FORMULATION AND EVALUATION OF pH TRIGGER NASAL IN-SITU GEL OF LEVOCETIRIZINE HYDROCHLORIDE FOR LOCALIZED TREATMENT OF ALLERGIC RHINITIS

FORMULATION AND EVALUATION OF pH TRIGGER NASAL IN-SITU GEL OF LEVOCETIRIZINE HYDROCHLORIDE FOR LOCALIZED TREATMENT OF ALLERGIC RHINITIS

Upasana Khatri, Sunil Saini and Meenakshi Bharkatiya ^*

B. N. Institute of Pharmaceutical Sciences, B. N. University, Udaipur, Rajasthan, India.

ABSTRACT: Objective: This dissertation work aimed to prepare Levocetirizine HCL (LCH) nasal in-situ gel, a second-generation antihistamine, to improve its retention time, sustain release, enhance the bioavailability, and avoid first-pass metabolism. Process: The in-situ nasal gel was prepared to consume Carbopol as a gelling agent, Hypromellose (HPMC) as a thickening or thickness-imparting agent, and buffering agents to maintain pH. An assessment was conducted to determine the solubility, pH, rheological characteristics, consistency of content, adhesive properties to mucous membranes, gel strength, in-vitro release characteristics, the kinetics of release date, isotonicity, and sterility of the sample. Results: All formulations had a clear appearance with good gelling capacity, and drug content was found to be >87%. The pH values of the formulations were found to be acceptable, ranging from 6.2 ± 0.10 to 6.4 ± 0.44, and no irritation of the mucosal membranes was observed. The shearing character was found between 200-1400 at pH 6.4 and 2000-5500 cps at pH 7.4 when measured at 20 rpm. They also showed adequate gel strength of 14 ± 0.35 and mucoadhesion values ranging from 854 ± 0.33 dynes/cm2 to 5517 ± 1.88 dynes/cm², as well as sustained drug release. Based on these results, the optimal concentration range for both polymer and stand was determined to be between 0.775 and 0.800 for all parameters. Therefore, it can be concluded that in-situ nasal gel of LCH is a promising drug delivery system that could enhance bioavailability by overcoming first-pass metabolism.

Keywords: Allergic rhinitis, Antihistaminic, Levocetirizine hydrochloride, Nasal obstruction, Mucoadhesive strength, Viscosity, HPMC, Carbopol

INTRODUCTION:

Allergic Rhinitis: Allergic rhinitis (AR) is a prevalent and chronic type 2 inflammatory disorder that is heterogeneous in nature and has been present for a long duration ^{1, 2}. The incidence of AR increased daily, affecting a significant number of individuals worldwide ^{3, 4}.

The pathogenesis of AR is a complex process involving the interaction between environmental allergens, the immune system, and the nasal mucosa. Upon the entry of an allergen into system, is identified as a foreign substance by the immune system, which triggers an immune response ^{5, 6}.

Immunoglobulin E (IgE) antibody production starts automatically when an allergen triggers the immune system, which attaches to the allergen and activates mast cells and basophils. These cells release various mediators, such as histamine and leukotrienes, that cause inflammation and swelling of the nasal mucosa. Consequently, these events result in the symptoms associated with allergic rhinitis ⁷. Various allergens like pollen, dust, animal dander, and mold cause allergic rhinitis (AR). Its symptoms include sneezing, nasal congestion, runny nose, itchiness of the nose, mouth, or throat, and watery eyes. These symptoms can range from mild to severe and may persist for days or weeks ⁸. Diagnosing allergic rhinitis (AR) is typically based on the patient's medical history, symptoms, and physical examination. Sometimes, allergy testing may be conducted to identify the specific allergen causing the reaction ^{9, 10}. AR also shows the global impact on a person's health and economic condition ¹¹. Evidence shows that changes in climate, environmental exposure, family history, and lifestyle factors increase the chances of getting an allergic disorder ^{12, 13}.

A strong association exists between asthma and allergic rhinitis, where many individuals diagnosed with allergic rhinitis also suffer from asthma, and vice versa ¹⁴. Research estimates suggest that nearly 80% of individuals with asthma also experience allergic rhinitis. Research data shows allergic rhinitis increased theatrically daily in Europe (Danish), from 19 to 32% and for Americans 10 to 30%, respectively ^{15, 16}.

Avoiding is the best treatment for AR. If medication is the preferred course of action, managing allergic rhinitis often involves utilizing antihistamines, nasal corticosteroids, decongestants, and immunotherapy as commonly employed treatments. Lifestyle changes such as keeping the home clean, avoiding tobacco smoke, and reducing exposure to pets can also help manage allergic rhinitis ^{17, 18}. H1 antihistamines are extensively used in reliving seasonal and perennial AR, urticaria, mild-moderate seasonal asthma, angioedema, etc. ^{19, 20}. Second-generation antihistamines are characterized by their high molecular weight, low lipid solubility, and reduced affinity for cerebral H1 receptors ^{21, 22}. It is also analyzed that whenever an accident happens, either road or air, excluding alcohol/drug, there is always first-generation H1-antihistaminic found in post-partum report ^{23, 24}.

Levocetirizine: Levocetirizine hydrochloride (LCH) is a second-generation antihistamine used to cure AR, urticaria, and other allergic conditions. It comes in different dosage forms, such as tablets, oral solutions, and nasal sprays. In May 2007, the United States Food and Drug Administration (FDA) approved Levocetirizine, followed by a successful New Drug Application (NDA) submission ²⁵. Dr. Reddy’s Laboratory has launched Levocetirizine tablets (5mg) and different doses. By blocking histamine, LCH can relieve AR indications and progress quality of life for people with allergies ²⁶. Levocetirizine, marketed as Xyzal®, is a second-generation antihistamine that Levocetirizine is the active R-enantiomer of cetirizine. Its chemical formula is C21H25ClN2O3, and its chemical name is 2-(2-{4-[(R)-(4-chlorophenyl) (phenyl) methyl] piperazin-1-yl} ethoxy) acetic Acid. It belongs to the piperazine class of medications ²⁷ as shown below in Fig. 1.

FIG. 1: CHEMICAL STRUCTURE OF LEVOCETIRIZINE

It is selective, potent, non-sedating, and acts as an inverse agonist Levocetirizine is a more potent and selective antagonist of the histamine H1 receptor ^{28, 29, 30, 31}. Levocetirizine is prescribed to manage various allergic conditions such as allergic rhinitis, atopic dermatitis, acute and chronic idiopathic urticaria, and insect bites and stings ^{32, 33}. Additionally, to treat hay fever, seasonal asthma accompanied by allergic rhinitis, and some respiratory infections (although no scientific evidence supports its use for the latter) ^{34, 35}. Also, have a large dose tolerance for antihistamine, antiangiogenic, and additional anti-inflammatory activities with no side effects on the cardiac system ^{36, 37}. Nasal spray of LCH was also developed. Rathanan et al. ³⁸.

Nasal In-situ Gel: Nasal in-situ gel is a unique drug delivery system designed to undergo gelation in-situ, i.e., inside the nasal cavity, after administration in the liquid form. This type of preparation with polymer can form gel when triggered by a specific condition like body temperature, which enables the drug to remain in the nasal cavity for longer, providing sustained drug release. Additionally, the gel has mucoadhesive properties, which allow it to adhere to the nasal mucosa and facilitates the drug's absorption ³⁹.

The in-situ gel designed for nasal administration presents a promising avenue for managing diverse respiratory conditions such as AR, nasal congestion, and other related ailments. The advantages of this system include improved drug delivery, reduced dosing frequency, increased patient compliance, and reduced side effects. This formulation also increased permeation ⁴⁰.

Different stimuli, such as changes in temperature, pH, or ions 41 can trigger gelation of in-situ gelling systems (ISGs). The nasal in-situ gel system enables a prolonged and sustained drug release, improving the holding period and bioavailability ⁴². Carbopol is a commonly used polymer for in-situ nasal gel because it converts solution into viscid form at a low concentration when the pH is neutralized. It is a water-soluble polymer that provides mucoadhesive properties ⁴³.

Hypromellose (HPMC) is another polymer commonly used in intra-nasal preparation, in addition to Carbopol. HPMC is a water-soluble polymer that forms a gel-like substance upon contact with aqueous fluids. Utilizing these polymers as viscosity boosters or gelling agents can broaden the rheological attributes of the gel, consequently leading to sustained drug delivery ⁴⁴.

Artificial nasal fluid (ANF) is a solution designed to mimic fluid composition in the nasal chamber. The composition of ANF may vary depending on the specific application, but a commonly used composition is as follows:

• 150 ± 32 mM sodium (Na+)
• 41 ± 18 mM potassium (K+)
• 4 ± 2 mM calcium (Ca2+)
• pH 6

Other ions and compounds such as chloride (Cl-), bicarbonate (HCO3-), magnesium (Mg2+), and glucose may also be included in ANF, depending on the specific application ⁴⁵.

EXPERIMENT:

Material: LCH was procured from GRANULE INDIA LIMITED, Tadi, Visakhapatnam, as a gift sample. Many additives used for the preparation of the in-situ nasal gel include Carbopol 941 (from LobaChemiePvt. Ltd., Mumbai), HPMC K15 (from LobaChemiePvt. Ltd., Mumbai), Benzalkonium chloride (from Ases chemicals works, Jodhpur), glycerine (from Ases chemicals works, Jodhpur), distilled water (prepared in-house), dialysis membrane (from Hi–Media, Mumbai), Whatman’s filter paper-42 (from Whatman Int. Ltd, England), and other analytical/IP/equivalent grade chemicals and solvents. Equipment used includes a digital pH meter (Hanna, PHep), Brook field viscometer (book field LV DV-II+ pro), magnetic stirrer (Remi, 1MLH), and weighing balance (Adair Dutt-FX 200).

Preparation of Levocetirizine Hydrochloride Gel: The study used a full 3² factorial design to check the properties of two influencing variables, Carbopol 941 and HPMC K15, on the gel's properties. The main cause of developing the intra nasal gel preparation was to create a user-friendly aqueous formulation that can be easily self-administered and would undergo instant gelation on pH changes. Introductory trials were conducted to evaluate the gelation properties of Carbopol 941 at concentrations ranging from 0.4% to 0.8% w/v and the viscosity-enhancing properties of HPMC K15 at the same concentration range.

These concentrations were used to prepare nine different formulations (F1 to F9) of the LCH nasal in-situ gel. The levocetirizine drug was dissolved in distilled water to prepare the in-situ gel formulation. Separately, HPMC K15 was triturated with glycerine and mixed with buffer salt solution before hydrating. Carbopol 941 was then added to the solution and allowed to hydrate overnight. The solution was stirred at 500 RPM for 2 hours on a magnetic stirrer, and the drug was added with continuous stirring until thoroughly mixed. Distilled water is introduced to the solution to achieve the final volume, followed by a preservative. The pH of the solution was then adjusted to 4 using either sodium hydroxide or hydrochloric acid. Upon administration, the solution rapidly gelled when it reached the nasal mucosal membrane, attributed to the pH alteration. The composition of various formulations of LCH was revealed in Table 1.

TABLE 1: THE COMPOSITION OF VARIOUS FORMULATIONS OF LEVOCETIRIZINE HYDROCHLORIDE

Evaluation:

Physical Examination ^{46, 47, 48}: The color, homogeneity, and texture of the prepared LCH in-situ gel formulations were visually examined. The observations are presented in Table 2.

Measurement of pH using Digital pH Meter ⁴⁹: The pH of the In-Situ gel formulations was measured using a digital pH meter. To do this, 1gm of gel was dissolved in 100 ml of distilled water. The pH of each formulation was measured in triplicate, and the mean ± standard deviation (SD) was calculated. The observations are revealed in Table 2. 

Rheologic Studies with Brookfield Viscosimeter ^{50, 51}: The rheology of the LCH In-Situ gel was assessed using a Brookfield viscometer with spindle 96 for the gel and spindle 62 for the solution, both before and after gelation).

The readings were noted down and the averages of three readings were calculated. The viscosity values of the LCH In-Situ gel formulations at pH 6.4 and pH 7.4 were recorded and are presented in Table 2.

Gelling Capacity ⁵²: It was determined by adding the formulation to simulated nasal fluid and evaluated. The observations are presented in Table 2.

Drug Content Determination ⁵³: To determine the drug quantity of the LCH In-Situ gel formulations, 1 ml of each formulation was accurately pipetted out and diluted with distilled water to up to 100 ml.

Then, a 5 ml aliquot of the diluted solution was withdrawn and further diluted with 25 ml of distilled water. The absorbance of LCH in each diluted solution was then measured at 231 nm. The drug quantity of each formulation was calculated using a standard calibration curve of LCH, and the results are presented in Table 2.

Measurement of Gel Strength ⁵⁴: To determine the gel strength of the nasal in-situ gel formulations, A volume of 100 ml was measured using a graduated cylinder, and the sample 35 g used was positioned on top of the gelled formulation. The time noted in seconds to penetrate 5 cm into the cylinder was recorded as the gel strength. The gel strength values for all the formulations are presented in Table 2.

Determination of Mucoadhesive Strength ⁵⁵: Using tape, two glass slides were fixed with a piece of egg membrane. 50mg of gel was placed on one slide, then under the height-adjustable pan. The second slide was fixed inverted position and held against the first slide for 2 minutes to ensure intimate contact. The weight was essential for separation was then measured. The mucoadhesive strength was determined, as mentioned in Table 2.

Mucoadhesive strength (dynes/cm²) = mg/A

Where: m= gm wt. for detachment, g = acceleration 980cm/s²,A = Exposed are of membrane

Drug Release ^{56, 57, 58}: The in-vitro issue of medicine from the in-situ gel formulations was evaluated using two-chamber diffusion cells with a dialysis membrane of molecular weight cut off 1200-1400 KDa. The diffusion cell had a diameter of 1.5 cm and a capacity of 20 ml, with an upper cylindrical compartment open from above and a diffusion membrane at its base. Two milliliters of in-situ gel loaded with the drug were placed in the donor section, and 20 ml of phosphate buffer pH 6.4 was located in the receptor section.

The temperature was maintained at 37°C ± 0.5°C during the experiment, and the content was stirred at 50 rpm using a magnetic stirrer. Subsequently, samples of 1 ml were extracted from the receiver chamber initially after 30 minutes and subsequently at hourly intervals for up to 6 hours, followed by replacement with a fresh medium of equal volume.

These extracted samples were appropriately diluted and subjected to UV spectrophotometric analysis at 231 nm for drug quantification. The in-vitro release was conducted for a total duration of 6 hours. The drug release profiles obtained from the in-situ gel formulations were compared with the theoretical release profile, as presented in Table 3.

RESULTS AND DISCUSSION: The physical characteristics of the prepared Levocetirizine Hydrochloride gelling systems, such as texture, homogeneity, and color, were found satisfactory for all formulations, as shown in Table 2. The pH of the nasal mucosal membrane in a normal physiological state ranges from 5.5 to 6.5. However, in the case of A R, the pH of the nasal mucosal membrane increases to 7.2-8.3 ⁵⁹. The physical characteristics of the prepared gelling system of Levocetirizine Hydrochloride were found to be good, including texture, homogeneity, and colorlessness. The pH of all gels was within the acceptable range of 6.2±0.10 to 6.4±0.44 for nasal mucosa tolerance. Due to hydrogen bonding, lower pH values (<7) provide better mucoadhesive properties and longer retention in the nasal chamber.

The drug amount of the various formulations were within the range of 87.1 ± 0.049% to 109.76 ± 0.041%. The gel strength was found between 14 ± 0.35 secs to 71 ± 0.55secs, while the Mucoadhesive Strength was observed between 854 ± 0.33 dynes/cm2 to 5517 ± 1.88 dynes/cm².

The viscosity values were found between 200-1400 cps at pH 6.4 and 2000-5500 cps at pH 7.4, at 20 rpm. The average effect of the concentration of the suspending or thickening agent (HPMC K15M) and the gelling agent (Carbopol 941) of the prepared LCH nasal in-situ gel at 20 rpm at pH 6.4 and pH 7.4 are shown in Fig 2.

The analysis of viscosity data through ANOVA revealed that an increase in the concentration of HPMC K15M from 0.4% to 0.8% resulted in an insignificant increase in viscosity at pH 6.4 (P=0.57; df 2,6; F=0.61; Fcrit=5.14).

Likewise, HPMC K15M different concentrations from 0.5% to 1.5% resulted in an insignificant increase in viscosity at pH 7.4 (P=0.55; df 2, 6; F=0.64; Fcrit=5.14).

However, when Carbopol 941, different concentrations from 0.4% to 0.8% resulted in a substantial rise in viscosity at pH 6.4 (P=0.015; df 2, 6; F=8.97; Fcrit=5.14). Similarly, when Carbopol 941concentraion increased from 0.4% to 0.8% resulted in a substantial rise in viscosity at pH 7.4 (P=0.010; df 2, 6; F=10.56; Fcrit=5.14).

Physical Examination, pH, Gel Strength, Gelling Capacity, Rheology and Mucoadhesive Strength:

TABLE 2: PHYSICAL CHARACTERISTICS OF PREPARED GELLING SYSTEM OF LEVOCETIRIZINE HYDROCHLORIDE

*Average of three determinations. Note: Few minutes of application, the +gel dissolves rapidly, while the ++gel shows immediate gelation and remains stable for a few hours. The +++gel shows immediate gelation and remains stable for extended periods.

FIG. 2: AVERAGE CONCENTRATION-DEPENDENT EFFECT OF HPMC K15M AND CARBOPOL 941 ON VISCOSITY AT 20 RPM AT PH 6.4, 7.4 RESPECTIVELY

In-vitro Release Study: The descending order of LCH release from all in-situ gel formulations was as follows: F1 > F2 > F3 > F5 > F6 > F4 > F8 > F7 > F9.

FIG. 3: IN-VITRO RELEASE OF LEVOCETIRIZINE HYDROCHLORIDE FROM DIFFERENT FORMULATIONS

The release of LCH after 6 hours was 91.45%, 88.42%, 85.06%, 84.34%, 81.5%, 80.98%, 80.05%, 79.46%, and 79.07% for formulations F1 to F9, respectively, as illustrated in Fig. 3.

To compare the release contour of Levocetirizine Hydrochloride Nasal in-situ gel with a theoretical profile, the similarity factor ƒ2 was used. Formulations F1 to F9 had similarity factor values of 43.32, 47.37, 51.80, 51.71, 52.44, 55.75, 64.95, 67.91, and 80.23, respectively.

This indicates that except for the first two formulations, the rest had a similar release profile to the desired theoretical profile, with a similarity factor value of more than 50. Average concentration-dependent effect of HPMC K15M and Carbopol 941 on in-vitro release of levocetirizine hydrochloride from different formulations at 360 and 240 min respectively in Fig. 4.

FIG. 4: AVERAGE CONCENTRATION-DEPENDENT EFFECT OF HPMC K15M AND CARBOPOL 941ON IN-VITRO RELEASE OF LEVOCETIRIZINE HYDROCHLORIDE FROM DIFFERENT FORMULATIONS AT 360 AND 240 MIN, RESPECTIVELY

TABLE 3: IN-VITRO DRUG RELEASE OF IN-SITU GEL ALONG WITH THEORETICAL RELEASE PROFILE

The release date at 360 minutes was analyzed using ANOVA, which revealed that an increase in the concentration of HPMC K15M from 0.4% to 0.8% resulted in an insignificant (P=0.80; df 2, 6; F=0.22; Fcrit=5.14) decrease in drug liberation. On the other side, when Carbopol 941concentraion increased from 0.4% to 0.8% resulted in a significant (P=0.006; df 2, 6; F=13.23; Fcrit=5.14) decrease in drug liberation. The release date at 240 minutes was analyzed using ANOVA, revealing that an increase in the concentration of HPMC K15M from 0.4% to 0.8% resulted in an insignificant (P=0.81; df 2,6; F=0.20; Fcrit=5.14) decrease in drug release. On the other hand, when Carbopol 941 concentration increased 941 from 0.4% to 0.8% resulted in a significant (P=0.005; df 2,6; F=14.00; Fcrit=5.14) decrease in drug release.

Kinetic Treatment of Release Data: Table 4 shows the release kinetics of different formulations obtained by fitting the raw dissolution data of drug passage in solution into various release models, including zero-order, first-order, Higuchi's, Hixson Crowell's, and Korsmeyer's Peppas equations.

TABLE 4: THE RELEASE KINETICS OF VARIOUS FORMULATIONS

Formulation F5, F7, F8 and F9 the suitable model was Zero Order model with correlation coefficient of 0.9902, 0.9956, 0.9971 and 0.9997, respectively.

Data Analysis:

Multiple Regression Analysis: Multiple linear regression analysis (MLRA) was used to generate a second-order statistical model that included interaction and polynomial terms for all the response variables. The following equations were derived to describe the relationship between drug release (Y) and concentration of Carbopol 941 (X₁) and HPMC K15M (X₂):

Model for Drug Release:

At 6 Hours:

Y₃₆₀ = 115.63-93.84X₁+12.81X₂+41.17X₁²-33.76X₂²+37.46X₁X₂

(R² = 0.944)

Model for Drug Release:

At 4 Hours:

Y₂₄₀ = 59.79+53.55X₁+8.97X₂-68.62X₁²-10.97X₂²-9.46X₁X₂

(R² = 0.890)

The model exhibited a good fit for the responses as specified by the high R2 values. The amount of Carbopol 941 (X1) and HPMC K15M (X2) significantly influenced drug release.

Response Surface Methodology ⁶⁰: The 3-dimensional response surface for percentage drug release at 6 hours and 4 hours is shown in Fig. 5 and the 2D contour plot is in Fig. 6.

The 3-dimensional response surface for the viscosity of the gel at pH 7.4 and pH 6.4 is shown in Fig. 7. The contour plot showing the impact of the varying concentration of Carbopol 941 and HPMC K15M on the viscosity of Levocetirizine Hydrochloride in-situ nasal gel at pH 7.4 (a) and pH 6.4 (b) at 20 RPM is revealed in Fig. 8.

FIG. 5: THREE-DIMENSIONAL RESPONSE SURFACE PLOTS PRESENTING THE CONCENTRATION-DEPENDENT EFFECT OF CARBOPOL 941 AND HPMC K15M ON THE PERCENT DRUG RELEASE OF LCD

FIG. 6: CONTOUR PLOT PRESENTING CONCENTRATION-DEPENDENT EFFECT OF CARBOPOL 941 AND HPMC K15M ON RELEASE OF LCH FROM IN-SITU NASAL GEL AT 6 HOURS (A) AND AT 4 HOURS (B)

The following equations were derived to describe the relationship between Gel Viscosity (Y) and concentration of Carbopol 941 (X₁) and HPMC K15M (X₂):

Model for Gel Viscosity:

At 7.4 pH 20 RPM:

Y_7.4 = 2114.66-4197.50X₁-588.33X₂+5112.50X₁²-325X₂²+6125X₁X₂

(R² = 0.981)

Model for Gel Viscosity:

At 6.4 pH 20 RPM:

Y_6.4 = 375.58+501.25X₁-1893.75X₂-700X₁²+525X₂²+3306.25X₁X₂

(R² = 0.999)

FIG. 7: THREE-DIMENSIONAL RESPONSE SURFACE PLOTS SHOWING CONCENTRATION-DEPENDENT EFFECT OF CARBOPOL 941 AND HPMC K15M ON VISCOSITY OF LEVOCETIRIZINE HYDROCHLORIDE IN-SITU NASAL GEL AT 7.4 PH (A) AND AT 7.4 PH (B) 20 RPM

FIG. 8: CONCENTRATION-RESPONSE CONTOUR PLOT OF CARBOPOL 941 AND HPMC K15M ON VISCOSITY OF LEVOCETIRIZINE HYDROCHLORIDE IN-SITU NASAL GEL AT 7.4 PH (A) AND AT 7.4 PH (B) 20 RPM

Design Space ⁶¹:

Building of Design Space: The proposed design space comprises the overlap region of ranges for percentage drug release (6 hours and 4 hours) for concentration is 0.765-0.800 for both Carbopol 941 and HPMC K15M.

Check Point Formulation: Validation of Model for Design Space: A formulation was prepared with X₁: amount of Carbopol 941 i.e. 0.775 and X₂: amount of HPMC K15M i.e. 0.775. The in-situ nasal gel was then subjected to release studies to assess drug release at 6 hours and 4 hours. The actual and predicted drug release values were compared by determining % bias or residual. The formula for the calculation of % bias/residual is as follows:

% Bias/Residual = (Predicted value – Actual value)/Predicted value × 100

The predicted response using the regression equation, the actual response and residuals are presented in Table 5.

TABLE 5: RESULTS OF EVALUATION OF THE CHECKPOINT FORMULATION

The close resemblance between the actual and the predicted response indicates the validity of the generated models.

Stability Studies of the Optimized Batch as per ICH Guidelines: Q1 “Stability Testing of New Drug Substances and Product” ⁶²:

Accelerated Stability Testing: The optimized checkpoint formulation was filled in plastic dropping bottle container and stored at 40^oC temperature and 75% R.H. for 4 weeks. Samples were withdrawn at 7-day time intervals and evaluated. In-vitro release of the intra nasal gel preparation of LCH from optimized check point formulation stored under accelerated stability conditions shown in Fig. 9.

FIG. 9: IN-VITRO RELEASE OF THE NASAL GEL OF LCH FROM OPTIMIZED CHECKPOINT FORMULATION STORED UNDER ACCELERATED STABILITY CONDITION

Approximation of Shelf Life: Using the Bracket method, on subjecting the value into the equation:

t_s = exp [20/0.00199(1/298-1/313)] x 120

= days as 527.21 days or 17.57 months or 1.44 yrs.

RESULTS AND DISCUSSION: The optimized formulation demonstrated good physical stability as no discoloration or physical change occurred after storage. All parameters met the acceptance criteria.

Gel Sterility Test ⁶³: Fluid thioglycollate medium (FTG) for anaerobic/aerobic bacteria incubated at 30º to 35º while Soybean-casein digest medium (SBCD) for the fungi and aerobic bacteria at 20º to, 25º recpectively.

Outcomes: Three different culture media were prepared as test, control, and negative. In the test (FTG and SBCD), 1 ml of the gel was mixed aseptically and incubated for 7 days. Similarly, in the control (FTG and SBCD) medium, normal tap water introduce to cross-check the observation. A negative was also prepared in which nothing was to be added.

After 7 days, there is growth in “control” as the FTG became more intense red and SBCD precipitated, while no growth was observed in the test and negative.

Test for Isotonicity ⁶⁴: The isotonicity of nasal fluid is equivalent to that of a 0.9% sodium chloride solution, which is the same as the isotonicity value of blood.

The optimized formulation is mixed with little blood droplets. It was observed microscopically at a magnification of 45x, and the shape of the blood cells was compared. No ruptures occurred in the red blood cells.

CONCLUSION: A complete 32-factorial design was used to examine the impact of two self-determining variables, namely the amount of Carbopol 941 and HPMC K14M, on the percentage of the drug issue, viscosity, and mucoadhesive strength. After that, optimization was carried out.

To address the challenges associated with other drug dosage forms, such as limited availability of medicines in the nasal mucosal area, adding water-soluble polymers to increase the viscidness of the dosage form is one solution. Another approach involves increasing drug residence time by transforming polymeric solutions into in-situ gel due to pH and physiological factors. Levocetirizine has been confirmed to have a stronger and more long-lasting effect than other antihistamines like loratadine or cetirizine.

Additionally, Levocetirizine has a lower risk of causing drowsiness and other side effects, which can be a significant concern for people who need to remain alert and focused during the day also has a quick response, which means that it starts working more quickly to relieve allergy symptoms. The use of pH triggered in-situ gelling system for LCH provides several advantages, such as improved residence time, sustained drug release, and targeted drug discharge to the nasal mucosal membrane. The incorporation of Carbopol 941 and HPMC K14M in the formulation facilitates gel formation upon coming in contact with the physiological pH of the nasal mucosal membrane, which aids in extending drug availability and reducing dosing frequency. The study underscores the significance of fine-tuning the polymer concentrations employed in the formulation for achieving the desired viscosity and drug liberation profile. In conclusion, the in-situ gel system developed in this study exhibits the potential for enhancing therapeutic outcomes and promoting patient adherence in managing AR.

ACKNOWLEDGEMENT: The authors thank the associate professor, B N College, for encouraging and supporting the manuscript’s preparation.

CONFLICTS OF INTEREST: The Authors declared no conflict of interest.

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    How to cite this article:

    Khatri U, Bharkatiya M and Saini S: Formulation and evaluation of pH trigger nasal in-situ gel of levocetirizine hydrochloride for localized treatment of allergic rhinitis. Int J Pharm Sci & Res 2023; 14(5): 2573-85. doi: 10.13040/IJPSR.0975-8232.14(5).2573-85.

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