Method Development and Validation for the Analysis of Perindopril Erbumine and Amlodipine Besylate by RP-HPLC in Pure and Pharmaceutical Dosage Form

 

Humera Badar¹, Dr. Ruheena Yasmeen², Fathima Qurratul Ayeen¹, Juveria Badar³

¹Assistant Professor, Department of Pharmaceutical Analysis and Quality Assurance,

Nizam Institute of Pharmacy, Deshmukhi (V), Pochampally (M), Behind Mount Opera,

Yadadri Bhuvanagiri (Dist)-508284, Telangana, India.

²Assistant Professor, Department of Pharm. D (Pharmacy Practice),

Nizam Institute of Pharmacy, Deshmukhi (V), Pochampally (M), Behind Mount Opera,

Yadadri Bhuvanagiri (Dist)-508284, Telangana, India.

³Student, St. Pauls College of Pharmacy, Sy.No.603 and 605, Nagarjuna Sagar Road,

Turkayamjal, Hyderabad, Telangana 501510

 

ABSTRACT:

The aim of the current manuscript is to develop and validate RP-HPLC method for the determination of perindopril erbumine and amlodipine besilate dosage form and to develop and validate calibration curve method and derivative method for the determination of perindopril erbumine and amlodipine besilate in bulk and tablet dosage forms. To develop a simple, rapid and specific RP-HPLC method for the estimation of perindopril erbumine and amlodipine besilate in bulk and combined pharmaceutical dosage forms. To develop a simple, rapid, accurate eco friendly less cost methods for the determination of perindopril erbumine and amlodipine besilate in bulk and tablet dosage forms. To validate the proposed methods in accordance with the analytical parameters mentioned in the ICH guidelines, such as system suitability, accuracy, precision, specificity, linearity, robustness.

 

 

INTRODUCTION:

Perindopril is an ACE inhibitor used in treatment of hypertension andheart failure. It is converted into active metabolite Perindoprilat in the body. It blocks conversion of AT I to AT II and also inhibit degradation bradykinin which is a potent vasodialator, thus decreases peripheral resistance, lowers B.P and produces mild natriuresis. Reduces B.P without increasing heartrate^(1-3). Mostly used in patients having diabetes, naphropathy, CHF left ventricular hypertrophy and post- myocardial infraction. Amlodipine is in a group of drugs called calcium channel blockers.

 

Amlodipine relaxes (widens) blood vessels and improves blood flow. Amlodipine is used to treat high blood pressure (hypertension) or chest pain (angina) and other conditions caused by coronary artery disease^(4-7). In order to develop a simple, reliable and accurate method for the assay of Perindopril erbumine and Amlodipine besilate tablet dosage form by reverse phase HPLC and validate the method for its repeatability and reproducibility. Procurement of raw materials. Establishment of system suitability parameters. Trials for the method development for Perindopril erbumine and Amlodipine besilate. Setting the optimized method. Validation of optimized methods for Perindopril erbumine and Amlodipine besilate^(8-9).

 

INSTRUMENTS:

Preparation of Sample solution:

Weigh 10 tablets and take average weight, then crush the tablets to powder form after take one tablet equivalent weight to weigh accurately about 11.5 mg of sample taken into a 100mL volumetric flask. Add 70mL of diluent, sonicate to dissolve and dilute to volume diluent. Further dilute 1mL to 10mL with the diluent. Further dilute 1mL to 10mL with the diluent. Filter through 0.45μ Nylon syringe filter.

 

MATERIALS AND INSTRUMENTS:

Table 1: List of chemicals used in HPLC Method

S. No	Name	Grade	Manufacturer
1.	Water	HPLC	In house production
2.	Ortho Phospharic Acid	HPLC	Merck
3.	Acetonitrile	HPLC	Merck
4.	Methanol	HPLC	Merck

 

Table 2: List of Apparatus used in HPLC

S. No	Name	Model	Manufacturer
1.	pH meter	-	Eutech
2.	Weighing balance	-	Denver
3.	Ultrasonicator	UCA 701	Unichrome
4.	HPLC	Alliance	Waters 2695- Empower software
5.	Flow rate	1mL/min	--
6.	Pump	Isocratic model	--

 

METHOD VALIDATION:

The validation of HPLC method for the determination of Perindopril Erbumine and Amlodipine Besilate as per the protocol and to demonstrate that the method is appropriate for its intended use was studied for the following parameters. All the validation parameters were carried out according to ICH.

 

System suitability:

The system suitability of developed method was conducted through the validation studies by using 5+6.25µg/mL Perindopril Erbumine and Amlodipine Besilate. System suitability prior to analysis was investigated by checking parameters like tailing factor, retention times and number of theoretical plates. The results were found to be within the limits.

 

RESULTS AND DISCUSSION:

HPLC METHOD:

Determination of Working Wavelength (λmax_):

 

Figure 3: PDA-Spectrum of Perindopril Erbumine and Amlodipine Besilate Optimization of chromatographic conditions Method Development Trails

 

Trail-1

 

Figure 4 : Chromatogram of Trial -1

 

Trial -2

 

Figure 5 : Chromatogram of Trial -2

 

Trial -3

 

Figure 6: Chromatogram of Trial -3

 

Trial -4

 

Figure 7: Chromatogram of Trial -4

Trial 5

 

Figure 8: Chromatogram of Trial -5

 

Trial -6

 

Figure 9: Chromatogram of Trial -6

Trial -7

 

Figure 10: Chromatogram of Trial -7

 

Trail 8: (optimized trail)

 

Figure 11: Chromatogram of Trial -8

 

 

Table 3: Optimized chromatography

 

Table 4: Optimized chromatographic conditions

 

Assay:

Table 5:% Assay of Perndopril Erumine and Amlodipine Besilate

Drug	Avg std Area (n=6)	Avg sample area(n=6)	Avg wt of tab. (mg)	Stdwt (mg)	Sample Wt (mg)	Lable amount (mg)	Std purity	Amount Found (mg)	% assay
Perndopril	1200084	12452132	542.5	100.8	782.5	500	100.1	100.1	100.5
Erumine
Amlodipine	850702	2785329	542.5	100.4	782.5	625	100.1	100.1	100.5
Besilate

 

ANALYTICAL METHOD VALIDATION (HPLC):

The method was validated for its linearity range, accuracy, precision. Method validation is carried out as per ICH guidelines.

 

SUMMARY AND CONCLUSION:

An attempt has been made to develop a new stability indicating validated RP-HPLC method for the estimation of perindopril erbumine and amlodipine besilate in bulk and in dosage form. As the literature survey revealed that only two methods are available for estimation of perindopril erbumine and amlodipine besilate in bulk and in dosage forms so there is a need for a simple, economical and proper method of estimation of perindopril erbumine and amlodipine besilate bulk and in dosage form. Luna C18, 250mm x 4.6mm, 5µm column with UV detector with an injection volume of 10μL was injected and eluted with the mobile phase containing buffer: acetonitrile (50: 50v/v). This is pumped at a flow rate of 1mL/min and detected by UV (210nm) detector. The peaks of perindopril erbumine and amlodipine besilate were eluted at retention times of and 5.042 min respectively. After method was developed, it was validated according to ICH guidelines for system suitability, specificity and linearity, sensitivity parameters, precision, accuracy and robustness studies. the validation results were found well within the limits (% RSD of areas were <2 for assay and recoveries in the range of 98% - 102% for assay, r²>0.999) indicating that the developed method is simple, rapid, accurate, precise, specific, robust and economical and less time consuming.

 

CONCLUSION:

The proposed RP-HPLC, method were suitable methods for the determination of perindopril erbumine and amlodipine besilate in dosage forms. All the parameters of developed methods met the criteria of ICH guidelines for method validation. The developed HPLC method has the following advantages: No tedious extraction procedures were involved. These methods are also having an advantage than reported method of good resolution and with retention time. The developed method has good recovery and sensitivity. The run time required for recording chromatogram was below 10.0mins. Suitable for the analysis of raw materials and formulations. Hence, the developed chromatographic method for perindopril erbumine and amlodipine besilate is said to be rapid, simple, precise, accurate, specific and cost effective that can be effectively applied for the routine analysis.

 

REFERENCES:

1.      Jaiprakash N et.al.,  Development and validation of RP-HPLC method for determination of Perindopril erbumine and Amlodipine besilate in combined tablet dosage form form International Journal of Pharmaceutical Sciences Vol. 4(2) Apr-Jun 2013

2.      Kuldeep Chouhan et al., Validated RP-HPLC Method for simultaneous determination of Perindopril erbumine and Amlodipine besilate and its application in drug formulation. from International Journal of Chemistry May-Aug 2012.

3.      N. Jain et.al., Development And Validation Of RP-HPLC Method For Simultaneous Estimation Of Perindopril erbumine and Amlodipine besilate In Tablet Dosage Forms from Pharma tutor page-37.

4.      Vinjam Swathi et.al., Development and validation of a RP-HPLC method for determination of Perindopril erbumine and Amlodipine besilate in a capsule dosage form American-Eurasian Journal (P) 2008-2012.

5.      M. Saravanamuthukumar et al., Simultaneous Estimation And Validation Of Perindopril erbumine and Amlodipine besilate Tablet Dosage Form By RPHPLC Method Scientific Research 5 (1): 212-215, 2010.

6.      DA Shah, et al., Developed a RP-HPLC method for determination of Perindopril erbumine and Amlodipine besilate in combined tablet dosage form.pharma time Jan- 2014.

7.      Habisiba et al., was reported a validated HPLC and HPTLC methods for simultaneous estimation of Perindopril erbumine and Amlodipine besilate in pure powder and formulations. Der Pharma Chemica, 2014, 6(2):24- 3 (http://derpharmachemica.com/archive.html)

8.      K Arun et al., Developed a reverse phase high performance liquid chromatography method has been reported for the simultaneous estimation of Perindopril erbumine and Amlodipine besilate in formulation. Phrma Times, NO. 19, 2007

9.      Jagadeeswararao V et al., A simple Reverse phase liquid chromatographic method has been developed and subsequently validated for simultaneous determination of fenofibrate in combination International Journal of Chemistry Vol.2, No.8, pp 3012-3025, Nov-Dec 2012.

 

Accepted on 28.06.2019           © RJPT All right reserved

Research J. Pharm. and Tech 2020; 13(5): 2163-2166.