A Systematic Review of Method Development and Validation for Ramipril Analysis using HPLC in Cardiovascular Research

 

Devshree Yashwantbhai Patel, Javesh Kashinath Patil

Department of Pharmaceutical Quality Assurance,

P.S.G.V.P. Mandal’s College of Pharmacy, Shahada, Dist – Nandurbar, 425409.

Associate Professor, Department of Pharmaceutical Quality Assurance,

P.S.G.V.P. Mandal’s College of Pharmacy, Shahada, Dist – Nandurbar, 425409.

 

ABSTRACT:

The field of analytical chemistry comprises many methodologies that are essential in ascertaining the chemical makeup of various substances. This review centers on two widely used techniques: high-performance liquid chromatography (HPLC) and UV-visible spectroscopy. By measuring absorbance using the Beer-Lambert rule, UV-visible spectroscopy can reveal the concentration of absorbing species. It is based on how light and matter interact, with absorption taking place as a result of molecular electronic transitions. HPLC, on the other hand, uses high pressure to separate mixture components according to how well they bind to a stationary phase. Its versatility is attributed to a number of different separation methods, including adsorption, partition, ion exchange, and size exclusion chromatography. In order to guarantee the dependability of analytical techniques, method development and validation are essential. It is necessary to evaluate validation factors, including specificity, accuracy, precision, linearity, and robustness, to ensure that a method is appropriate for the purpose for which it is intended. An analysis of the literature reveals various HPLC techniques for the analysis of ramipril, an ACE inhibitor used to treat heart failure and hypertension. These investigations have examined a range of mobile phases and columns, demonstrating the flexibility of HPLC in pharmaceutical analysis. By blocking the renin-angiotensin-aldosterone system (RAAS), ramipril works by decreasing inflammation and vasoconstriction. An extensive review of analytical chemistry techniques is given in this abstract, with special emphasis on the methods' significance for pharmaceutical analysis and drug development.

 

 

 

INTRODUCTION:

The study of techniques for ascertaining the chemical makeup of material samples is known as analytical chemistry. Information regarding the atomic or molecular species or functional groups of the material can be obtained by a qualitative method. In contrast, a quantitative approach provides numerical data on the proportional amount of one or more of these components¹. Analytical chemistry is measured by a set of powerful concepts and procedures that are applicable to all fields of research and medicine. It is constantly looking for better ways to determine the chemical composition of both natural and synthetic materials. While an analytical method is a specific application of a technique to address an analytical problem, this chemistry discipline is carried out in a large number of laboratories across the Many various methods^2. Analytical techniques are regularly created, enhanced, verified, cooperatively researched, and used. Both qualitative and quantitative analysis are part of the field of analytical chemistry^3,4.

 

Introduction of UV:

UV spectroscopy is a physical method of optical spectroscopy that uses light in the visible, ultraviolet, and near-infrared ranges. It is based on the Beer-Lambert law, which states that the absorbance of a solution is directly proportional to the concentration of the absorbing species in the solution and the path length. Consequently, it may be applied to determine the absorber concentration in a solution for a given path length. Knowing how fast absorbance changes with concentration is important.⁵

 

Principle of UV:

The UV Visible Principle The different spectra that are created when chemical compounds absorb ultraviolet or visible light are the basis of spectroscopy. The relationship between light and matter is the foundation of spectroscopy. When matter absorbs light, it undergoes excitation and de-excitation, which results in the creation of a spectrum. When an electromagnetic wave strikes a medium, a number of processes can occur, including transmission, absorption, reflection, and scattering. The measured spectrum illustrates the interactions of discrete-dimensional objects such as molecules, macromolecules, and atoms at different wavelengths. When the energy difference between the excited and ground states of a molecule equals the frequency of incoming light, absorption takes place. An electronic transition is the process by which an electron is excited from its ground state to its excited state.⁶

 

 

Fig: UV VISIBLE SPECTROSCOPY

HPLC (High Performance Liquid Chromatography):

High-performance liquid chromatography, also referred to as high-pressure liquid chromatography, is a kind of liquid chromatography. The method is widely used in analysis to identify, measure, and separate the constituent parts of a mixture. A more sophisticated kind of column liquid chromatography is called high-performance liquid chromatography (HPLC). Gravity normally drives the solvent through the column, but the high pressures of the HPLC process compress the solvent up to 400 atmospheres, enabling the sample to be divided into different constituents according to variations in relative affinities.⁷

 

The HPLC principle involves injecting the sample's solution into a porous material column (the stationary phase) and then pumping the liquid phase (the mobile phase) through the column at a higher pressure. The solute is adsorbed on the stationary phase according to its affinity for the stationary phase, which follows the separation principle.⁸

The separation process can take four various forms depending on the nature of the stationary phase.

i.    Adsorption chromatography, in which the separation is accomplished through repeated adsorption-desorption processes.

ii. The process of partition chromatography involves dividing the mobile and stationary phases in order to achieve separation.

iii. Anionic surfaces with opposite charges to the sample comprise the separation phase in ion-exchange chromatography; and

iv. Size exclusion chromatography, which separates samples based on their molecular size using a column packed with a substance with precisely controlled pore size.⁹

 

Fig: HPLC (High Performance Liquid chromatography)

 

HPLC METHOD DEVLOPMENT:

 

 

Method Validation:

The analytical method must be validated before the chemical evaluation can be performed. Method validation involves carrying out a series of tests to confirm that an analytical test system is suitable for its intended use and capable of delivering relevant and reliable analytical data. A validation examination tests multiple features of a procedure to see if they can yield accurate information when applied automatically. In order to effectively evaluate method parameters, the validation test should include typical test circumstances such as product excipients. As a result, a technique validation analysis is product-specific.¹⁰

 

1. Specificity: The term "specificity" describes an analytical method's capacity to identify and measure analytes in complicated mixtures. When identifying contaminants and validating identification tests, a specificity inquiry must be carried out. The capacity of HPLC to provide signals free from interference is one of its important properties. The ability to conclusively assess the analyte in the presence of potentially present other compounds is known as specificity, as per ICH recommendations. These are frequently substances such as pollutants, degradants, and matrices.¹¹

 

2. Accuracy: The accuracy of an analytical method is defined as the degree to which test results obtained through this method are close to the actual value. Accuracy is sometimes expressed as a percentage of recovery by assessing known additional amounts of analyte. Applying the methodology to analysed samples with known analyte addition levels allows one to assess the accuracy of an analytical method. The percentage of analyte recovered by the assay is used to calculate the accuracy of test results¹²

 

3. Precision: One way to define accuracy in analytical procedures is "closeness of agreement between a series of measurements obtained from multiple sampling of the same standardised sample under the prescribed conditions".¹³

 

3.1Repeatability: It conveys the accuracy across a short time span with same operational circumstances. i.e., the analysis of duplicates by the analyst using the same methods and instruments.

 

3.2Intermediate Precision: It conveys the accuracy of laboratory variances, such as various days, analysts, and equipment, among other things. Studying affects one at a time is not necessary.

 

3.3Reproducibility: It expresses laboratory precision for the aim of adding procedures to pharmacopoeias (two-way studies, often applied to standardization of method). i.e., As part of the validation procedure for tests used in assay and quantitative impurity testing, precision is examined.

4.Linearity: Linearity is the ability of an analytical procedure to yield a response that is exactly proportionate to the amount or concentration of analyte in the sample.¹⁴

 

5.Range: The difference between the highest and lower concentrations of analyte in an analytical technique with an appropriate degree of linearity, accuracy, and precision is known as the range of the analytical procedure.¹⁵

 

6.Limit of detection: The LOD is the smallest amount of analyte that can be found in the specified experimental conditions.5 It can be calculated using either the 3:1 signal-to-noise ratio or the 3.3:1.5 ratio obtained by dividing the standard deviation of the y-intercepts by the slope of the calibration curve acquired in the linearity test.

 

7.Limit of quantification: A lower-order quantitative quantity, or LOQ, is the smallest quantity of analyte that, within the confines of the experiment, may be precisely and accurately quantified quantitatively. Five

 

The evaluation of this parameter can be done in the same manner as the LOD, but in a 10:1 ratio in each instance.¹⁶

 

8.Robustness: Small adjustments to the optimised technique parameters, such as the mobile phase ratio, flow rate, and detection wavelength, were made in order to conduct the robustness investigation. Retention time and tailing factor are unaffected significantly.¹⁷

 

LITERATURE REVIEW:

	Drug	Authors	Details	Ref.No.
1	Ramipril	Bilal Yilmaz	Mobile Phase:  20 mM phosphate buffer (pH 2.5) containing 0.1% trifluoroacetic acid (TFA)-acetonitrile (50:50, v/v) Column: Ace C18 column (5 µm, 250×4.6 mm	18
2	Ramipril	Vaibhav Rajoriya, Amrita Soni, Varsha Kashaw et. al	Mobile Phase: acetonitrile: methanol (60:40 v/v) Column: ODS C-18 Kromacil (250 mm × 4.60 mm)	19
3	Aspirin + Astovastatin + Ramipril	Nora A. Abdallah1, Amina M. El-Brashy1, Fawzia A. Ibrahim1 and Mohamed I. El-Awad, et. al	Mobile Phase:  0.3% triethylamine (TEA) in 90: 10 an aqueous solution of 0.12 M sodium dodecyl sulfate (SDS): n-propanol, (v/v). The pH was adjusted to 2.5 Column:  Monolithic column	20
4	Ramipril + Amlodipine	Praveen S. Rajput, Amanjot Kaur, Navdeep Kaur Gill, Karan Mittal and Ganti Subrahmanya Sarma. et al	Mobile Phase: Acetonitrile, Sodium phosphate buffer and Methanol in the ratio of 50: 20:25 v/v/v, pH= 6.8 (pH adjusted with OPA). Column:  C18 Column (250 × 4.6 mm i.d. 5 μm particle size),	21
5	Ramipril+ Olmesartan Medoxomil	Deepthi Yada, Divya Yada, T. Rajeshwari, M. Madhavilatha, G. Tulja Rani et al	Mobile Phase: acetonitrile: 0.05 M KH2PO4 pH 3.0 (60:40) Column:  Hypersil C18 (4.6mmx250mm, 5m)	22
6	Ramipril+ Olmesartan Medoxomil.	M. Prasada Rao, M. Srikanth, K. Umamaheswari et al	Mobile Phase: buffer (pH -2.0), methanol and acetonitrile (30:20:50% v/v/v) Column:    ODS C18 column (150 mm x 4.8 mm i.d; particle size 5μm	23
7	Ramipril+Aspirin + Atorvastatin	Rajesh Sharma, Sunil Khanna, and Ganesh P. Mishra et al	Mobile Phase: (A) acetonitrile methanol (65:35) and (B) 10 mM Sodium dihydrogen phosphate monohydrate (NaH2PO4.H2O) buffer and mixture of A: B (60:40 v/v) adjusted to pH 3.0 Column: 25cm×4.6 mm i.d, 5μm particle, C18 column	24

 

Drug Profile:

Ramipril:^25-26

Category: Angiotensin-converting enzyame (ACE) inhibitors.

Molecular formula: C₂₃H₃₂N₂O₅

Chemical name: 2-aza-bicyclo [3.3.0]-octane-3-carboxylic acid.

Molecular weight: 416.5g/mol

Description: White crystalline substance

Melting point: 105°C and 112°C.

Solubility: Poorly water soluble in water, slightly soluble in methanol and very slightly soluble in ethanol

 

Structure:

 

Mechanism of action:

Ramipril prevents the conversion of angiotensin I to angiotensin II by binding to and inhibiting ACE, which inhibits the RAAS system. Angiotensin receptor I (AT1R) and angiotensin receptor II (AT2R), two G-protein coupled receptors, are less activated when plasma levels of angiotensin II decrease.AT1R uses many signaling pathways to mediate oxidative damage, fibrosis, inflammation, and vasoconstriction. These include Gq coupling to the inositol triphosphate pathway, activation of MAP kinases Gi and G12/13 and Ca2+-dependent phospholipases C, A2, and D that aid in the synthesis of eicosanoid compounds, and ultimately activation of the Jak/STAT pathway that promotes cell growth and the synthesis of extracellular matrix components. Reactive oxygen species are produced as a result of AT1R activation and enhanced activity of membrane-bound NADH/NADPH oxidase. Ramipril's renoprotective, antihypertensive, and cardioprotective actions are mediated by decreased activation of this receptor, which also lowers inflammation and vasoconstriction. By activating phosphotyrosine phosphatases that inhibit MAP kinases, blocking Ca2+ channel opening, and promoting the synthesis of cGMP and nitric oxide, which results in vasodilation, AT2R counteracts the actions of AT1R. The Mas receptor, which is activated by Ang (1–7), a subtype of angiotensin generated by plasma esterases from AngI or by ACE2 from AngII produced through a secondary pathway by tonin and cathepsin G, shares these opposing effects. Ang (1–7) also stimulates AT2R, although MasR mediates the majority of its effects.

 

CONCLUSION:

In conclusion, the review gives information about the importance of method development and validation in analytical chemistry, particularly focusing on the HPLC method for Ramipril analysis. The review also focuses on literature review of ramipril which will help us in optimization of chromatographic condition. Also focuses on the significance of validation parameters such as specificity, accuracy, and precision. These parameters are vital in guaranteeing the trustworthiness and efficiency of the analytical approach.

 

REFERENCES:

1.        Skoog D. A, Nieman TA, Holler FJ. Principles of instrumental analysis. 5th ed. London: Stanford University, Saunders College Publications. 1998; 1-4.

2.        Jeffery G.H, Bassett J, Mendham J, And Denney R C., Vogel’s Textbook of Quantitative Chemical Analysis, 5th Ed., Addison Wesley Longman Inc, Singapore. 2001: 4-5.

3.        Erwing G.W., Instrumental Methods of Chemical Analysis, 2nd Ed., Mcgraw Hill Publishing Company Inc. 1960: 1-3.

4.        Skoog D.A., Holler F.J. And Nieman T.A., Principles of Instrumental Analysis, 5th Ed., Thomson Brooks/Cole Asia Pvt. Ltd, Singapore. 2004: 4-7.

5.        Verma G. and Dr. Mishra M.; Development and Optimization of UV-Vis Spectroscopy-A Review. World Journal of Pharmaceutical Research. 2018; 7(11) : 1170-1180

6.        A Review on Uv Visible Spectroscopy. International Journal of Pharmaceutical Research and Applications.  2022; 7(5): 1144-1151 DOI: 10.35629/7781-070511441151

7.        Thammana; review of high-performance liquid chromatography (hplc). RRJPA. 2016; 5(2):22-28

8.        Gita Chawla, Krishna Kr. Chaudhary. A review of HPLC technique covering its pharmaceutical, environmental, forensic, clinical and other application. 2019; 6(2): 27-39

9.        Jacqueline M.R. Belanger J.R. Jocelyn Pare and Michel Sigouin, High Performance Liquid Chromatography (HPLC): Principle and Application. 1997:38-58

10.      Rao, Tentu. Validation of Analytical Methods. Calibration and Validation of Analytical Methods – A Sampling of Current Approaches, edited by Mark Stauffer, IntechOpen, 2018. 10.5772/intechopen.72087.

11.      Thapar, S., Chawla, A., and Gupta, G. K. A concise review on method development and validation parameters. GSC Biological and Pharmaceutical Sciences. 2022; 19(1): 66–77. https://doi.org/10.30574/gscbps.2022.19.1.0136

12.      Patel Kiran, and Patil Javesh. Development and Validation of UV-Spectroscopy and UHPLC Method for Gemfibrozil in Bulk Drug and Pharmaceutical Dosage Form [Data set]. In International Journal of Pharma Research and Technology. 2023;  2(3). Zenodo. https://doi.org/10.5281/zenodo.10039607

13.      Chavan, S. D., and Desai, D. M. Analytical method validation: A brief review. World Journal of Advanced Research and Reviews. 2022; 16(2): 389–402. https://doi.org/10.30574/wjarr.2022.16.2.1165

14.      Patil M A, and Patil J K. Stability Indicating Method Development and Validation of Ziprasidone by using RP-HPLC Method. 2023. https://doi.org/10.5281/zenodo.8394540

15.      Lavanya Chowdary G, Ravisankar P. Int. J. Pharm. Sci. Rev. Res., 2020; 61(2): 1-7

16.      Eserian, J. K., and Lombardo, M. Method validation in pharmaceutical analysis: from theory to practical optimization. Innovations in Pharmacy. 2015; 6(1). https://doi.org/10.24926/iip.v6i1.376

17.      Kapil, Patil and Sunil, Pawar and Patil, Javesh. Development and Validation of RP-HPLC Method for Simultaneous Estimation Of Amoxicillin And Dicloxacillin in Bulk Drug and Capsules. 2014.

18.      Determination of Ramipril in Pharmaceutical Preparations By High-Performance Liquid Chromatography. International Journal of Pharmaceutical Sciences Review and Research. 2010; 1(1): 39–42.

19.      Rajoriya, V., Soni, A., and Kashaw, V. Method Development and Validation of Fast Dissolving Tablet of Ramipril by HPLC Method. International Journal of Pharmacy and Pharmaceutical Sciences. 2016; 8(3): 174–178. https://innovareacademics.in/journals/index.php/ijpps/article/view/9619

20.      Abdallah, N. A., El-Brashy, A., Ibrahim, F., and El-Awady, M. I. Micellar eco-friendly HPLC method for simultaneous analysis of ternary combination of aspirin, atorvastatin and ramipril: application to content uniformity testing. BMC Chemistry. 2023; 17(1). https://doi.org/10.1186/s13065-023-00929-y

21.      Rajput. Simultaneous Estimation of Ramipril and Amlodipine in Bulk and tablet Dosage form by RP-HPLC Method. Journal of Applied Pharmaceutical Science. 2012; https://doi.org/10.7324/japs.2012.2724

22.      Yada, D., Yada, D., Rajeshwari, T., Madhavilatha, M., and Rani, G. T. A New Validated Rp-HPLC Method for The Simultaneous Estimation Of Ramipril And Olmesartan Medoxomil. International Journal of Pharmacy. 2016; 6(1): 177–185. https://www.pharmascholars.com/articles/a-new-validated-rphplc-method-for-the-simultaneous-estimation-of-ramipril-and-olmesartan-medoxomil.pdf

23.      M. Prasada Rao, M. Srikanth, K. Umamaheswari. Simultaneous estimation of Ramipril and Olmesartan Medoximil by RP-HPLC Method.International Journal of Pharmaceutical Chemistry and Analysis. 2017; 4(4): 106-111. DOI: 10.18231/2394-2797.2017.0024

24.      Sharma, R., Khanna, S., and Mishra, G. P. Development and validation of RP-HPLC method for simultaneous estimation of ramipril, aspirin and atorvastatin in pharmaceutical preparations. E-journal of Chemistry. 2012; 9(4): 2177–2184. https://doi.org/10.1155/2012/89169510.

25.      Hitendra Choudhary, Bhagwan Yadav, Pankaj Patel, Prabhat Das, Sujit Pillai. Formulation and Evaluation of Ramipril Fast Dissolving Tablet using Solid Dispersion. Research J. Pharm. And Tech. 2019; 12(8): 3764-3772. Doi: 10.5958/0974-360X.2019.00645.0

26.      Indian Pharmacopoeia, 2007 (Vol. 3). Page No 1648-1469

 

 

 

Received on 17.06.2024       Modified on 24.07.2024

Accepted on 27.08.2024   ©Asian Pharma Press All Right Reserved

Asian J. Pharm. Ana. 2024; 14(3):180-184.