Formulation of Solid Lipid Nanoparticles Containing Hibiscus rosa-sinensis (L.) Extract
Dr. Yogesh S. Thorat1, Parikshit D. Shirure1*, Dr. A. H. Hosmani2
1P G Department of Pharmaceutics, D.S.T.S. Mandal’s College of Pharmacy,
Solapur- 413004 Maharashtra. India.
Department of Pharmaceutics, Government College of Pharmacy, Ratnagiri- 415612 Maharashtra, India.
*Corresponding Author E-mail: pavanshirure105@gmail.com
ABSTRACT:
Hibiscus rosa-sinensis is a genus of flowering plants in the mallow family Malvaceae. It is known as Jaswandi or gudhal in Maharashtra, India. The recognized Indian systems of Medicine are Ayurveda, Siddha and Unani, which use herbs and minerals in the formulations. India which use 15 agro-climatic zones, 4700 plant species of which 15000 are reported to have medicinal properties varying degrees. Hibiscus rosa-sinensis flower used for treatment of many ailments including constipation stomach upset, hair fall, CNS disorder like depression. extract of Hibiscus rosa-sinensis flowers contain vitamins, flavonoids, ascorbic acid, niacin, riboflavin, thiamine and cyaniding diglucoside. Quercetin-3-diglucoside, cyanidin-3-sophoroside-5-glycosides, 3,7-diglucoside, cyanidin-3, 5-diglucoside have been isolated from deep yellow flowers. Flowers of Hibiscus rosa-sinensis can also be made into a kind of pickle or used as a purple dye for colouring foods such as preserved fruit and cooked vegetables. Solid lipid nanoparticles (SLNs) are considered to be the most effective lipid based colloidal carriers, introduced in early nineties. This study was aimed to formulating the hibiscus rosa-sinensis extract loaded solid lipid nanoparticles using Cow’s ghee as lipid core. In the present research work Solid lipid nanoparticles of extract hibiscus rosa-sinensis were prepared high speed homogenization method. The SLN sparticle size analysis depicts that the SLN swere ranging from 222nm to 3264.7nm. These SLN sprepared cow ghee shows Heterogeneous distribution and. SLNs shows maximum entrapment 81.14%. The optimized batch gives 99.21% release in phosphate buffer. It also observed in the physical examination of all batches showed better stability at room temperature.
KEYWORDS: Hibiscus rosa-sinensis extract, Solid lipid nanoparticles, Cow ghee etc.
INTRODUCTION:
Hibiscus rosa sinensis one such plant and belongs to the family Malvaceae. It is a species of tropical Hibiscus in the Hibisceae tribe. It is considered native to East Asia. Although the plant is not related to the true roses, the term ‘Rosa sinensis’ literally means ‘rose of China’ in Latin. It was first named by Carolus Linnaeus (Oguntoye, 2014). It is used for treating loss of appetite, colds, heart and nerve diseases, upper respiratory tract pain and inflammation, stomach irritation, fluid retention and also some disorders of circulation. The accepted Indian systems of medicine are Ayurveda, Unani and Siddha, which uses herbs and natural resources in the formulations. Different extract of Hibiscus rosa-sinensis flowers contain vitamins, flavonoids, ascorbic acid, niacin, riboflavin, thiamine andcyanidingdiglucoside. Quercetin-3-diglucoside, cyanidin-3-sophoroside-5-glycosides, 3, 7-diglucoside, cyanidin-3, 5-diglucoside have been isolated from deep yellow flowers H. Rosa-sinensis are reported to possess cardio-protective, hypertensive, anti-diabetic, anti-convulsant and antioxidant activity in constipation and diarrhea. The leaves and flowers are observed to be promoters of hair growth and aid in healing of ulcers. It has been found to be effective in the treatment of arterial hypertension and to have significant antifertility effect. Flowers of Hibiscus rosa-sinensis can also be made into a kind of pickle or used as a purple dye for colouring foods such as preserved fruit and cooked vegetables.
Solid lipid nanoparticles (SLNs) are considered to be the most effective lipid based colloidal carriers, introduced in early nineties. This is the one of the most popular approaches to improve the oral bioavailability of the poorly water soluble drugs. SLNs are in the submicron size range of 50-1000 nm and are composed of physiologically tolerated lipid components which are in solid state at room temperature. They have many advantages such as good biocompatibility, non toxic, stable against coalescence, drug leakage, hydrolysis, biodegradable, physically stable and good carrier for lipophillic drugs.
The aim of this research was to study the extract of Hibiscus rosa-sinensis (L.) and formulation of solid lipid nanoparticles using Cow ghee as Lipid core.
MATERIALS AND METHODS:
Materials:
Hibiscus rosa sinensis flower were obtained from the local market from solapur. Cow’s ghee as Lipid core. All other ingradients used i.e Pluronic F68, polyethylene glycol 400, tween 80 were of pharmaceutical grade.
Preparation of Extracts of Hibiscus Rosa sinensis by maceration process: (3)
The methanolic extract was obtained by maceration of fresh sepal-less flowers of H. rosa-sinensis for 72 h, followed by filtration and concentrated to remove methanol. The extracted by a slight modification of method described earlier by Harborne.Fresh sepal-less flowers (200 g) of H. rosa-sinensis were macerated in 2 L methanol: 2M HCl (85:15 v/v) solution for 72 h. The extract was then concentrated to 500 ml and filtered. To the filtrate, 100 ml concentrated HCl was added. Mixture was heated in round bottom flask under reflux for 2h. The mixture was then refrigerated until crystals were separated out were then filtered, air dried and then weight extract was recorded.
Fig. 1: Extraction of Hibiscus rosa sinensis
Formulation of Solid lipid nanoparticles for extract of Hibiscus rosa-sinensis:
Hibiscus rosa-sinensis extract loaded solid lipid nanoparticles (SLNs) was prepared by High speed homogenization method. In that disoving drug in Cow ghee in a beaker melted for 60-70ºc then solution was sonicated for 5 min. then in another beaker surfactant and polymers was mixed and this mixture was added to distilled water and stirred using magnetic stirrer for 5 min and then prepared oil phase was added slowly to the aqueous phase then mixture was homogenized for 10 min. while the mixtures was cooled at room temperature SLNs formed.
Fig. 2: Calibration curve of Hibiscus rosa sinensis extract
Fig. 3: TLC for Hibiscus rosa sinensis extract
Fig. 4: Drug entrapment efficiency
Fig. 5 % Drug release optimized batch
Fig.6 Particle size analysis F2 batch
Fig.7 Particle size analysis F3 batch
Fig.8 Particle size analysis F4 batch
Fig.9 particle size analysis F7 batch
Evaluation of solid lipid nanoparticles
1. Drug Entrapment efficiency:(8)
Entrapment efficiency of hibiscus rosa sinensis extract loaded solid lipid nanoparticles by measuring the concentration of unentrapped drug in aqueous medium by centrifugation method the nanoparticles were centrifuged in Remi cooling centrifuge using centrifuge tubes at 5000 rpm for 15min at 4ºc.the supernatant was separated out was determined at 291.5 nm using UV visible spectrophotometer.
The entrapment efficiency was then calculated using following equation.
Total drug content- Free drug
% EE = --------------------------------------------- X 100
Total drug content
Table No. 1 Formulation table
|
Ingradients |
F1 |
F2 |
F3 |
F4 |
F5 |
F6 |
F7 |
F8 |
F9 |
|
Drug (mg) |
100 |
100 |
100 |
100 |
100 |
100 |
100 |
100 |
100 |
|
Cow ghee(ml) |
10 |
10 |
10 |
15 |
15 |
15 |
20 |
20 |
20 |
|
PEG-400(ml) |
10 |
10 |
10 |
10 |
10 |
10 |
10 |
10 |
10 |
|
Pluronic F60(mg) |
100 |
100 |
100 |
100 |
100 |
100 |
100 |
100 |
100 |
|
Tween 80(ml) |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
Distilled water (ml) |
29 |
29 |
29 |
24 |
24 |
24 |
19 |
19 |
19 |
Table no. 2 Characteristics of Hibiscus rosa sinensis extract
|
Sr. No. |
Charecterestics |
|
Solubility |
Soluble in water, methanol and ethanol |
|
Melting Point determination |
180-185ºc |
Table no.3 Drug entrapment efficiency
|
Batches |
EE% |
|
F1 |
48.57% |
|
F2 |
42..85% |
|
F3 |
81.14% |
|
F4 |
53.14% |
|
F5 |
68.00% |
|
F6 |
37.71% |
|
F7 |
62.85% |
|
F8 |
57.71% |
|
F9 |
50.85% |
Table no .4 % drug release optimized batches
|
Time |
Batches |
||
|
F2 |
F3 |
F4 |
|
|
0 |
0 |
0 |
0 |
|
30 |
44.35% |
52.71% |
47.57% |
|
60 |
51% |
72.64% |
65.12% |
|
90 |
67.71% |
79.07% |
71.57% |
|
120 |
74.35% |
91.71% |
78.42% |
|
180 |
80.35% |
101.35% |
87.42% |
|
240 |
88.07% |
115.92% |
99.21% |
2. In-vitro drug release study of SLNs:
Studies of the drug release/diffusion from solid lipid nanoparticles system are directed toward In vitro diffusion studies were carried out using Franz diffusion cell apparatus with a diameter of 25mm and a diffusion area of 3.07cm2. Regenerated cellulose acetate membrane (thickness of 60–65 μm and 0.45μm pore size) was sandwiched between the lower cell reservoir and the glass cell top containing the sample and secured in place with a pinch clamp. Area of 3.07 cm2. Regenerated cellulose acetate membrane (thickness of 60–65μm and 0.45μm pore size) was sandwiched between the lower cell reservoir and the glass cell top containing the sample and secured in place with a pinch clamp. The receiving compartment (volume 15 was filled with pH7.4 phosphate buffer containing 0.5% tween-80 (to maintain sink condition). The system was maintained at 37±0.5 °C by magnetic heater, resulting in a membrane-surface temperature of 32 °C. A Teflon TM coated magnetic bar continuously stirred the receiving medium to avoid diffusion layer effects. A sample was placed evenly on the surface of the membrane in the donor compartment. 2.5ml of receptor fluid were withdrawn from the receiving compartment at scheduled intervals and replaced with 2.5ml of fresh solution. Samples were assayed spectrophotometrically for drug content at 291.5nm.
3. Measurement of particle size:
Particle size analysis was done by Beckman Coulter size analyzer depicts the particle size distribution of optimized SLNs batches. Particle size analysis depict that the SLNs were ranging from 222nm to 3264.7nm. The SLNs prepared with cow ghee shows heterogeneous distribution and shows very irregular pattern except F2 batch which shows smaller particle size as compared the other batches.
The batch F2 shows smaller particle size of 222nm and having polydispersity index (PDI) of 0.329 and diffusion constant 2.221e-008 cm2/sec
RESULTS AND DISCUSSION:
Thin layer chromatography method was used for confirmatory analysis of extracted Hibiscus rosa-sinensis. Stationary phase used was Silicagel Gand mobile phase used was Butanol: ethylacetate: water (10:10:4). Standard and sample are applied on plate and mobile phase was allowed to saturate. Mobile phase is allowed Torun over TLC plate and Rf value was calculated. Standard Rf value for Hibiscus Rosa-sinensis 0.54
Standard Rf values: 0.54
Observed Rf values: 0.79
Charecterization of drug
Standard calibration curve of Hibiscus rosa-sinensis
Calibration curve for Hibiscus rosa-sinensis was found to be linear over concentration range 20-100 µg/ml with equation y= 0.007 x + 0.050 and correlation coefficient 0.992 at 291.5nm. Hence the curve shows a good linearity.
CONCLUSION:
Finally, the study demonstrated that extract of Hibiscus rosa-sinensis can be suitably formulated Solid lipid nanoparticles using Cow ghee as lipid core. SLN produces satisfactory results.
ACKNOWLEDGEMENT:
Authors are very much thankful to Dr. Y.S. Thorat, Associate Professor, D.S.T.S. Mandal’s Collage of Pharmacy, Solapur, Maharashtra, India for his continuous support, guidance and encouragement.
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Received on 23.08.2020 Modified on 24.09.2020
Accepted on 05.10.2020 ©A&V Publications All right reserved
Res. J. Pharma. Dosage Forms and Tech.2021; 13(1):7-11.
DOI: 10.5958/0975-4377.2021.00002.1