Formulation Development and Evaluation of Nasal In- Situ Gel of Hydrocortisone

 

Pradnya M Khandagale¹, Manisha M. Rokade², Dipti G. Phadtare³

¹Department of Quality Assurance Techniques, R. G. Sapkal Institute of Pharmacy, Anjaneri, Nashik-422213, Maharashtra, India.

²Department of Quality Assurance Techniques, R. G. Sapkal Institute of Pharmacy, Anjaneri, Nashik-422213, Maharashtra, India.

³Department of Pharmaceutical Chemistry, R.G. Sapkal College of Pharmacy, Anjaneri, Nashik-     422213, Maharashtra, India.

 

ABSTRACT:

Over a few decades, advances in the in-situ gel technologies have spurred development in may medical and biomedical applications including controlled drug delivery. For locally acting intra nasal drugs, an extended residence time in the nasal cavity is desirable and related to a prolonged effect. The aim of the present work was to design a nasal delivery system with improved mucoadhesive properties that could provide prolonged retention time for the treatment of the allergic rhinitis. A 3² factorial design was used to investigate the effect of amount of Poloxamer 188 and carbopol 934as an independent variables. % drug release and mucoadhesive strength were taken as dependant variables. The formulations were tested for gelation study, viscosity study, gel strength, mucoadhesion study, drug content and stability study. Gelation was determined by physical appearance, viscosity study of solution and gel formulations indicated that increase in polymer concentration increase the viscosity. Gel strength was found in the range of 0.55-2.36 sec. The mucoadhesive force in terms of detachment stress increase with increase in the concentration of carbopol 934.  A stability study for optimized formulation as per ICH guideline for 90 days showed no change in pH, drug content and viscosity. The developed in-situ gelling system for Hydrocortisone using poloxamer188 in combination with carbopol 934 with improved mucoadhesive properties that could provide prolonged retention time for the tratement of allergic rhinitis.

 

 

 

 

 

INTRODUCTION:

Nasal therapy is the recognised form of treatment in Ayurvedic systems of Indian medicine. It is also called as nasaya karma. The nasal mucosa itself and the drug delivery systems affect drug absorption through the nasal route, and is invaluable. A stable, safe and effective nasal product can be developed through appropriate and adequate preformulation studies of drug. Nasal drug delivery system provides excess of easy application of drug with the possibility of self administration by removing the chance of unwanted painful condition associated with injection form of drug delivery.^(1,2) The greater permeability of nasal mucosa with large surface area affords a rapid onset of therapeutic effect. The appealing advantage of nasal drug delivery is the possibility of targeting central nervous system(CNS) by passing blood brain barrier(BBB). The drug absorbed nasally via. olfactory epithelium are reported to enter in the olfactory neurons and supporting cells and subsequently into the brain, which reduced not only the systemic toxicity of centrally acting drugs but also enhanced therapeutic efficacy. Nasal route is the route of choice for rapid mass immunization in developing countries. Despite having large number of advantages, bioavailability of nasal dosage form is hindered. poloxamer 188 is generally occur as white, free flowing prilled granules, or as cast solids. They are practically odourless and tasteless. Poloxamers are stable materials and their aqueous solutions are stable in the presence of acids, alkalis and metal ions. However, aqueous solutions support mold growth. It contain less than 0.5% w/w water and are hygroscopic only at relative humidity greater than 80%. Poloxamers are used in a variety of oral, parenteral and topical pharmaceutical formulations and are generally regarded as nontoxic and non-irritant materials. Poloxamers are not metabolized in the body.⁽³⁾

Carbopol is white colored, Fluffy, acidic, hygroscopic powders with a slight characteristics odor. Carbomer range are synthetic high molecular weight crosslinked polymers of acrylic acid, which confirm to USP/INF. They are water soluble polymer use to thicken to produce wide range of viscosities and flow properties. It is suspend soluble ingredient and it will stabilize and emulsify emulsion. The decomposition of carbopol 934 occurs within 30 min at 260°C. It is Mucoadhesive, emulsifying agent, release modifying agent, suspending agent, tablet binder, viscosity increasing agent.⁽³⁾

 

Hydrocortisone is the pharmaceutical term for cortisol used in oral administration, intravenous injection, or topical application. It is used as an immunosuppressive drug, given by injection in the treatment of severe allergic reactions such as anaphylaxis and angioedema, in place of prednisolone in patients needing steroid treatment but unable to take oral medication, and perioperatively in patients on long-term steroid treatment to prevent Addisonian crisis. It may also be injected into inflamed joints resulting from diseases such as gout. It may be used topically for allergic rashes, eczema, psoriasis, itching and other inflammatory skin conditions. Topical hydrocortisone creams and ointments are available in most countries without prescription in strengths ranging from 0.05% to 2.5% (depending on local regulations) with stronger forms available by prescription only. Covering the skin after application increases the absorption and effect. Such enhancement is sometimes prescribed, but otherwise should be avoided to prevent overdose and systemic impact.⁽⁴⁾

An attempt was made to develop the temperature sensitive nasal In-situ gel Hydrocortisone (2%) for controlling the drug release in the nasal tissues. Two polymers have been used i.e. Poloxamer 188 and carbopol 934. Poloxamer 188 is Temperature sensitive and act as a gelling agent and carbopol 934 is the pH sensitive mucoadhesive polymer.⁽⁵⁾

 

MATERIALS AND METHODS:

Poloxamer 188 was purchase from BASF chemicals, Mumbai, and Carbopol 934 from Loba Chemie, Pvt.Ltd. India.

Method

Gel was prepared by cold technique method. The Poloxamer 188 and Carbopol 934 was dissolved completely in distilled water and allow to hydrate for overnight. The 2% of drug solution was added in distilled water and Propylene glycol 0.1% v/v. then Benzalkonium chloride was added as a preservative in the above solution. On next day both the solution were mixed together with continues stirring under cold condition. This resultanting formulation was kept overnight in a refrigerator until clear solution was obtained. ⁽⁵⁾

 

Preformulation study of drug ^(5,6)

Preformulation can be described as the step that deals with the acquisition of data on the drug compound and its excipients, which are then analyzed or processed into information, and ultimately transformed into knowledge in the form of a recommended formulation. The area of preliminary- preformulation represents the scope of activities that facilitate the design of preformulation studies and is often theoretical in nature. Preformulation testing is the first step in rational development of dosage forms of a drug substance. It can be defined as an investigation of physical and chemical properties of a drug substance alone and when combined with excipients. The overall objective of preformulation testing is to generate information useful to the formulator in developing stable and bioavailable dosage forms that can be mass produced.

 

Description

Drug sample was evaluated for color and texture.

Melting Point

Melting point of Hydrocortisone was determined by taking a small amount of sample in a capillary tube closed at one end and placed in melting point apparatus. The melting point was noted in triplicate.

 

Solubility

The solubility of Hydrocortisone was checked in different solvents like distilled water, Hcl (0.1 N), methanol, NaOH, Acetone, phosphate buffer (pH 6.8 and 7.4).

 

Ultraviolet -Visible Spectroscopy ^(7,8)

Determination of λmax in Methanol

The UV spectrum of Hydrocortisone was obtained using Ultraviolet Spectrophotometer (shimadzu 1800 series). Accurately weighed 10 mg of the drug was dissolved in sufficient quantity of Methanol and volume was made up to 100 mL to obtain a concentration of 100μg/mL. 1 ml of aliquot was withdrawn and volume was made up to 10 ml using Methanol to obtain the concentration of 10 μg/ml. The resultant solutions scanned from 200- 400 nm and spectrum was recorded to obtain the value of maximum wavelength.

 

Determination of  λmax in phosphate buffer pH 6.8

Hydrocortisone (10mg) was weighed accurately and transferred to 100 ml volumetric flask. It was then dissolved in little quantity of Methanol and diluted up to 100 ml with phosphate buffer pH 6.8. The stock solution of 100μg/ml was used to prepare different dilutions in the range of 2-20μg/ml. The absorbance of resulting solutions were measured at 239 nm using phosphate buffer pH 6.8 as a blank by UV-visible spectrophotometer.

 

Fourier transform Infrared spectroscopy (FTIR)⁽⁹⁾             

The FTIR spectra of pure drug and physical mixture (Drug + Poloxamer 188 + Carbopol 934) was carried out. The physical mixture were prepared and sample kept for 1 month at 40°C. The infrared absorption spectrum of drug and physical mixture of drug and polymers was recorded with the wave number 4000- 400 cm^-1.

The quantities of drug and other ingredients were weighed and formulation were prepared in following manner:

 

Cleaning of glassware and container:

All the glassware’s were washed with distilled water and then sterilized by drying at 160°c for 1 hr in hot air oven.

 

Preparation of solution’A’:

Accurately weighed quantity (0.2gm) of Hydrocortisone and was dissolved in Isopropyl alcohol.

 

Preparation of polymer dispersion ‘B’:

Poloxamer188 and carbopol was dissolved in water under cold condition with continuous stirring. And the solution was kept to hydrate for 12 hr to produce a clear solution.

 

Mixing:

After 12hrs both the solution A and B were mixed together under cold condition with continuous stirring.

 

Aseptic filling to container:

The formulation was aseptically transferred to previously sterilized glass vials and sealed.

 

Formulation optimization:

3² full factorial design was applied to the formulation that showed the satisfactory results. To see the effects of conc. of variables Poloxamer 188 and Carbopol 934 on various responses like % drug release. composition of all batches is shown in Table no. 1

 

 

 

Table No. 1: Composition of formulation batches as per 3²facorial design.

Formulation code Ingredients	F1	F2	F3	F4	F5	F6	F7	F8	F9
Hydrocortisone (% w/v)	0.2	0.2	0.2	0.2	0.2	0.2	0.2	0.2	0.2
Poloxamer-188 (%w/v)	14	16	18	14	16	18	14	16	18
Carbopol-934 (%w/v)	0.1	0.1	0.1	0.15	0.15	0.15	0.2	0.2	0.2
Propylene Glycol (%w/v)	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.01
Benzalkonium Chloride (%w/v)	1	1	1	1	1	1	1	1	1
Triethanolamine	q.s.	q.s.	q.s.	q.s.	q.s.	q.s.	q.s.	q.s.	q.s.
Purified water (ml)	100	100	100	100	100	100	100	100	100
Iso Propyl alcohol (ml)	5	5	5	5	5	5	5	5	5

 

 

Evaluation of Gel⁽¹⁰⁾

Clarity

The clarity of the gel was done by visual inspection under black and white background.

 

pH

pH of each formulation was determined by using Digital pH meter( sistronic Digital pH meter 335) was previously calibrated by pH 4 and pH 7. The pH values were recorded immediately after preparation.

Rheological study 

Viscosity⁽¹¹⁾

The rheological properties of gels were determined by the Brookfield Viscometer; type DV-II+ PRO using spindle no. LV 3(63). Viscosities of the formulations were taken at two different temperatures i. e. at Room temperature and at 37°c with varying shear rate.

 

 

 

Measurement of Gelling capacity

The gelling capacity of the formulations were determined by placing 1 drop of the prepared formulation into a vial containing 2 ml of freshly prepared SNF solution. Gelation was assessed visually and noting the time for the gelation and the time taken for the gel formed to dissolved.

 

Gel Strength

A sample of 25 ml of gel was put in a 50 ml graduated cylinder. A weight of 14.33 gm was placed on the gel surface. The gel strength which is an indication for the gel at physiological temperature was determined by the time in seconds required by the weight to penetrate 5 cm into the gel. All measurement were performed in triplicate.

 

Figure No.1:  Gel Strength measuring Device

(A) Weights (B) Device (C) Graduated Cylinder (D) Gel

 

Mucoadhesive Strength

“Detachment Stress is the force required to detach the two surfaces of mucosa when a formulation/gel is placed in between them”. The detachment stress was measured by using a modified analytical balance.

 

i) Fabrication of equipment: The equipment was fabricated by us in the laboratory as shown in figure 3. A double beam physical balance was taken, both the pans were removed. The left pan was replaced with a brass wire, to which was hanged a teflon disc (A), also locally fabricated. The dimensions are 2 cm height and include an expanded cap of diameter 3.8 cm and thickness 2 cm. Another teflon disc of 2 cm height and 1.5 cm diameter was placed right below the suspended disc upon the base of the balance. The right pan (C) was replaced with a lighter pan so that, the left pan weighs 5.25 gm more than the right pan. The lower polypropylene block was intended to hold the mucosal tissue (D) of goat nasal mucosa and to be placed in a beaker containing simulated nasal solution pH 6.7 (E). 

 

ii) Measurement of adhesion force: 

The following procedure was used for all the test formulations using the above equipment. The nasal mucosa was removed from refrigerator and allowed to attain equilibrium with ambient conditions in the laboratory. The goat nasal mucosa was carefully excised, without removing connective and adipose tissue and washed with simulated nasal solution. The tissue was stored in fresh simulated nasal solution. Immediately afterwards the membrane was placed over the surface of lower teflon cylinder (B) and secured. This assembly was placed into beaker containing simulated nasal solution pH 6.7 at 37 ± 2°C.From each batch, some quantity of gel was taken and applied on the lower surface of the upper teflon cylinder. The beaker containing mucosal tissue secured upon lower cylinder (B), was manipulated over the base of the balance so that, the mucosal tissue is exactly below the upper cylinder (A). The exposed part of the gel was wetted with a drop of simulated nasal solution, and then a weight of 10 gms was placed above the expanded cap, left for 10 minutes. After which the gel binds with mucin. The weight was removed. Then slowly and gradually weights were added on the right side pan till the gel separates from the mucosal surface/ membrane.

The weight required for complete detachment is noted (W1) (W1-5.25G)) gives force required for detachment expressed in weight in grams. Procedure was repeated for two more times. Average was computed and recorded. 

 

iii) Calibration of test equipment:  

Initially, a gel from the same batch was taken ten times and individual force required for complete detachment was noted and S. D.was calculated. 

 

iv) Force of adhesion (N)

= (bioadhesive strength/1000) × 9.81

 

Bond strength (N/m²)

= force of adhesion (N)/surface area of disk (m²)

 

Figure No. 2 : Modified Balance for Mucoadhesive Study

A: Modified balance, B: Weighing pan, C: Weight D: Gel, E: Nasal mucosa F: Polypropylene cylinder.

Drug content⁽¹²⁾

1 ml of the formulation was added in 10ml volumetric flask and make up volume upto 10ml with phosphate buffer pH 6.8. from this solution remove 1ml solution and make up volume upto 10ml with methanol. And at the fixed wavelength the absorbance of the formulation was carried out.

 

In-vitro Drug release study⁽¹³⁾

In-vitro release study of the formulated in-situ gel was carried out by using diffusion cell through egg membrane as a biological membrane. Diffusion cell with inner diameter 1.4cm was used for the study. The formulation 1 ml were placed in donor compartment and Freshly prepared 100 ml simulated nasal electrolyte solution (sodium chloride 0.745gm, potassium chloride 0.129 gm, calcium chloride dehydrated 0.005gm, distilled water q.s. 100ml) in receptor compartment. Egg membranes were mounted in between donor and receptor compartment. The position of the donor compartment was adjusted so that egg membrane just touches the diffusion medium. The whole assembly was placed on the thermostatically controlled magnetic stirrer. The temperature of the medium was maintained at 37°C ± 0.5°C. 2ml of sample is withdrawn from receiver compartment after 30 min, 1, 2, 3, 4, 5, 6, 7 and 8 hrs and same volume of fresh medium is replaced. The withdrawn samples was diluted to 10ml in a volumetric flask with Methanol and analyzed by UV spectrophotometer at 239 nm.

 

 

Figure No.3: Laboratory designed diffusion cell.

 

In-vitro drug permeation study

Natural membranes are utilized to determine in vitro permeation study to mimic the in vivo permeation patterns. In this experiment goat mucosa was utilized because the respiratory area of the goat is large and it is easy to get. Fresh mucosa tissue wa removed from the nasal cavity of goat. The tissue was placed on the diffusion cell with permeation area 0.786 cm^-2. The acceptor chamber of the diffusion cell with a volume capacity 100ml was filled with simulated nasal fluid contain accurately 7.45 mg/ml NaCl, 1.29mg/ml KCl and 0.32mg/ml CaCl₂.2H₂O. 0.5ml (10mg) of formulation was placed in donor compartment. At predetermined time intervals of 30 min, 1,2,3,4,5,6,7, and 8 hrs 1ml of sample was withdrawn from the acceptor compartment replacing the sample removed with SNF after each sampling for period of 8hrs. then samples are specifically diluted and absorbance was noted at 239nm permeability coefficient (p) was calculated by the following formula:

 

P =(dQ/dt) / ( C₀ × A)

 

Where, dQ/dt is the flux or permeability rate (mg/h), C₀ is the initial concentration in the donor compartment, and A is the effective surface area of nasal mucosa.

 

Stability studies ⁽¹⁴⁾

 

Table No.2: Test conditions for stability study

Test Conditions
Duration of study:	3 months
Temperature conditions:	Room temperature 250C±20C
Relative humidity conditions:	75%± 5%
Frequency of testing the samples:	30 Days

 

The formulations were evaluated mainly for their physical characteristics at the predetermined intervals of 30 Days like appearance/clarity, pH, viscosity and drug content.

 

RESULT AND DISCUSSION:

Preformulation study-

Description-

It is White or almost white crystalline powder with the description that found in literature survey.

 

Melting Point-

Melting point of powder drug was shown in table no. 3

Melting Point (°C)
Literature	Practical
272-273°C	273-274°C

 

Solubility-

Hydrocortisone is soluble in water. Practically soluble in Methanol, Ethanol. Isopropyl alcohol

 

Ultraviolet-Visible Spectroscopy Study

Determination of λ_max in Methanol

 

Figure No.4: UV Spectra of Hydrocortisone in Methanol

The UV spectrum of Hydrocortisone (10μg/mL) exhibited wavelength of absorbance maximum at 239 nm. This is near to the reported value. However, keeping in mind the probable concentrations likely to be encountered while carrying out In-vitro release studies and considering the predicted theoretical λmax involved, the working λmax was decided as 239 nm.

 

Calibration of Hydrocortisone in Methanol

The calibration curve was found to be linear in a concentration range of 5-30 µg/ml having coefficient of regression value R²= 0.9991 at 238nm is shown in Figure No.3

 

Table No.3.  Absorbance of Hydrocortisone in Methanol

Sr. No.	Concentration  (ppm)	Absorbance
1.	5	0.3451
2.	10	0.5384
3.	15	0.7483
4.	20	0.9878
5.	25	1.2104
6.	30	1.4277

 

 

 

Figure No.5: Calibration curve of Hydrocortisone in Methanol

 

Infrared Spectrum

The absorption bands shown by Hydrocortisone are characteristics of the groups present in its molecular structure. The presence of absorption bands corresponding to the functional groups present in the Hydrocortisone confirms the identification and purity of drug.

 

Figure No. 6: FTIR Spectrum of Hydrocortisone

 

Figure No. 7: FTIR Spetrum of Hydrocortisone with polymers

 

Evaluation of Nasal In-situ gel

Clarity-

All the formulations are found to be clear solution against a dark and white background.

 

 

 

 

 

 

 

pH-

pH of all the formulations were shown in the Table No. 4

Sr. No	Formulation code	Observed pH (±S.D.)
1	F1	6.3±0.01
2	F2	5.7±0.07
3	F3	6.3±0.02
4	F4	6.2±0.05
5	F5	5.9±0.03
6	F6	6.3±0.007
7	F7	6.1±0.01
8	F8	6.0±0.05
9	F9	6.15±0.1

 

Rheological Study-

Viscosity-

At Room Temperature

 

 

Figure No.8: Viscosity Profile of formulation RT

 

 

Figure No. 9: Viscosity Profile of Formulations at 37^ºC

Gelling Capacity-

Formulation Code	Gelling Capacity
F1	+
F2	+
F3	++
F4	+++
F5	++
F6	+++
F7	++
F8	++
F9	+++

(+: Gel formed after a few minutes, dissolves rapidly, ++: Immediate gelation, remains for few hours, +++: Immediate gelation, remains for extended period.)

 

Gelling strength-

Sr. No	Formulation code	Gel strength (sec)(±S.D.) at RT	Gel strength (sec)(±S.D.) at 37°c
1	F1	0.44±0.007	0.55  ± 0.007
2	F2	0.61 ± 0.007	0.87±0.007
3	F3	0.82 ±0.007	1.3±0.007
4	F4	0.77 ± 0.007	0.98±0.007
5	F5	0.86 ± 0.007	0.97±0.007
6	F6	0.95 ± 0.007	2.05±0.007
7	F7	0.75 ± 0.007	0.89±0.007
8	F8	1.6 ± 0.03	2.34±0.007
9	F9	2.03 ± 0.007	2.36±0.007

 

 

 

 

 

 

 

 

 

 

Mucoadhesive strength

At RT

Formulation code	Detachment Force (N) (±S.D.)	Bond strength (N/m2)(±S.D)
F1	0.04411 ± 0.001	0.00241± 0.005
F2	0.05036± 0.002	0.0027± 0.002
F3	0.05637± 0.005	0.0031± 0.005
F4	0.06394± 0.003	0.0035± 0.001
F5	0.06725± 0.007	0.0037± 0.0001
F6	0.07257± 0.001	0.0040± 0.001
F7	0.07762± 0.001	0.0042± 0.002
F8	0.08010± 0.001	0.0044± 0.002
F9	0.08469± 0.001	0.0046± 0.005

 

At 37°C

Formulation code	Detachment Force (N) (±S.D.)	Bond strength (N/m2)(±S.D)
F1	0.066 ± 0.00031	0.0031± 0.005
F2	0.068± 0.0001	0.0037± 0.001
F3	0.077± 0.005	0.0042± 0.001
F4	0.082± 0.001	0.0045± 0.002
F5	0.084± 0.0007	0.0046± 0.005
F6	0.087± 0.004	0.0048± 0.001
F7	0.090± 0.0003	0.0050± 0.005
F8	0.091± 0.007	0.0089± 0.002
F9	0.092± 0.00031	0.0051± 0.002

 

Drug content

Formulation Code	Drug content (%) (±S.D.)
F1	92.10±0.007
F2	89±0.007
F3	93±0.007
F4	92.10±0.007
F5	98.68±0.007
F6	102±0.007
F7	93±0.007
F8	96.05±0.007
F9	98.68±0.007

 

 

In-vitro drug release

Cumulative Drug Release (%) (±S.D.)
Time in (Hrs.)	F1	F2	F3	F4	F5	F6	F7	F8	F9
0	0	0	0	0	0	0	0	0	0
30 min	4.25± 0.001	6.21± 0.05	10.11± 0.007	3.47± 0.001	8.16± 0.02	23.39± 0.05	8.16± 0.007	23.78± 0.09	23.78± 0.1
1	8.16± 0.04	11.68± 0.019	17.93± 0.05	6.21± 0.005	17.43± 0.001	23.79± 0.005	21.83± 0.007	24.57± 0.1	33.55± 0.09
2	15.97± 0.01	33.55± 0.009	33.55± 0.007	12.07± 0.005	29.65± 0.02	29.65± 0.001	29.26± 0.007	29.65± 0.05	40.98± 0.001
3	27.69± 0.001	45.27± 0.001	45.28± 0.001	27.30± 0.05	32.38± 0.007	35.51± 0.001	39.42± 0.02	35.90± 0.05	49.58± 0.009
4	47.23± 0.009	46.45± 0.007	46.06± 0.034	48.79± 0.04	39.03± 0.5	57± 0.004	55.05± 0.009	56.61± 0.001	70.28± 0.07
5	60.90± 0.001	53.10± 0.001	58.96± 0.007	58.95± 0.001	46.45± 0.09	61.30± 0.05	64.04± 0.01	61.69± 0.001	78.50± 0.2
6	62.80± 0.02	58.18± 0.05	62.09± 0.009	60.13± 0.007	58.57± 0.001	68.30± 0.001	71.07± 0.005	69.90± 0.009	85.54± 0.007
7	66.78± 0.2	68.73± 0.4	66.39± 0.001	65.61± 0.001	67.95± 0.007	80.46± 0.007	72.64± 0.001	82.02± 0.009	96.09± 0.001
8	72.64± 0.05	82.62± 0.04	79.68± 0.009	72.64± 0.001	80.85± 0.001	92.97± 0.004	86.71± 0.007	94.14± 0.004	98.45± 0.001

 

Figure No. 10: In-vitro drug release profile

 

Drug permeation study

Time	F1	F2	F3	F4	F5	F6	F7	F8	F9
30 Min	15.97	16.75	15.58	12.07	8.55	9.33	5.42	8.16	8.94
1hr	40.58	39.41	23.79	14.02	14.41	12.85	7.77	12.46	14.02
2hr	55.82	56.99	49.96	30.43	19.49	20.27	10.11	23.01	26.52
3hr	80.44	81.22	59.73	45.27	46.84	33.16	13.63	37.07	38.24
4hr	85.14	92.17	65.99	64.42	70.67	61.29	23.40	53.09	57.39
5hr	90.218	93.35	74.59	74.58	76.54	73.41	38.24	62.86	78.11
6hr	92.18	95.70	80.85	80.06	80.84	81.62	51.53	78.10	78.49
7hr	95.71	95.27	97.26	95.30	97.65	84.36	64.82	82.41	89.83
8hr	97.28	98.07	98.45	97.27	98.05	94.14	92.71	93.35	98.83

 

 

Figure No. 11: In-vitro drug permeation study profile

 

 

Figure No.12: Surface response plot showing effect of Poloxamer 188 and Carbopol 934 on % drug release

Stability Study

Stability study of optimized F9 batch was shown below

Stability study data for F9 batch

Sr. No	Observation		Before stability testing		During study
					30  Day		30  Day		30 Day
1.	Clarity		Clear		Clear		Clear		Clear
2.	Visual appearance		Transparent		Transparent		Transparent		Transparent
3.	pH		6.15±0.17		6.16±0.01		6.15±0.01		6.15±0.01
4.	Viscosity (rpm)		At RT	At 37°C	At RT	At 37°C	At RT	At 37°C	At  RT	At 37°C
		5	529.9	600.9	530	601	531	602.3	529.5	530
		10	490.1	568.9	489	567.8	490	568.2	489.7	568.4
		15	435.8	535.4	435	534.9	435.9	535.1	436	534.8
		20	430.4	490	430	491.3	429.5	430.5	430.4	431.7
		25	392.5	455.9	391	455.2	392.4	444.6	393.7	445.3
		30	335.2	415.7	335	415	334.5	414.6	335.6	415.5
5.	Drug content		98.68±0.01		98.69±0.014		98.70±0.019		98.67± 0.01

 

Formulations at room temperature were found to be stable up to 3 months. There is no change in drug content, pH, clarity and viscosity.

 

CONCLUSION:

The nasal route of administration for systemic drug delivery offers a number of advantages compared to conventional routes, especially for peptide and protein drugs. Nasal delivery is being increasingly considered to be an alternative route for drugs that currently require parenteral administration to achieve good efficacy, or where circumstances make oral delivery difficult. The nasal route is an alternative for several reasons, including rapid absorption into the systemic circulation, elimination of hepatic first pass metabolism and low proteolytic activity in the nasal mucosa.  From the above investigation, the temperature sensitive in-situ gel of Hydrocortisone (2%) for controlling the drug release in the nasal tissues. Two polymers have been used i. e. Poloxamer 188 which is Temperature sensitive and also act as a gelling agent and Cabopol 934 act as pH sensitive mucoadhesive polymer.  These conditions resemble the physiological conditions of the nose. The Temperature sensitive in-situ nasal gel so prepared were characterized for its clarity, pH, viscosity, gel strength, mucoadhesive strength, drug content, in-vitro drug release and in-vitro permeation.

 

The following conclusion can be drawn from the present study:

1) Preformulation evaluation study has proven the identity and purity of. Hydrocortisone.

2) Infrared spectroscopy studies of Hydrocortisone alone and their physical mixture revealed that, is compatible with all polymers used.

3) All formulations were examined for visual appearance and were found to be transparent.

4) pH of all the formulations was found to be in between the nasal pH range (4.5-6.5)which is in tolerable range in contact with nasal tissues.

5) The viscosities of all the formulations were greatly affected by concentration of Poloxamer-188 and Carbopol 934. Formulation containing Poloxamer-18% and 0.2%w/v Carbopol 934 showed optimum viscosity and exhibited pseudoplastic behaviour.

6) Gel strength and Mucoadhesive strength of formulations resembles to the viscosity results.

7) Drug content of all the formulations was found to be in between 89-98.68% which was in acceptable range.

8) The release kinetics results obtained indicate that formulation containing Poloxamer-188 18% and 0.2%w/v Carbopol 934 showed highest release i.e. 98.45% after 8 hrs which indicates that the formulation have shown prolonged release. This optimized formula was also confirmed by design expert 7.0.0 optimization software.

9) The optimized formulation F9 showed good stability and no change in any physical characteristics over a 3 months period at 25°C±2ºC Temperature and 75%±5% Relative Humidity.

 

Thus Hydrocortisone 2% Temperature sensitive in-situ nasal gel formulation fulfils all necessary parameters required for nasal use. This optimized formulation having improved viscosity and better mucoadhesive property may improve the bioavailability of nasal administration of Hydrocortisone in gel form and can be alternative to the conventionally administered oral formulation. Also, the common problem of food interaction seen with oral administration can be overcome by the use of the novel dosage form developed in this study.

 

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Received on 16.05.2018                Accepted on 15.06.2018               

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