INTRODUCTION:

Simethiconeα-(trimethylsilyl) ω methylpoly [oxy (dimethylsilylene)], is present with silicon dioxide to enhance defoaming properties of silicon. It is used as an anti-foaming agent to minimize bloating, distress, or pain due to excessive gas. ¹It is also spelled as Rugby. It is available in the Indian market as tablet dosage form in the brand names of Wind-eze, WindSetlers, Infacol. ²

Simethicone Cinitapride

Mode of action of Simethicone also reduces gas bubbles that obstruct visualization in various gastrointestinal procedures such as radiography, gastroscopy, and colonoscopy.³ Cintapride Cinitapride, chemically 4-amino-N-[3-(Cyclohexan-1-yl-methyl)-4-piperidinyl]-2-ethoxy-5-nitrobenzamideure 1), is a substituted benzamide gastroenteric prokinetic agent acting via complex, but synergistic effects on serotonergic 5-HT2 and 5-HT4 receptor and dopaminergic D2 receptors in the neuronal synapses of the myenteric plexi.⁴ It is also spelled as Citipride. It is available in the Indian market as tablet dosage form in the brand names of Kinpride, Cintrapro.⁵ Mode of action of Cinitapride works by increasing the concentration of a chemical known as acetylcholine that helps in increasing the contractions of the stomach muscles. This helps in faster emptying of the food contents from the stomach to the intestine. It also eases bowel movements by increasing the coordination between the stomach and intestine⁶. In this research work Simethicone and Cinitapride in combination is selected as model of drug. In literature several analytical methods have been reported for the estimation of Simethicone and Cinitapride, either in single or in combined dosage form with other drugs. The combination of these two drugs has not been mentioned in literature neither the UV- Spectroscopy or RP-HPLC method. So in the present work an attempt has been made to develop simple easy to perform, accurate, cost effective and rapid UV-Spectroscopy and RP-HPLC methods for the estimation of CNP and SMC in bulk and combined oral dosage form and also to validate the developed methods.

MATERIALS AND METHODS:

Materials and Reagents:

A gift sample of Simethicone (Gift sample obtained from Rugby laboratories 17177 N laurel park drive, suite 233 livonia, MI 48152) Ether. Cinitapride (Gift sample obtained from (Aar Ess Remedies Private Limited, C-28, Sector-65, Noida-201301, Uttar Pradesh, India) Dimethyl sulfoxide. All the above reagents were of analytical grade and were procured Loba Chem., SD Fine Chemicals and other manufacturers from local market.

Apparatus and Equipment Required:

UV-Vis double beam spectrophotometer (Model; Shimadzu: 1700S, Japan), Electric Sonicator, Volumetric flasks (10ml, 50ml, 100ml), Calibrated analytical pipettes, Electronic digital balance (Techno, Mumbai) RP-HPLC Method Electric Sonicator Volumetric flasks (10ml, 50ml,100ml), Calibrated analytical pipettes. Electronic digital balance (Techno, Mumbai)

Preparation of concentration range (Beer’s limit): Determination of concentration range which obeys the Lambert and Beer’s law is necessary for accuracy and reproducibility in Spectrophotometric analysis for quantitative determination of any drug. For this; Simethicone stock solution (100µg/ml) was prepared using pure drug in Ether.

Further dilutions were made using. 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, 1.2ml, 1.4ml, 1.6ml and 1.8ml of above solution was transferred to a series of 10ml volumetric flasks this gave a series of concentrations ranging from 2 to18 µg/ml of Ether. The final volume was made up to 10ml mark using Ether, sonicated for 5 minutes. The resultant solutions were measured using a Spectroscopy at wavelength of 276.8nm against a reagent blank. A calibration curve was plotted with concentration against absorbance. From the graph it was clear that Beer’s law was obeyed in concentration range of 2-14µg/ml and deviation was observed above these concentrations.

Determination Concentration Range (Beer’s limit): Determination of concentration range which obeys the Lambert and Beer’s law is necessary for accuracy and reproducibility in Spectrophotometric analysis for quantitative determination of any drug. For this; Cinitapride stock solution (100µg/ml) was prepared using pure drug in Dimethylsulfoxide.

Further dilutions were made using 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, 1.2ml, 1.4ml, 1.6ml and 1.8ml of above solution was transferred to a series of 10ml volumetric flasks this gave a series of concentrations ranging from 2to18 µg/ml of Cinitapride. The final volume was made up to 10ml mark using Dimethylsulfoxide, sonicated for 5 minutes. The resultant solutions were measured using a RP-HPLC at wavelength of 260.2nm against a reagent blank. A calibration curve was plotted with concentration against absorbance. From the graph it was clear that Beer’s law was obeyed in concentration range of 2-14µg/ml and deviation was observed above these concentrations.

Preparation of Standard calibration curve: 100 mg of pure Simethicone drug was dissolved in little quantity of Ether in a 100ml volumetric flask, the volume was made up to the mark using Ether. The solution was sonicated for 10minutes. This gave a Simethicone solution with concentration of 1mg/ml (1000µg/ml). 10ml of this solution was further diluted 100ml in volumetric flask using to obtain a concentration of 100µg/ml. Further dilutions were made using 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6 and 1.8ml solution was transferred to a series of 10ml volumetric flasks (to Obtain a series of concentrations ranging from 2-14 µg/ml of). The final volume was made up to10ml mark using of Ether, sonicated for 5 minutes, the solutions were measured an of 276.8nm against a reagent blank. A calibration curve was plotted with concentration against absorbance

Preparation of Standard Calibration Curve: 100 mg of pure Cinitapride drug was dissolved in little quantity of Dimethylsulfoxide in a 100ml volumetric flask, the volume was made up to the mark using Dimethylsulfoxide. The solution was sonicated for 10 minutes. This gave a Cinitapride solution with concentration of 1mg/ml (1000µg/ml). 10ml of this solution was further diluted 100ml in volumetric flask using to obtain a concentration of 100µg/ml. Further dilutions were made using 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6 and 1.8ml solution was transferred to a series of 10ml volumetric flasks (to Obtain a series of concentrations ranging from 2-14 µg/ml of). The final volume was made up to10ml mark using of Dimethylsulfoxide, sonicated for 5 minutes, the solutions were measured a of 260.2nm against a reagent blank. A calibration curve was plotted with concentration against absorbance

Development of Simethicone in Tablet Dosage Forms: Twenty tablets were weighed accurately and powdered. The Tablet powder equivalent to 100 mg of Simethicone was transferred into a 100mL volumetric flask and dissolved in little quantity of Ether. Then the solution was sonicated for 30 minutes and filtered using Whatman filter paper No#41. The filtrate so obtained was diluted with Ether produce 100ml. Further dilutions were made with Ether to get required concentrations within Beer’s - Lambert limits. The resultant solutions were measured at wavelength of 276.8nm against are reagent blank. The concentration of drug was calculated with the help of standard calibration curve

Development of Cinitapride in tablet dosage forms: Twenty tablets were weighed accurately and powdered. The Tablet powder equivalent to 100 mg of Cinitapride was transferred into a 100mL volumetric flask and dissolved in little quantity of Dimethylsulfoxide. Then the solution was sonicated for 30minutes and filtered using Whatman filter paper No#41. The filtrate so obtained was diluted with Dimethylsulfoxide produce 100ml. Further dilutions were made with Dimethylsulfoxide to get required concentrations within Beer’s - Lambert limits. The resultant solutions were measured at wavelength of 260.2nm against are reagent blank. The concentration of drug was calculated with the help of standard calibration curve.

Analytical Method Validation:

Validation of an analytical procedure is the process by which it is established, by laboratory studies, that the performance characteristics of the procedure meet the requirements for its intended use. All analytical methods that are intended to be used for analyzing any clinical samples will need to be validated. Validation of analytical methods is an essential but time consuming activity for most analytical development laboratories. It is therefore important to wave length. Understand the requirements of method validation in more detail and the options that are available to allow for optimal utilization of analytical resources in a development laboratory.

RESULTS AND DISCUSSION:

Results of Simethicone:

Results of Determination of Beer's Limit:

Table:1 showing Beer’s range for Simethicone:

S. NO.	Concentration in µg/ml	**Absorbance at λmax 276.8nm
1.	0.0	0.000
2.	2	0.085
3.	4	0.180
4.	6	0.270
5.	8	0.375
6.	10	0.470
7.	12	0.550
8.	14	0.640
9.	16	0.820
10.	18	1.000

(** Average of three determinations).

Figure 1. Showing Beer's limit for Simethicone at 276.8nm

Standard Calibration Curve of Simethicone:

Table 2. Showing Absorbance of Simethicone at Various Concentrations.

Sl.No.	Concentration in µg/ml	Absorbance at λmax 276.8nm
1.	0.0	0.000
2.	2	0.085
3.	4	0.180
4.	6	0.270
5.	8	0.375
6.	10	0.470
7.	12	0.550
8.	14	0.640

Figure 2. Showing standard calibration curve for Simethicone

Results of Determination of Absorption Maxima:

Figure 3. Showing absorption maxima of Simethicone at 276.8 nm

(**Average of six determinations, n=6)

DETERMINATION OF PRECISION:

Table 5. Precision results for Simethicone:

Sl.	Conc. in(µg/ml)	Inter-day Absorbance Mean±S.D**	% C.V	Intra-day Absorbance Mean±S.D**	% C.V
1.	8	0.417±0.009	0.42	0.416±0.021	0.73
2.	10	0.516±0.030	0.41	0.516±0.025	0.32
3.	12	0.609±0.023	0.67	0.612±0.031	0.35

(**Average of three determinations, n=3)

DETERMINATION OF RUGGEDNESS PARAMETERS:

Table 6. Showing Ruggedness parameters:

Parameters	Laboratory	Name of the Instrument	Manufacturer of the chemicals used
Lab.1 With analyst (I)	M.M.U College of Pharmacy, Ramanagara	Shimadzu- (model: 1700S,Japan) double beam UV-Vis Spectrophotometer	S.D Fine chemicals, Mumbai
Lab.2 With analyst (II)	Dr. H.LT. College of Pharmacy, Kengal Channapatna	Systronic UV- Vis double beam Spectrophotometer	Loba chemicals, Mumbai

Table 7. Showing Ruggedness results for Simethicone:

Sl.No.	Brand	Label Claim (mg)	*Lab.1 with analyst I**		*Lab. 2 with analyst II**
			Amount found (mg)	% Recovery ±S.D.**	Amount found (mg)	% Recovery ±S.D.**
1	RUGBY	10	10.9	100.9±0.749	9.97	99.7±0.390

(Lab 1* MMU College of Pharmacy, Lab 2* Dr. HLT College of Pharmacy, **Average of six determinations, n=6)

DETERMINATION OF ROBUSTNESS:

Table 8. Showing Robustness results for Simethicone:

Type	Sl.No.	Conc.In (µg/ml)	Change in temperature		Change in P^H
			+5⁰C	-5⁰C
			Absorbance at 276 8nm Mean± S.D**
Pure	1	8	0.411±0.032	0.412±0.022	0.410±0.017	0.412±0.020
Drug	2	10	0.521±0.028	0.519±0.023	0.520±0.016	0.521±0.042
	3	12	0.615±0.028	0.616±0.032	0.615±0.055	0.614±0.066

(**Average of three determinations, n=3)

Table 9. Showing calibration data for Simethicone at 276.8nm:

Parameters	Calibration data at 276.8nm
λmax	276.8nm
Beer’s law limit (µg/ ml)	2 -14µg/ml
Molar Absorptivity	1.1161Lmol^-1cm^-1
Regression Equation(Y=a+bc)	Y= 0.055X+0.016
Slope (b)	0.01426 to 0.01472
Intercept(a)	- 0.004520 to 0.0016757
Correlation Coefficient (R²)	0.998
Limit of detection (LOD)	1.229µg/ml
Limit of quantitation (LOQ)	5.230µg/ml

RESULTS OF CINITAPRIDE:

Results of Determination of Beer's Limit:

Table 10. showing Beer’s range for Cinitapride:

Sl.No.	Concentration in µg/ml	**Absorbance at λmax 260.2nm
1	0	0
2	2	0.03
3	4	0.061
4	6	0.088
5	8	0.117
6	10	0.146
7	12	0.172
8	14	0.188
9	16	0.21
10	18	0.232

(** Average of three determinations)

Standard Calibration Curve of Cinitapride:

Table 11. Showing absorbance of Cinitapride at various concentrations:

Sl.No.	Concentration in µg/ml	**Absorbance at λmax 260.2nm
1	0	0
2	2	0.030
3	4	0.061
4	6	0.088
5	8	0.117
6	10	0.146
7	12	0.172

ESTIMATION OF CINITAPRIDE IN TABLET DOSAGE FORM:

DETERMINATION OF ACCURACY:

Table 12 Showing Absorption of drug from Tablet Dosage Form:

Volume of Stocks Solution Used	Amount of drug (label claim) (µg/ml)	Absorbance at 260.2nm	Amount of drug found (µg/ml)	Percentage purity found±S.D (%w/w)**
0.2ml	2	0.030	2.011	100.55±0.14
0.6ml	6	0.088	5.635	93.91±1.44
1.0ml	10	0.146	9.977	99.77±0.64
1.4ml	14	0.250	13.176	94.11±0.49
1.8ml	18	0.290	17.144	95.22±1.89

(** Average of three determinations

Table 13. Accuracy results for Cinitapride:

Brands	Initial amount (µg/ml)	Amount of pure drug added (µg/ml)	Amount recovered (µg/ml)	% Recovery ±S.D**
CITAPRIDE	10	8 (80%)	8.022	100.27 ± 0.140
	10	10 (100%)	9.892	98.92 ± 0.222
	10	12 (120%)	12.021	100.17± 0.412

(**Average of six determinations, n=6)

DETERMINATION OF PRECISION:

Table14. Precision results for Cinitapride:

Sl.	Conc. in (µg/ml)	Inter-day absorbance Mean ±S.D **	% C.V	Inter-day absorbance Mean ±S.D **	% C.V
1	8	0.408 ±0.033	0.33	0.407±0.012	0.60
2	10	0.507±0.032	0.32	0.507±0.016	0.21
3	12	0.604±0.58	0.58	0.603±0.022	0.22

(**Average of three determinations, n=3)

DETERMINATION OF RUGGEDNESS PARAMETERS:

Table 15. Showing Ruggedness parameters:

Parameters	Laboratory	Name of the Instrument	Manufacturer of the chemicals used
Lab.1 With analyst (I)	M.M.U College of Pharmacy, Ramanagara	Shimadzu- (model: 1700S, Japan) double beam UV-Vis Spectrophotometer	S.D Fine Chemicals, Mumbai
Lab.2 With analyst (II)	Dr. H.LT. College of Pharmacy, Kengal Channapatna	Systronic UV- Vis double beam Spectrophotometer	Loba Chemicals, Mumbai

Table 16. Showing Ruggedness results for Cinitapride:

Sl.No.	Brand	Label Claim (mg)	*Lab. 1 With analyst I**		*Lab. 1 With analyst II**
			Amount found (mg)	% Recovery ±S.D.**	Amount found (mg)	% Recovery ±S.D**
1	CITAPRIDE	10	10.05	100.5±0.746	9.93	99.3±0.348

(Lab 1* MMU College of Pharmacy, Lab 2* Dr. HLT College of Pharmacy, **Average of six determinations, n=6)

DETERMINATION OF ROBUSTNESS:

Table 17. Showing Robustness results for Cinitapride:

Type	Sl. No.	Conc.in (µd/ml)	Change in temperature		Change in P^H
Type	Sl. No.	Conc.in (µd/ml)	+⁰5 C	-⁰5 C	2 drops of 0.1N NaOH	2drops of 0.1N HCl
			Absorbance at 226.2nm Mean±S.D**
	1	8	0.404±0.025	0.404±0.025	0.402±0.009	0.400±0.018
	2	10	0.514±0.021	0.512±0.016	0.511±0.008	0.512±0.025
	2	12	0.608±0.021	0.609±0.025	0.607±0.047	0.604±0.045

(**Average of three determinations, n=3)

Table 18. showing calibration data for Cinitapride at 260.2nm

Parameters	Calibration Data at 260.2nm
lmax	260.2nm
Beer’s law limit (µg/ml)	2 – 12 (µg/ml)
Molar Absorptivity	1.1154Lmol^-1Cm^-1
Regression Equation (Y=a+bc)	Y = 0.0145×+0.011
Slope (b)	0.01419 to 0.01465
Intercept (a)	-0.004513 to 0.0016750
Correlation Coefficient (R²)	0.9993
Limit of detection (LOD)	1.224 µg/ml
Limit of quantitation (LOQ)	4.088 µg/ml

DISCUSSION:

Determination of Beer’s limit: The Beer’s limit felled in the range of 2-14 µg/ml and 2-12 µg/ml under given experimental conditions.

Determination of absorption maxima: 100 µg/ml stock solution of Simethicone and Cinitapride was prepared and absorbances were measured from 200nm to 340nm. The optimum wave length was found to be 276.8nm and 260.2nm

Assay: Marketed tablets contained Simethicone and Cinitapride were used for the assay. After extraction, proper dilution, measurement, the concentration was determined using standard calibration curve. The amount of drug found in the range of 99.80 – 100.80%w/w and 93.91–99.7% w/w

Method Validation: The proposed method was validated in accordance to ICH guidelines.

a) Accuracy: Percentage of recoveries of Simethicone and Cinitapride in tablets was found in the range of 99.94 –100.30%w/w and 98.92 –100.27%w/w

b) precision: The percent coefficient of variations (% C.V) of Simethicone and Cinitapride was between 0.34-0.60 for intra-day and 0.23-0.62, 0.33-0.58 for intra-day and 0.60-0.22 34-0.60 inter day absorbances.

c) Repeatability: Repeatability was determined by analyzing the sample at the given concentration wavelength for at least six times and it was found that the variability in the results was not more than 0.5%.

d) Reproducibility: The standard solution of Aprocitentan by analyst-I and analyst-II separately. The values obtained were evaluated using F-test and t-test to verify their reproducibility. Calculated value for t-test was found to be less than the tabulated (standard) value it can calculated that no significant difference was observed in the result of analysis.

e) Ruggedness: Ruggedness of the developed method was determined by changing the analytical tools such as laboratory, instruments, analyst and chemicals. The result (in terms of %RSD) of six determinations indicated that there were no significant variations in the data.

f) Robustness: Robustness of the method was established by slightly changing the temperature and PH of the reaction mixture. The data so obtained showed no significant variation in the absorption pattern.

g) Limit of detection and limit of quantitation: were determined from the standard deviation of y – intercepts of six calibration curves and average slope of six calibration curves. LOD and LOQ of Simethicone and Cinitapride was found to be 1.220µg/ml and 5.220µg/ml and 1.224µg/ml and 4.088µg/ml respectively.

CONCLUSION:

A new spectrophotometric method was developed to estimate Semithicone and Cinitapride in pure and tablet dosage forms. Ether and Dimethylsulfoxide solutions of Simethicone and Cinitapride was estimated by using UV Spectroscopy and RP-HPLC (Shimadzu 1700S, Japan) with matched 1cm quartz cell. It showed maximum absorption at the range of 2-14µg/ml 2-12 at 276.8nm and 260.2 with coefficient of correlation (R2) of 0.997. The method so developed was validated according to ICH guide lines for accuracy, precision (inter and intra-day precisions), repeatability, reproducibility, ruggedness, robustness etc. The proposed method was found to be simple, accurate, sensitive, precise, reproducible and rapid. This method can be successfully employed for routine quantitative analysis of Simethicone and Cinitapride in bulk and tablet dosage form.

ACKNOWLEDGMENT:

The authors are thankful to head of the Department of Chemistry, Department of Pharmaceutics, Department of Quality Assurance.

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Received on 28.01.2025 Revised on 11.04.2025

Accepted on 29.05.2025 Published on 19.06.2025

Available online from June 23, 2025

Asian J. Research Chem.2025; 18(3):142-148.

DOI: 10.52711/0974-4150.2025.00023

This work is licensed under a Creative Commons Attribution-Non Commercial-Share Alike 4.0 International License. Creative Commons License.

Volume of stock Solution Used	Amount of drug (label claim) (µg/ml)	Absorbance at 276.8nm	Amount of drug found (µg/ml)	Percentage purity found ±S.D (%w/w)**
0.2ml	2	0.1093	2.019	100.75±0.16
0.6ml	6	0.317	6.043	100.68±1.47
1.0ml	10	0.514	9.985	99.85±0.66
1.4ml	14	0.698	13.984	99.90±0.51

Brands	Initial Amount (µg/ml)	Amount of pure drug added (µg/ml)	Amount recovered (µg/ml)	% Recovery ±S.D**
RUGBY	10	8(80%)	8.031	100.37±0.147
	10	10(100%)	9.901	98.01±0.231
	10	12(120%)	12.035	100.25±0.422