and gel forming. Pectin is an anionic polysaccharide present in the cell wall of most plants. It is made of (1-4)-linked D-galacturonic acid residues interrupted by 1,2-linked L-rhamnose residues. The pectin is potentially useful for making nanoparticles via crosslinking reaction with cationic agents. Utilizing pectin as matrix polymer, present study aims to investigate the effects of standing time of pectin insulin solution on size, zeta and insulin association efficiency of the formed pectin nanoparticles. Objective The present study investigated the effects of standing time of pectin-insulin solution on the association efficiency of the formed nanoparticles. Experimental The pectin-insulin nanoparticles were prepared through subjecting the solution mixture of pectin and insulin to varying lengths of standing time at pH 2.5 and 4C, prior to ionotropic gelation with zinc chloride solution. The formed nanoparticles were subjected to size and zeta potential analysis using dynamic light scattering and electrophoretic light scattering techniques respectively at 25C in triplicates (Malvern, UK).The insulin association efficiency, defined as the quotient of amount of encapsulated insulin to theoretical amount of insulin employed in the preparation of nanoparticles, in the unit of percentage was analyzed by means of high performance liquid chromatography (Agilent, USA). Results and discussion Fig. 3: Size, zeta potential and insulin association efficiency of nanoparticles prepared through keeping the pectin-insulin solution at various standing time prior crosslinking. a)
400 350 -5 300 250


0 0 24 48 60

Zeta potential
0 24 48 60



200 150 100 50 0




Standing time

Standing time





Insulin AE%




0 0 24 48 60

Standing time

The freshly prepared nanoparticles had average size, zeta potential and insulin association efficiency of 283 nm, -11.4 mV and 39.1 % respectively (Fig 3). Standing of pectininsulin solution for 24, 48 and 60 hours did not affect the size of nanoparticles formed substantially (Fig 3). Standing of pectin-insulin solution for 24 hours did not result in significant changes in the association efficiency of insulin in nanoparticles (Fig 3). The insulin association efficiency of nanoparticles was increased by 10 % upon standing the pectin-insulin solution for 48 hours (Fig 3). Apparently, higher association efficiency in the nanoparticles was accompanied by a higher magnitude of negative zeta potential (Fig 3). This could probably associated to hindrance of pectin-Zn interaction by the embedded insulin molecules. Prolonged standing of pectin-insulin solution for 60 hours brought about a decrease in insulin association efficiency to 38.2 % probably due to insulin degradation and/or desorption (Fig 3). Conclusion The insulin association efficiency of pectin-insulin nanoparticles can be promoted through standing the pectin-insulin solution for more than 24 but less than 60 hours. References Allemann, E., Leroux, J.C., Gurny, R., 1998. Polymeric nano- and microparticles for oral delivery of peptides and peptidomimetics. Advanced Drug Delivery Reviews. 34, 171189. Cheng, K., Lim, L.V. 2004. Insulin-loaded calcium pectinate nanoparticles: effects of pectin molecular weight and formulation pH. Drug Dev. Ind. Pharm. 30,359-367.