Development And Validation Of Stability Indicating HPLC Method For The Simultaneous Quantification Of Meloxicam And Rizatriptan In Its Pure & Dosage Forms

Pharmaceutical analysis is a critical branch of pharmaceutical sciences that focuses on the identification, quantification, and characterization of drug substances and formulations. It ensures that pharmaceutical products meet the required standards of quality, safety, and efficacy. Analytical methods are essential throughout the drug development process, from raw material testing to finished product evaluation and stability studies. (1–5).Among the various analytical techniques available, High-Performance Liquid Chromatography (HPLC) has emerged as one of the most reliable and widely used methods due to its high sensitivity, specificity, accuracy, and reproducibility. It is particularly useful in the analysis of complex mixtures, impurities, degradation products, and multi-component dosage forms.

MATERIALS AND METHODS:

The experimental work was carried out using various analytical instruments and high quality glassware  to ensure accuracy and precision. A waters 2690 HPLC System separation module and empower 2 software was employed for chromatographic analysis. Additional instruments included a Thermoscientific pH Meter, Dwaraka scientific Thermal Oven, Labman scientific india Ultra-Sonicator, Scaletec Electronic Balance.All volumetric flasks,Pippets and Buretts,Beakers used were of Borosil make.The chemicals used in the study were of analytical grade including Meloxicam and Rizatriptan procured from Qualigens,, Phosphate buffer, Acetic acid, Water, Acetonitrile for HPLC is provided by Qualigens, Methanol for HPLC from Rankem.

METHOD DEVELOPMET

Preparation of Standard Solution.

Accurately weigh and transfer 25mg of Meloxicam, Rizatriptan working standard into a 25ml clean dry volumetric flask add Diluents and sonicate to dissolve it completely and make volume up to the mark with the same solvent. (Stock solution).Further pipette 0.5ml of the above stock solution into a 10ml volumetric flask and dilute up to the mark with Diluents.

Optimization of Column:

SpurcilC18 Column, (250×4.6mm, 5µm) was found to be ideal as it gave good peak shape and resolution at 1.0 ml/min flow.

METHOD VALIDATION

preparation of buffer and mobile phase:

preparation of nah2po4 buffer ph 4.0:

To prepare NAH2PO4 Buffer solution, by adding 6.4Grams of Potassium dihydrogen orthophosphate in 1000ml water. Adjust this solution to pH 4.0 by using sodium hydroxide.

Preparation of mobile phase:

Mix a mixture of above ACN 600ml (60%), 400 ml NAH2PO4 (40%) and degas in ultrasonic water bath for 5 minutes. Filter through 0.45 µ filter under vacuum filtration.

Diluent Preparation:

ACN: NAH2PO4 PH 4 (600:400ml) ratio.

METHOD VALIDATION PARAMETERS:

System Suitability:

Procedure:

Inject 20 µL of the standard, sample into the chromatographic system and measure the areas for the Meloxicam and Rizatriptan peaks and calculate the % Assay by using the formulae.

LINEARITY:

Preparation of stock solution:

Preparation of Level – I-0.2ml Meloxicam and 0.1ml Rizatriptan

Preparation of Level – II-0.4ml Meloxicam and 0.2ml Rizatriptan

Preparation of Level – III-0.6ml Meloxicam and 0.3ml Rizatriptan

Preparation of Level – IV-0.8ml Meloxicam and 0.4ml Rizatriptan

Preparation of Level – V-0.10ml Meloxicam and 0.5ml Rizatriptan

Procedure:

Inject each level into the chromatographic system and measure the peak area.Plot a graph of peak area versus concentration (on X-axis concentration and on Y-axis Peak area) and calculate the correlation coefficient.

PRECISION:

Procedure:

The standard solution was injected for six times and measured the area for all six injections in HPLC. The %RSD for the area of six replicate injections was found to be within the specified limits.

INTERMEDIATE PRECISION/RUGGEDNESS:

Procedure:

The standard solution was injected for six times and measured the area for all six injections in HPLC. The %RSD for the area of six replicate injections was found to be within the specified limits.

ACCURACY:

preparation of 50-150% Sample solution

Accurately weigh and transfer 10 mg of Meloxicam and 5mg Rizatriptan (50%), 20 mg of Meloxicam and 10 mg Rizatriptan (100%), 30 mg of Meloxicam and 15mg Rizatriptane (150%), working standard into a 20ml clean dry volumetric flask add Diluents and sonicate to dissolve it completely and make volume up to the mark with the same solvent. (Stock solution). Further pipette 0.6ml of the above stock solution into a 10ml volumetric flask and dilute up to the mark with Diluents.

Procedure: Inject the standard solution, Accuracy -50%, Accuracy -100% and Accuracy -150% solutions.

Calculate the Amount found and Amount added for Meloxicam and Rizatriptan and calculate the individual recovery and mean recovery values.

RESULTS AND DISCUSSION

Figure:1 Optimized chromatogram(sample)

SYSTEM SUITABILITY:

Figure 2: Chromatogram for system suitability

S.No	Name	RT(min)	Area (µV sec)	Height (µV)	Resolution	USP tailing	USP plate count
1	Rizatriptan	3.099	256214	241189	4.23	1.15	5989
2	Meloxicam	4.205	3654786	3834602		1.32	3998

Table 1: Results of system suitability parameters

VALIDATION PARAMETERS:

ASSAY:

Figure 3: Chromatogram for Standard

Figure 4: Chromatogram for Sample

Drug Name	Label Claim(mg)	% Assay
Rizatriptan	10mg	99.4%
Meloxican	20mg	99.5%

Table 2: Results of Assay for Rizatriptan  and Meloxican

LINEARITY:

S. No	Rizatriptan
	Concentration (µg/ml)	Area
1	10	85438
2	20	170528
3	30	247841
4	40	341085
5	50	427851

 

S. No	Meloxican
	Concentration (µg/ml)	Area
1	20	1282684
2	40	2436854
3	60	3657423
4	80	4873968
5	100	6092543

Table 3: Area of different concentration of Rizatriptan and Meloxican

Figure 5: Calibration graph for Rizatriptan

Figure 6: Calibration graph for Meloxican

Parameters	Rizatriptan	Meloxican
Slope (m)	8524.3	60653
Intercept (c)	983.95	24593
Correlation coefficient (R2)	0.9995	0.9999

Table 4: Analytical performance parameters of Rizatriptan and Meloxican

CONCLUSION

The present study successfully developed and optimized a simple, precise, and reliable RP-HPLC method for the simultaneous estimation of Meloxicam and Rizatriptan in bulk and pharmaceutical dosage forms. The optimized chromatographic conditions, employing a Spurcil C18 column with a mobile phase of acetonitrile and phosphate buffer (60:40, pH 4.0), provided well-resolved peaks with good symmetry and acceptable retention times. System suitability parameters such as theoretical plate count, tailing factor, and resolution were found to be within the prescribed limits, indicating the efficiency of the method. The method validation results demonstrated excellent linearity (R² > 0.999), high precision (%RSD < 2), and satisfactory accuracy with recovery values close to 100%, confirming the reliability of the method for quantitative analysis.

Furthermore, the developed method exhibited good sensitivity with low limits of detection (LOD) and quantification (LOQ), enabling accurate estimation even at trace levels. Robustness studies confirmed that small deliberate variations in chromatographic conditions did not significantly affect the method performance, highlighting its ruggedness for routine use. Forced degradation studies under various stress conditions (acidic, basic, thermal, oxidative, and photolytic) indicated that both drugs undergo degradation to some extent, but the method effectively separated the degradation products from the main peaks, proving its stability-indicating capability. Overall, the developed RP-HPLC method is simple, accurate, precise, robust, and suitable for routine quality control analysis of Meloxicam and Rizatriptan in pharmaceutical formulations.

ACKNOWLEDGEMENT:

The authors express their sincere gratitude to Dr. M. Venkataramana, Principal, Surabhi Dayakar Rao College of Pharmacy, Rimmanaguda, for his constant encouragement, valuable support, and for providing the necessary facilities to carry out this research work successfully.

We extend our heartfelt thanks to Mrs. Soundarya Yerrolla, Assistant Professor, Surabhi Dayakar Rao College of Pharmacy, for her invaluable guidance, continuous motivation, constructive suggestions, and support throughout the completion of this research work.

We are also thankful to all the faculty members, staff, and students of Surabhi Dayakar Rao College of Pharmacy for their cooperation and assistance during the study.

Finally, we express our gratitude to our family members, friends, and everyone who directly or indirectly contributed to the successful completion of this research paper.

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