INTRODUCTION:

In order to ensure and control the quality of bulk medications, pharmaceutical analysis is essential. In analyticachemistry, components in a sample matrix are separated, recognized, and their relative concentrations are calculated¹. A specific area of analytical chemistry is pharmaceutical analysis. The principles of pharmaceutical analysis come from a variety of scientific fields, including physics, microbiology, nuclear science, electronics, etc. The chemical composition of the material is revealed by qualitative analysis.

Establishing the relative abundance species using quantitative analysis or numerical analysis. Prior to conducting a quantitative analysis, qualitative analysis is necessary. Usually, both a qualitative and a quantitative analysis require a separation stage. Two measurements are sufficient to calculate the outcomes of a typical quantitative analysis². The first is the mass or volume of the sample that needs to be examined, and the second is the measurement of a quantity that is proportionate to the amount of analysis that is present in the sample and typically concludes the analysis^3,4.

AIM AND OBJECTIVES:

Generating and validating a UV Spectrophotometric method to estimate a specific medication in pharmaceutical formulations and bulk. The goal of the medication is to create and validate analytical methods that are more specialized, accurate, and precise for a certain analyte.The goal of the current work is to create an accurate and straightforward UV Spectrophotometric method for estimating Domperidone in pharmaceutical formulations^5-7.

Drug Profile:

Chemical description: Category: Antiemetic, Dopaminergic Blocking Agent Molecular formula: C₂₂H₂₄ClN₅O₂Molecular weight: 425.911Melting point: 244.0-246.0 °C

Plan of Work:

· Selecting recently authorized drugs.Acquiring a drug sample.

· performing solubility and analytical research on a chosen drug

· setting up initial spectroscopic parameters

· Optimizing initial spectroscopic parameters, and developing a method for the chosen medication.

· Approval of a newly designed UV Spectrophotometry method in accordance with ICH Guidelines.

· Review of the validation report for an analytical method.

EXPERIMENTAL WORK

Instruments and chemicals use:

UV-Visible Spectrophotometer (LABINDIA), Electronic Balance (Shimadzu-AX-200), Ultra Sonicator(Citizen), Hot air oven – (Model –BIT-29), Bio techniques – India, 0.1M Sodiumhydroxide (NaOH), Methanol, Distilled Water, Acetone and 0.1 N Hydrochloric acid. According to the ICH validation criteria, the analytical method was validated.

Common stock solution creation and producing a working standard solution:

10mg of Domperidone was carefully taken and transferredto a 100ml dry volumetric flask. This volume was then made up with 0.1M NaOH and sonicated for 5 minutes.From the above-mentioned solution, serial dilutions (2ml, 4ml, 6ml, 8ml, 10ml, 12ml, 14ml, 16ml, 18ml and 20ml) were prepared up to 100ml using 0.1M NaOH^9-11.

Calibration curve and Sample setting up:

To obtain 100g/ml of Domperidone, 0.1M NaOH solution was added to the solution after 10mg Domperidone had been weighed, transported, and dissolved in a 100ml flask¹². Aliquots produced from standard solution (0.2, 0.4, 0.6, and 0.8 up to 2ml) were put into a series of 10ml volumetric flasks ,subjected to dilution with 0.1 M NaOH to give a concentration level of 2-20g ml. The aforementioned solutions were scanned against a reagent blank from 200nm to 400nm in wavelength. The absorbance of solution at 233nm in comparison to a blank of 0.1M sodium hydroxide. Plotting absorbance versus concentration allowed for the creation of a calibration curve. Figure 1 displays the Domperidone UV spectrum.

Domperidone Estimation:

For the testing of the medication in bulk, a precisely weighed 10mg test sample was subjected to dissolution in 100ml of 0.1 M NaOH in a volumetric flask. After the proper dilution, the final sample's absorbance value was measured at 233nm against the blank. Twenty Domperidone (10mg) tablets were subjected into a fine powder, and the mixture was properly mixed, in order to analyze the dosage form. 10mg of the drug's powder was moved to a 100ml volumetric flask and dissolved in around 40ml of 0.1M NaoH. Whattman filter paper was used for the filtration process to remove the insoluble ingredients. After the proper dilution, the absorbance value was measured at 233 nm in comparison to blank¹⁴.

Fig.1: Domperidone's UV spectra

RESULTS and discussion:

Linearity: Standard stock solutions ranging from 2 to 20g/ml were utilized to create fresh aliquots, and for this method, 0.1M sodium hydroxide was employed as a blank. At 233nm, the absorbance values of each concentration were measured. The drug displays linearity for this method between 2 and 20g/ml. The findings are presented in Table 1

Table 1: Linearity values ofDomperidone

Values Concentration (µg/ml)	Observed Absr
2	0.083
4	0.179
6	0.249
8	0.339
10	0.412
12	0.500
14	0.580
16	0.643
18	0.728
20	0.820

Precision:

In an intraday trial, the drug's concentration in replicate doses was measured twice on the single day. In an inter-day study, the drug concentration was estimated over the course of two days, expressing the laboratory change over the course of different days. The findings of the intraday and interday precision investigation for the methods%RSD are displayed in Tables 2 and 3.

Table 2: Data on the suggested method's daily repeatability and precision

Concentration (µg/ml)	Absorbance (at 233 nm)
Concentration (µg/ml)	Morning	Evening
8	0.348	0.351
8	0.340	0.389
8	0.349	0.381
8	0.360	0.386
8	0.330	0.343
8	0.360	0.336
Avg	0.3466	0.35926
S. D	0.02479	0.02496
% R.S. D	0.422	0.422

Table 3: Data on the suggested method's intraday precision (reproducibility)

Concentration (µg/ml)	Absorbance range values
Concentration (µg/ml)	Day 1	Day 2
8	0.368	0.321
8	0.380	0.330
8	0.368	0.325
8	0.375	0.325
8	0.386	0.312
8	0.369	0.319
Average	0.3645	0.321333
S.D	0.003036	0.002583
%R.S.D	0.522	0.039

Accuracy:

The accuracy of the established approach, recovery trials were run out in triplicate at three concentration levels of 100%, 125%, and 150%. It was clear from the recovery trials that the procedure is relatively accurate for quantitative determination of tablets because the mathematical findings were in the allowed range¹⁵. (table-4)

Limit of Quantification and Limit of Detection:

Calibration graphs were used to determine the limit of detection and limit of quantification of Domperidone of the suggested approach. L.O.Q and L.O.D were determined as

L.O.D = 3.3 X S.D/S

L.O.Q = 10 X S.D/S

SD - response's least-concentration standard deviation calculated over three replicates and S -slope of the calibration curve.

Table 4: Recovery for the suggested technique

Level Spike range	µg/ml	µg/ml	Percentage recovery	Percentage recovery (Mean)
100 %	8	8.82	95.45	102.8%
100 %	8	9.9	109.72
100%	8	9.5	100.2
125%	10	9.0	96.3	97.83%
125%	10	10.56	100.03
125%	10	9.99	92.9
150%	12	11.81	93.58	96.34%
150%	12	11.91	93.75
150%	12	12.36	99

Table 5: Robustness studies data of proposed method

Concentration ( µg / ml )	Absorbance
Concentration ( µg / ml )	230 nm	233 nm	235 nm
10	0.410	0.426	0.423
10	0.411	0.435	0.433
10	0.413	0.477	0.475
10	0.413	0.455	0.449
10	0.407	0.455	0.447
10	0.413	0.463	0.468
Avg	0.409667	0.466	0.426322
S.D	0.001439	0.008034	0.01190
Percentage of R.S.D	0.0235	0.144	0.185

Ruggedness:

where SD represents of response's least-concentration standard deviation over three replicates and S represents the slope of curve.

Table 6: Ruggedness:

Concentration Values (µg/ml)	Absorbance values (at 233 nm)
	Analyst 1	Analyst 2
16	0.665	0.664
16	0.665	0.668
16	0.660	0.658
16	0.626	0.656
16	0.629	0.682
16	0.670	0.682
Avg	0.6881	0.6886
S.D	0.00590	0.01588
Percentage of R.S.D	0.086	0.259

Table:7 Assay Values (Domperidone Tab)

Type	Value of Claim	Value of Amount Found	% Assay
DOM 10	Domperidone label	Domperidone amount found	98.62

Because the %RSD values for intra-day and inter-day were found to be less than 2%, it was determined that the devised approach was accurate.At each additional concentration, good recoveries (97.11% to 101.31%) of the medication were obtained, proving the method's accuracy. The L.O.D and L.O.Q were discovered as the sub-microgram level, demonstrating the method's sensitivity. Percentages of RSD numbers, which are less than 2%, further demonstrate the robustness and toughness of the technique. As evidenced by the assay's % recovery (98.62%), the amount of medication used was in line with the formulation's label promise^16-19.

SUMMARY:

· UV methods (Four) and RP-HPLC methods (six) have been described for the determination of Domperidone alone and in combination with other medications up to this point, according to a review of the literature. The goal of the current study is to develop and validate a new UV Spectrophotometric method for determining the presence of Domperidone in pharmaceutical formulations and in bulk.

· Domperidone's maximum absorbance (max) in this method is at 233.The medicine complies with Beer's legislation when its concentration is between 2 and 20 g/ml.

· With a correlation value (r2) of 0.998, the drug followed linearity in the concentration range of 2-20 g/ml, indicating strong connection between the response factor and the concentration of standard solutions.

· Because the %RSD values for intra-day and inter-day were observed to be less than 2%, it was determined that the created approach was accurate.

· At each additional concentration, good recoveries (97.11% to 101.31%) of the medication were obtained, proving the method's accuracy.

· The method's sensitivity was determined to be 0.0542916 and 0.045386 g/ml, respectively, for the limits of detection and quantification.

CONCLUSION:

The %RSD figures, which are less than 2%, further prove the method's robustness and toughness. According to % recovery (98.62%), the laboratory findings demonstrate amount of medication found goodin correlation the formulation's label claim.

ACKNOWLEDGEMENTS:

The management and principal of Anurag Pharmacy College, Kodad, and Telangana,India are to be thanked for providing the necessary equipment facilities for research as well as for their encouragement and support.

CONFLICTS OF INTEREST:

The authors declare no conflicts of interest relevant to this article.

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Received on 27.12.2023 Revised on 24.03.2024

Accepted on 20.07.2024 Published on 20.01.2025

Available online from January 27, 2025

Research J. Pharmacy and Technology. 2025;18(1):317-320.

DOI: 10.52711/0974-360X.2025.00049

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