Analytical Methodologies for Ranolazine estimation: A brief review

Jitendra Yadav*, Dolly Dewangan, Kamraj, Krity Gupta, Manish Kumar Sahu,

Parimal Verma, Prashant Kumar Sahu, Shweta Sinha, Aakanksha Sinha, S. J. Daharwal

University Institute of Pharmacy, Pt. Ravishankar Shukla University, Raipur (C.G).

*Corresponding Author E-mail: jy7288096@gmail.com

ABSTRACT:

Ranolazine, a piperazine derivative, is an anti-anginal drug used to treat cardiac ischemia. In myocardial ischemia, it works by altering sodium-dependent calcium channels, so avoiding calcium overload, which results in heart ischemia. When taken in conjunction with other treatments such as beta-blockers, nitrates, calcium channel blockers, antiplatelet medication, lipid-lowering medicines, ACE inhibitors, and angiotensin receptor blockers, ranolazine is helpful for treating chronic angina. A new anti-ischemic drug called ranolazine inhibits reverse mode sodium-calcium exchange, which lowers diastolic calcium accumulation by suppressing late INa to prevent cellular sodium overload. By directly addressing myocardial ischemia, this technique can raise coronary blood flow and diastolic tone. The several analytical techniques that have been documented for the determination of ranolazine in synthetic mixtures are represented in this review article. Chromatographic techniques were reported, including GC, LC-MS, RP-HPLC, HPTLC, HPLC, and LC-MS/MS.

KEYWORDS: Ranolazine, HPTLC, HPLC, LC-MS.

1. INTRODUCTION:

Ranolazine {(+N-(2, 6-dimethylphenyl)-4(2-hydroxy-3-(2-methoxyphenoxy)-propyl)-1-piperzine acetamide dihydrochloride)} s an active piperazine derivative that comes in oral and intravenous forms, and it was patented in 1986.¹Piperazines are chemical compounds made up of a six-membered ring with two nitrogen atoms positioned opposite each other. Clinical management of chronic angina pectoris involves the use of ranolazine.²The anti-ischemic and anti-anginal actions of ranolazine are independent of drops in heart rate or blood pressure. It is anticipated that ranolazine will lessen sodium influx into ischemic myocardium cells by lowering the late sodium current. Therefore, it is suggested that ranolazine decrease calcium uptake indirectly through the sodium/calcium exchanger. For patients whose response to other anti-anginal medications has been insufficient, the medication may be used in conjunction with them. In order to treat persistent angina, extended-release ranolazine was recently approved in the US.^3-6

By blocking late sodium channels and avoiding intracellular calcium excess, ranolazine, a second-line anti-anginal medication, reduces angina symptoms, myocyte relaxation, and diastolic tension. Because of its distinct inhibitory action in the late phase of the inward sodium current in cardiac myocyte, which lowers myocardial oxygen demand without appreciably altering heart rate or blood pressure, ranolazine was selected for our study over other second-line medications like ivabradine or nicorandil. Patients with refractory angina who are not responsive to other anti-anginal treatments may be candidates for ranolazine.⁷

Fig 1: Structure of Ranolazine

PROPERTIES:

1. Physiochemical properties:

Ranolazine is a white to off-white solid that dissolves in natural solvents like methanol and dichloromethane, is only weakly soluble in ethanol, acetonitrile, acetone, and tetrahydrofuran, and only weakly soluble in ethyl acetate, iso-propanol, toluene, and ethyl ether. It is also very weakly soluble in water. For ROZ, Pka is 2.2 and the partition coefficient (log p) is 2.07. ⁸ In methanol, ranolazine dissolves easily. Ranolazine pKa value of 13.6 indicates that it is a strong base due to its six-membered piperazine ring. The melting point of ranolazine is 122-124 °C.⁹

2. Pharmacological properties:

Ranolazine is thought to work by inhibiting the late inward sodium current in heart cells. Additionally, ranolazine suppresses the potassium current (IKr), the rapidly activating component of the delayed rectifier. Although ranolazine has anti-anginal properties, it has no discernible effect on blood pressure or heart rate. Ranolazine has shown anti-arrhythmic effect in preclinical and clinical trials and does not seem to have the pro-arrhythmic activity usually associated with medications that inhibit IKr, even if it slightly lengthens the QTc interval. In individuals with non-ST-elevation acute coronary syndromes, chronic stable angina, or diabetes mellitus, ranolazine ER enhanced glycaemic management.¹⁰

Mechanism of action:

Ranolazine inhibits late INa in cardiac myocyte of dogs and guinea pigs, showing dependency on frequency, voltage, and concentration. It mitigates sodium overload, improves diastolic tone, and enhances coronary blood flow without altering heart rhythm. It lengthens action potential and QT interval by 2–5 ms, decreases post-ischemic contracture in rabbits, and does not promote repolarization dispersion or early afterdepolarizations, which reduces the risk of Torsade de pointes. The drug also lowers intracellular calcium and salt levels, effectively reducing oxygen consumption while not impacting blood pressure or heart rate. In the MARISA trial, ranolazine extended exercise duration in angina patients without adverse hemodynamic effects and promotes glucose oxidation and cardiac strengthening by enhancing fatty acid oxidation and delaying action potential through potassium current suppression.¹¹

Fig 2: Mechanism of action of Ranolazine

Clinical effects of ranolazine:

In patients with chronic stable angina, the effectiveness of sustained-release ranolazine was evaluated in two phase three trials, MARISA and CARISA. In the 191-patient crossover research MARISA, ranolazine mono-therapy was found to significantly improve exercise performance and lower the frequency of angina during testing (52% against 70% of placebo). Likewise, over a 12-week period, CARISA, which included ranolazine in addition to pre-existing anti-anginal medication, showed improvements in activity duration and a decrease in angina attacks from 4.5 to 2.1 per week, as opposed to 3.3 for a placebo. Notably, neither blood pressure nor heart rate changed significantly during these impacts. Furthermore, the effect of ranolazine on clinical outcomes in acute coronary syndromes was investigated in the MERLIN TIMI-36 experiment.^12-13

Pharmacology:

Pharmacodynamic:

Ranolazine is a dihydrochloride compound that inhibits sodium and potassium ion channel currents at therapeutic levels. During cardiac repolarization, it particularly impacts the late phase of the inward sodium current. Increased sodium-calcium exchange in pathological situations leads to higher cytosolic calcium concentrations, which impede left ventricular relaxation after ischemia and reperfusion. Ventricular tachycardia is more likely as a result of this calcium excess since it also increases left ventricular diastolic wall tension, lowers myocardial blood flow, and impairs myocardial electrical stability.^14-15

Other actions of ranolazine include suppression of fatty acid oxidation and binding to adrenergic receptors. In animal experiments, it functions as an a1- and b1-adrenergic antagonist without appreciably impairing heart rate or contractility at rest or during exercise. Nonetheless, stress has been found to alter heart rate and rate pressure product in human beings. At therapeutic levels, the original theory that its main anti-ischemic action was fatty acid oxidation inhibition has been shown to be false. Inhibiting the late phase of the inward sodium current in cardiac cells is linked to its anti-anginal activity; this lowers intracellular calcium excess and diastolic dysfunction. It is necessary to conduct additional research on how it affects human heart rate. ^16-17

Pharmacokinetics:

Ranolazine was first studied using immediate-release capsules; however, an extended-release formulation was created because of its short half-life and inconsistent absorption. A steady state is reached within three days of twice-daily dosing, with a steady-state half-life of seven hours, and peak plasma concentration happens two to six hours after delivery. Ranolazine is mostly eliminated by the kidneys (75%), after being substantially processed by the liver by cytochrome P450. Its pharmacokinetics are unaffected by age, sex, or congestive heart failure. ¹⁸It is given orally as tablets, usually 500 mg twice a day at first, then as tolerated, up to 1000 mg twice a day. The plasma reaches its highest level in 2 to 5 hours, the half-life is 7 hours, and the steady-state concentration is reached in 3 days. Since CYP3A4 and CYP2D6 are the primary enzymes in the liver that metabolize ranolazine, it is not recommended to use inhibitors of these enzymes. By blocking fatty acid metabolism, the medication increases glucose oxidation and decreases the generation of lactic acid. These are its two primary targets and routes of action.¹⁹

Interactions/Precautions:

CYP3A4 is primarily responsible for ranolazine's hepatic metabolism, with CYP2D6 playing a role. Strong CYP3A4 inhibitors such as diltiazem, HIV protease inhibitors, macrolide antibiotics (like clarithromycin), certain antifungals (like ketoconazole), and grapefruit products should not be taken with it. On the other hand, with the exception of some antipsychotics and some tricyclic antidepressants, co-administration with CYP2D6 inhibitors usually does not necessitate dosage modifications.²⁰

Importance of Analytical Method Development and Validation:

Validating analytical techniques is crucial for regulatory requirements, sound research, and quality control. CFR 311.165c requires that test techniques be accurate, sensitive, specific, and repeatable. Scientists should employ high-quality science to demonstrate the method's accuracy, sensitivity, specificity, and repeatability. Even though it is costly and time-consuming at first, analytical method validation increases productivity, eliminates repetitive tasks, and boosts developer and customer confidence.^21-22

Development and Validation of HPLC Methods for Analytical Applications:

In 1903, Russian botanist M.S. Tswett developed High Performance Liquid Chromatography (HPLC), a popular analytical technique used in more than 85% of generic pharmaceuticals. Chemical separation, identification, and quantification based on migration rates between stationary and mobile phases are achieved by injecting a sample solution into a porous column and forcing a liquid through it at high pressure. Effective HPLC requires advanced technology such as computer modeling and limited experimental runs for optimization and performance evaluation. ^23-24

Table 1: HPLC based analytical method development for ranolazine

Title	Method	Stationary phase	Mobile phase	Wavelength	Reference
Validated HPLC Method for Determining Related Substances in Compatibility Studies and Novel Extended Release Formulation for Ranolazine	HPLC	Supelcosil C 18, (250×4.6 mm, 5 µm) column	combination of Phosphate buffer pH 7.0 and Methanol in ratio of 350:650	220 nm	25
A validating stability indicating reversed phase high performance liquid chromatography method of ranolazine dihydrochloride and characterization of its degradation products	HPLC	Novapak C18 column	Methanol-acetonitrile-phosphate buffer (pH 3.6; 6.3 mM) (4:3:3, v/v/v)	220 nm	26
Development and validation of a new analytical method for the determination of related component and assay of ranolazine in bulk and pharmaceutical dosage form by LC	HPLC	-	Buffer- acetonitrile (90:10 v/v) and Acetonitrile- water (90:10 v/v)	-	27
Determination of Ranolazine in human plasma	HPLC	Agilent ZORBAX C18 column	Acetonitrile:0.1% formic acid (90:10)	-	28
Estimation of Ranolazine HCL in Tablet Dosage form	RP-HPLC	Inertsil ODS C18	Buffer: Acetonitrile (60:40) pH adjust with tri-ethylamine	224 nm	29
Estimation of Ranolazine in tablet dosage form	RP- HPLC	X-terra C18 column	Sodium dihydrogen phosphate buffer (pH adjust to 5 with dilute ortho-phosphoric acid) : Acetonitrile (600:400)	210 nm	30
Estimation of Ranolazine in Bulk and Tablet Dosage Form	RP-HPLC	ODS C18 column	Ammonium acetate buffer pH-4: Acetonitrile: Methanol (30:50:2)	200 nm	31
Estimation of Ranolazine in bulk and Pharmaceuticals formulation	RP-HPLC	X-terra RP18 column	Sodium dihydrogen phosphate buffer pH adjust to 5 with dilute ortho-phosphoric acid: Acetonitrile (600:400	225 nm	32
Estimation of Ranolazine	RP-HPLC	Agilent Eclipse XDB C18 column	Phosphate buffer pH 3.5 : Acetonitrile 65:35 (v/v)	272 nm	33
Method for Ranolazine dihydrochloride and its degradation product	RP-HPLC	C18 column	Methanol: Acetonitrile : phosphate buffer (pH 3.6,6.3 mM) (4 : 3 :3, V/V)	220 nm	34
Determination of Ranolazine drug substance and drug product	RP-UPLC	Acquity BEH RP18 Column	Monobasic sodium buffer : acetonitrile	-	35
Quantitation of ranolazine in human plasma	U-HPLCMS/MS	BEH C18 column	Acetonitrile aqueous ammonium acetate solution (40:60, V/V)	-	36
Ranolazine in bulk and marketed formulation	HPLC-UV	-	Methanol: 0.5% tri- ethylamine, pH6 with ortho-phosphoric acid (75:25)	271 nm	37

Method Development and Validation of LC, LC-MS and GC Techniques for Quantitative Analysis:

In pharmaceutical analysis, liquid chromatography (LC), particularly high-performance liquid chromatography (HPLC), is a crucial separation method that focuses on drug assay and purity testing. The primary objective of technique development is to achieve adequate separation between the target analyte and impurities and degradation products. A highly efficient technique for tracking, describing, and identifying contaminants in pharmaceutical analysis, as well as in the analytical development and quality control of drug substances and dosage forms, is LC-MS (Liquid Chromatography–Mass Spectrometry), which combines liquid chromatography and mass spectrometry.³⁸

Gas chromatography (GC) is an analytical technique initially developed for gases and volatile components, significantly advanced by pioneers Martin and Synge, and later James and Martin. Derivatization or pyrolysis GC can be used to analyze nonvolatile samples, while it enables the separation and analysis of gaseous samples, liquid solutions, and volatile solids. ³⁹

Table 2: LC, LC-MS and GC based analytical method development for ranolazine

Title	Method	Stationary phase	Mobile phase	wavelength	Reference
Determination of Ranolazine H CL in bulk and dosage form	LC	HiQ Sil C18 H S	Methanol: water (99:1 %, V/V)	273 nm	40
Quantitation of Ranolazine in rat plasma	LC	Nova-Pak C18 column	Acetonitrile: water: formic acid: 10% n butyl-amine (70:30:0.5:0.08, v/v/v/v)	-	41
Estimation of Ranolazine in Human Plasma	LC	Zorbax extend C18 column	Methanol–10mM acetate (60:40 v/v, pH 4.0)	-	42
Semi preparative resolution of ranolazine enantiomers	LC	Cellulose tris (3 ,5di-methyl phenyl-carbonate) Chiral stationary phases	Methanol	-	43
Determination of Related Component and Assay of Ranolazine	LC	C18 column	-	210 nm	44
Quantitation of Ranolazine in rat plasma	LC	C18 column	-	-	45
Estimation of Ranolazine an d eleven phase I metabolites	LC-MS	Source- Finni gan MAT TSQ 700	-	-	46
Determination of ranolazine in rat plasma	LC-MS	C18 column	Methanol:10 mM ammonium acetate (76 : 24 V/V)	-	47
Estimation of ranolazine in dog urine	LC-MS	-	-	-	48
Analysis of Ranolazine an d Di-methyl ranolazine	LC-MS/MS	Chiracel ODH Column	-	-	49
Quantitation of ranolazine and its three metabolites	LC-MS/MS	Gemini C18 column	Methanol: 5 mM ammonium acetate	-	50
Estimation of Ranolazine in human plasma	LC-MS/MS	Peerless Cyano-column	Methanol water containing formic acid (1.0%, v/v) (65:35, v/v)	-	51
Determination of residual solvents in ranolazine by headspace gas chromatography	GC	HP-INNOWAX column	-	-	52

Method Development and Validation of HPTLC Techniques for Quantitative Analysis:

A more sophisticated and automated variation of traditional Thin-Layer Chromatography (TLC), sometimes referred to as planar chromatography, is HPTLC, or High-Performance Thin-Layer Chromatography. In contrast to TLC, it incorporates notable advancements in methodology and technology, such as process automation that improves scanning and sample application precision. ⁵³

Table3: HPTLC based analytical method development for ranolazine

Title	Method	Stationary phase	Mobile phase	Wavelength	Reference
Stability indicating method development and validation of Ranolazine hydrochloride in bulk and tablet form by HPTLC	HPTLC	silica gel aluminium	Chloroform : Methanol : Toluene (5 : 1 : 1 v/v/v)	273 nm	54
High performance thin layer chromatography estimation of ranolazine	HPTLC	Aluminium plates pre-coated with Silica gel G 60 F254	Methanol : 10mM ammonium acetate solution (6:4 V/V)	271 nm	55
Determination of Ranolazine in tablet formulation by High performance thin layer chromatography- Mass spectrometry using reflectance scanning densitometry	HPTLC-MS	_	Butanol–Acetic acid–water (6:2:2 v/v)	270 nm	56

Development and Validation of a UV–Spectrophotometric Analytical Methods:

UV spectroscopy is a method that uses ultraviolet and visible light to analyze the chemical structure, dynamics, and composition of molecules. In addition to identifying contaminants and organic waste, it enables the sensitive and rapid investigation of ecosystem activity. The sensitivity, specificity, and real-time process management of UV spectroscopy are enhanced by the use of nanotechnology in pharmaceutical analysis, biomedical applications, online monitoring systems, and environmental monitoring.^{57- 58}

Table 4: UV based analytical method development for ranolazine

Title	Method/ instrument	Sample	Solvent	Wavelength	Reference
Spectrophotometric method for the estimation of ranolazine in bulk and pharmaceutical formulation	Shimadzu UV/visible double beam spectrophotometer (Model 2450)	Bulk and formulation	distilled water	447 nm	59
Nano-drop Spectrophotometric method development and validation for estimation of ranolazine in their bulk	Nano-drop spectrophotometer ND-100	Bulk	distilled water	272 nm	60
Development and validation of UV spectrophotometric area under curve method for Quantitative estimation of Ranolazine in API and Tablet Formulation	Area under curve method Shimadzu UV 1800 with UV Prob Software	API and Tablet	Methanol and distilled water	261-281 nm	61
Development and validation of analytical methods for Quantitative estimation of ranolazine	UV-VIS spectrophotometric	Bulk and Tablet	0.05 N HCl, potassium ferricyanide	270, 449	62
Development and validation of UV Spectrophotometric method for the estimation of ranolazine in bulk drug and pharmaceutical formulation	UV spectrophotometer (Shimadzu UV-1800)	Bulk drug and formulation	Methanol	272 nm	63
Derivative Spectrophotometric Methods For the analysis and Stability Studies Of Ranolazine in bulk and Dosage Forms	Shimatzu UV- 1800EN240V, double beam, (Kyoto, Japan).	Bulk and dosage form	Methanol	278 nm, 283nm	64
Novel analytical method development for some amide group containing drugs using Bougainvillea spectabilis bract extracts	UV-visible spectrophotometer	Bulk	Methanol	418nm	65

Table 5: Various drug based simultaneous estimation of Ranolazine

Title	Method	Data	Wavelength	Reference
First order derivative Spectrophotometric estimation of ranolazine and Metformin Hydrochloride in synthetic mixture	First order derivative method	Solvent-water	232.86, 249.29 (Zero crossing point for Metformin hydrochloride and ranolazine	66
Novel UV-spectrophotometric and RP-HPLC method development and validation of simultaneous estimation of ranolazine and Metformin HCL: A statistical analysis	UV and RP-HPLC	UV data- Methanol RP-HPLC data- S.P.-C18 (250 mm x 4.6 mm, 5µm) M.P.- acetonitrile, methanol and 0.05 M ammonium formate buffer (PH-6) (25:60:35)	UV data- Metformin HCL- 251 nm Ranolazine- 237 nm RP-HPLC data- 228 nm	67
Development of a UV visible spectrophotometric method for simultaneous estimation of Ranolazine and Metoprolol	UV	Solvent- 0.1 N HCl	Ranolazine- 272nm Metoprolol- 242nm	68
Simultaneous estimation of Ranolazine and Dronederone in bulk and pharmaceutical dosage forms.	HPLC	Mobile phase-0.02N NH2PO4 buffer (pH4) Acetonitrile (50 :50 V/V) Stationary phase- ODS column	282nm	69

CONCLUSION:

In order to evaluate ranolazine in different forms and biological fluids, the paper addresses the need for ongoing development of precise, sensitive, and efficient analytical techniques. In addition to providing details on the Pharmacodynamics, pharmacokinetics, and mechanism of action of ranolazine, it highlights the review of the many analytical techniques used for its testing. There are several different ways for determining ranolazine in pharmaceutical formulations, but spectrometric approaches are the most straightforward and cost-effective. It is well known that, in comparison to more sophisticated detection methods RP-HPLC, HPLC, UV, LC, LC-MS, HPTLC, HPLC-UV and GC offer accurate results at reduced costs. It is believed that this information will be very helpful for future research on ranolazine formulation development, analytical method development and quality control.

FUTURE PROSPECTS:

Ranolazine drug has been studied in many analytical method but we see that it can studied in some analytical method like ion charge method, Chemo-metric analysis methods etc.

REFERENCE:

1. Rousseau MF, Pouleur H, Cocco G, Wolff AA. Comparative efficacy of ranolazine versus atenolol for chronic angina pectoris. The American Journal of Cardiology. 2005 Feb 1;95(3):311-6.

2. Djamgoz MB. Ranolazine: a potential anti-metastatic drug targeting voltage-gated sodium channels. British Journal of Cancer. 2024 May 18; 130(9):1415-9.

3. Nalawade V, Gide P, Nunnavare G, Phale M. Development and validation of a LLE-HPLC method for the determination of ranolazine in human plasma. Int. J. Ph. Sci. 2010 Jan;2(1).

4. Markus J. Clinical pharmacokinetics of ranolazine; Clin. P kinetics. 2006; 45(5):469-91.

5. Stanley WC. Ranolazine: new approach for the treatment of stable angina pectoris. Expert Review of Cardiovascular Therapy. 2005 Sep 1; 3(5):821-9.

6. Extended-release ranolazine [Ranexa; CV Therapeutics] has been launched in the US for the treatment of chronic angina, Inpharma, Adis International Publisher, pp18-18(1)-2006.

7. Ling H, Fu S, Xu M, Wang B, Li B, Li Y, Liu X, Zhang X, Wang Q, Li A, Liu M. Ranolazine for improving coronary microvascular function in patients with nonobstructive coronary artery disease: a systematic review and meta-analysis with a trial sequential analysis of randomized controlled trials. Quantitative Imaging in Medicine and Surgery. 2024 Jan 9;14(2):1451.

8. Patil P, Rangari S, Patil P. Research on analytical methods of analysis of Ranolazine: A Review. Himalayan Journal of Health Sciences (HJHS). 2020:30-40.

9. Patel Vishakha. D. Raj Hasumati, Gheewala Nirav, A Review on Analytical Methods for Ranolazine determination in synthetic mixture; Pharma Tutor; 2016; 4(4); 28-31

10. Keating GM. Ranolazine: a review of its use in chronic stable angina pectoris. Drugs. 2008 Dec; 68(17):2483-503

11. Hasenfuß G, Maier LS. Mechanism of action of the new anti-ischemia drug ranolazine. Clinical Research in Cardiology. 2008 Apr; 97(4): 222-6.

12. Chaitman BR, Pepine CJ, Parker JO, Skopal J, Chumakova G, Kuch J, Wang W, Skettino SL, Wolff AA, Combination Assessment of Ranolazine In Stable Angina (CARISA) Investigators, Combination Assessment of Ranolazine In Stable Angina (CARISA) Investigators. Effects of ranolazine with atenolol, amlodipine, or diltiazem on exercise tolerance and angina frequency in patients with severe chronic angina: a randomized controlled trial. JAMA. 2004 Jan 21; 291(3): 309-16.

13. Chaitman BR, Skettino SL, Parker JO, Hanley P, Meluzin J, Kuch J, Pepine CJ, Wang W, Nelson JJ, Hebert DA, Wolff AA. Anti-ischemic effects and long-term survival during ranolazine monotherapy in patients with chronic severe angina. Journal of the American College of Cardiology. 2004 Apr 21; 43(8): 1375-82.

14. Tani M, Neely JR. Role of intracellular Na+ in Ca2+ overload and depressed recovery of ventricular function of reperfused ischemic rat hearts. Possible involvement of H+-Na+ and Na+-Ca2+ exchange. Circulation research. 1989 Oct;65(4):1045-56.

15. Hale SL, Shryock JC, Belardinelli L, Sweeney M, Kloner RA. Late sodium current inhibition as a new cardioprotective approach. Journal of Molecular and Cellular Cardiology. 2008 Jun 1; 44(6): 954-67.

16. Létienne R, Vié B, Puech A, Vieu S, Le Grand B, John GW. Evidence that ranolazine behaves as a weak β1-and β2-adrenoceptor antagonist in the rat cardiovascular system. Naunyn-Schmiedeberg's Archives of Pharmacology. 2001 Apr;363(4):464-71

17. Bairey Merz CN, Handberg EM, Shufelt CL, Mehta PK, Minissian MB, Wei J, Thomson LE, Berman DS, Shaw LJ, Petersen JW, Brown GH. A randomized, placebo-controlled trial of late Na current inhibition (ranolazine) in coronary microvascular dysfunction (CMD): impact on angina and myocardial perfusion reserve. European Heart Journal. 2016 May 14; 37(19): 1504-13.

18. Jerling M. Clinical pharmacokinetics of ranolazine. Clin Pharmacokinet. 2006; 45: 469–491.

19. McCormack JG, Barr RL, Wolff AA, Lopaschuk GD. Ranolazine stimulates glucose oxidation in normoxic, ischemic, and reperfused ischemic rat hearts. Circulation. 1996 Jan 1; 93(1):135-42.

20. Fihn SD, Gardin JM, Abrams J, Berra K, Blankenship JC, Dallas AP, Douglas PS, Foody JM, Gerber TC, Hinderliter AL, King SB. 2012 ACCF/AHA/ACP/AATS/PCNA/SCAI/STS guideline for the diagnosis and management of patients with stable ischemic heart disease: a report of the American College of Cardiology Foundation/American Heart Association task force on practice guidelines, and the American College of Physicians, American Association for Thoracic Surgery, Preventive Cardiovascular Nurses Association, Society for Cardiovascular Angiography and Interventions, and Society of Thoracic Surgeons. Journal of the American College of Cardiology. 2012 Dec 18; 60(24):e44-164.

21. Lavanya G, Sunil MM, Eswarudu MM, Eswaraiah MC, Harisudha K, Spandana BN. Analytical method validation: an updated review. International Journal of Pharmaceutical Sciences and Research. 2013 Apr 1; 4(4):1280..

22. Pranshu T, Singh RP, Vikash J. Validation: A critical parameter for quality control of pharmaceuticals. Journal of Drug Delivery and Therapeutics. 2012; 2(3):34-40.

23. Ravisankar, P., S. Gowthami, and G. Devlala Rao. A review on analytical method development. Indian Journal of Research in Pharmacy and Biotechnology. 2014; 2(3): 1183

24. Vidushi Y, Meenakshi B, Bharkatiya M. A review on HPLC method development and validation. Res J Life Sci, Bioinform, Pharm Chem Sci. 2017; 2(6): 178.

25. Suresh Babu VV, Sudhakar V, Murthy TE. Validated HPLC method for determining related substances in compatibility studies and novel extended release formulation for ranolazine. J Chromatograph Separat Techniq. 2014; 5(209): 2.

26. Laha TK, Padhy DR, Sen S. A validated stability indicating reversed phase high performance liquid Chromatographic method of ranolazine dihydrochloride and Characterization of its degradation products. International Journal of Pharmacy and Pharmaceutical Sciences. 2013; 5(1): 61-6.

27. Madhavi A, Subba Rao DV, Srinivasu P, Naidu A. Development and validation of a new analytical method for the determination of related components and assay of ranolazine in bulk drug and pharmaceutical dosage forms by LC. Chromatographia. 2009 Jul; 70(1): 333-8

28. Bai S, Gao H, Qu H, Liang Y, Li Y, Zheng Z, Wang X, Hao G. Quantitative determination of Ranolazine in human plasma by high performance liquid chromatography-tandem mass spectrometry. Northwest Pharmaceutical Journal. 2011(4):249-51. Patel R.C, Rathod D.K, Patel P.R, Patel V.S, Estimation of Ranolazine Hydrochloride by Spectrophotometric and RP-HPLC in tablet dosage forms. International Journal of Pharmaceutical and Applied Sciences. 2010; 1(2): 79-83

29. G.L, K.H, Agarwal N, Development and validation of stability indicating RP-HPLC method for estimation of Ranolazine in bulk and tablet dosage form. International Research Journal of Pharmaceutical and Applied Sciences. 2012; 2(5): 217-223

30. Soujanya A, K.V.S, Puttagunta S, Method Development and Validation for estimation of Ranolazine in its bulk and tablet dosage form. International Journal of Inventions in Pharmaceutical Sciences. 2013; 1(4): 363-369

31. Ganji R, D. R, G. S, Jayapal G, P. R, Srilakshmi V, Development and Validation of stability indicating RP-LC method for estimation of Ranolazine in bulk and its Pharmaceutical formulation., American Journal of analytical Chemistry, 2012, 3, 378-384

32. Boovizhikannan T, Palanirajan V, Estimation of Ranolazine using Reverse phase High Performance liquid Chromatography Technique. American Journal of Pharma Tech Research. 2012; 2(6),

33. Laha T, Padhy D, Sen S, A Validated stability indicating reversed phase high performance liquid chromatography method of Ranolazine Dihydrochloride and characterization of its degradation products. International Journal of Pharmacy and Pharmaceutical Sciences. 2013; 5(1): 61-66

34. Malati V, Reddy A.R, Mukkanti K, Suryanarayana M.V, A novel reversed phase stability indicating RP-UPLC method for the quantitative determination of fifteen related substances in Ranolazine drug substance and drug product. J. Talanta. 2012; 97: 563-373.

35. Tan Q, Zhu R, Li H, Fang P, Yan M, Zhang Q, Peng W, Development and Validation of a,sensitive,U HPLC-MS/MS, method, with electrospray ionization for quantitation of Ranolazine in human plasma: Application to a clinical pharmacokinetic study. Journal of Chromatography B, 2012; 901: 119-124

36. Parvatha reddy S, Desam N, Nuthalapati M, B.T, Development and Validation of HPLC and UV methods for estimation of Ranolazine in Bulk and Marketed formulation. International Journal of Innovative Pharmaceutical Sciences and Research. 2014; 2(5): 1042-1058

37. Ermer J, Vogel M. Applications of hyphenated LC‐MS techniques in pharmaceutical analysis. Biomedical Chromatography. 2000 Oct; 14(6): 373-83

38. Bhardwaj SK, Dwivedi K, Agarwal DD. A review: GC method development and validation. Int J Anal Bioanal Chem. 2016; 6(1):1-7

39. Sharma T, Moitra S.K, Si.S.C, Sankar D.G, Stability indicating LC method for the determination of Ranolazine Hydrochloride in the bulk drug and in pharmaceutical dosage form. International Journal of Pharmacy and Pharmaceutical Sciences. 2011; 3(4): 327-332

40. Lei T, Juanjuan J, Yiling H, Lu H, Hong L, Yishi L. Sensitive quantification of Ranolazine in human plasma by liquid chromatography-tandem mass spectrometry with positive electrospray ionization. Journal of Chromatography B, 2007; 846(1-2): 346-350

41. Zhao L, Li H, Jiang Y, Piao R, Li P, Gu J. Determination of Ranolazine in human plasma by liquid chromatographic-tandem Mass Spectrometric assay. Journal of Chromatographic Science. 2008; 46(8): 697-700

42. Luo X, Zhai Z, Wu X, Shi Y, Chen L, Li Y. Analytical and semi-preparative resolution of Ranolazine enantiomers by, liquid, chromatography using polysaccharide chiral stationary phases. Journal of Separation Science. 2006; 29(1): 164-171

43. A.M, Venkata S.D, P.S, A.N, Development and Validation of a new analytical method for the Determination of related components and assay of Ranolazine in bulk drug and pharmaceutical dosage forms by LC., J. Chromatographia, 2009; 70(1): 333-338

44. Liang Y, Xie L, Liu XD, Chen WD, Lu T, Xiong Y, Wang Gj, Simple, sensitive and rapid liquid chromatography/atmospheric pressure chemical ionization mass spectrometric method for the quantitation of Ranolazine in rat plasma., Rapid Commun Mass Spectrum. 2006; 20(4): 523-52

45. Herron W.J, Eadie J, Penman A.D, Estimation of Ranolazine and eleven phase I metabolites in human plasma by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry with selected-ion monitoring. Journal of Chromatography A. 1995; 712(1): 50-60

46. Zhong J, Liu X.Q, Chen Y, Zhao X.P, Wang Y.S, Wang G.J. Determination of Ranolazine in rat plasma by liquid chromatography-electrospray ionization mass spectrometry., J. Chromatographia. 2006; 63(3-4): 123-127

47. Tang JY, Wang w, Li L, Zhang CG, Wang DM, Sun L, Analysis of primary metabolites of Ranolazine in dog urine by LC-MS(n)., Yao Xue Xue Bao. 2012; 47(8): 1031-1038

48. Simões RA, Barth T, Bonato PS. Enantioselective analysis of ranolazine and desmethyl ranolazine in microsomal medium using dispersive liquid–liquid microextraction and LC–MS/MS. Bioanalysis. 2013 Jan 1;5(2):171-83.

49. Wang Y, Chen X, Sun Z, Yang Y, Liu W, Zhong D, Development and Validation of a sensitive LC MS/MS assay for simultaneous quantitataion of Ranolazine and its three metabolites in human plasma., Journal of Chromatography B analyt Technology Biomed Life Sciences. 2012; 10(6): 889 890

50. Bhaumik U, Ghosh A, Sarkar A, BoseA, Selvan P, Sengupta P, Chakraborty U, Ghosh D, Pal T, Determination of Ranolazine in human plasma by LC-MS/MS and its application in bioequivalence study., Journal of Pharmaceutical and Biomedical Analysis. 2008; 48(5): 1404-1410

51. Ya-Lin Ye, Xue-Mei Yang. Determination of Residual Solvents in Ranolazine By Headspace Gas Chromatography. Journal of Southern Medical University. 2008; 28(1): 134-135.

52. Patel RB, Patel MR, Batel BG. Experimental aspects and implementation of HPTLC. InHigh-performance thin-layer chromatography (HPTLC) 2011 (pp. 41-54). Springer, Berlin, Heidelberg.

53. A. Khedkar, et al. Stability Indicating Method Development and Validation of Ranolazine Hydrochloride in Bulk and Tablet dosage form by HPTLC. International Journal of Pharmaceutical and Clinical Research. 2015; 7(1): 77-83.

54. Chakraborty R, Pal K, Shaiba M, Sangepu N, Sridevi P. High performance thin-layer chromatographic estimation of ranolazine. Res. J. Pharm. Biol. Chem. Sci. 2010; 4: 152-7.

55. Abburu SR, Alapati ML, Dadhich AS, Rao MK. Determination of ranolazine in tablet formulations by high-performance thin-layer chromatography—mass spectrometry using reflectance scanning densitometry. JPC-Journal of Planar Chromatography-Modern TLC. 2016 Jun; 29(3): 190-4

56. Nnachi AB. Advances in UV spectroscopy for monitoring the environment. RIJBAS. 2024; 3(1): 82.

57. Maurya RK, Kumar GP, Singh G, Malik J, Singh SP, Tiwari A. Recent developments in UV-Spectroscopy techniques for quantitative analysis in pharmaceutical dosage forms.

58. Sharma A, Prakash D, Singh SK. Development and validation of UV spectrophotometric method for the estimation of ranolazine in bulk drug and pharmaceutical formulation. Int J Chem Tech Res. 2010 Oct; 2(4): 1945-8.

59. Singh RK, Patel PS, Malik P, Singh TP. Nanodrop Spectrophotometric method development and validation for estimation of Ranolazine in Their Bulk. International Journal of Pharmaceutical Sciences and Research. 2011 Apr 1;2(4):985.

60. Ugale JB, Mulgund SV. Development and Validation of UV Spectrophotometric Area under cure method for Quantitative Estimation of Ranolazine in API and Tablet Formulation. World Journal of Pharmaceutical Research. 2015 Mar 16;4(5):2665-72.

61. Nishith, C., Development and validation of analytical methods for quantitative estimation of ranolazine. 2014. (/jspui/handle/123456789/194).

62. Asifa Nahid, Haritha Pavani, Development and validation of HPLC method for simultaneous determination of Ranolazine and Dronedarone in bulk and pharmaceutical dosage forms. Indian Journal of Research in Pharmacy and Biotechnology. 2(6):1524-1528.

63. Sharma A, Prakash D, Singh SK. Development and validation of UV spectrophotometric method for the estimation of ranolazine in bulk drug and pharmaceutical formulation. Int J Chem Tech Res. 2010 Oct; 2(4):1945-8.

64. Kmail N, Shantier S, Gadkariem E. Derivative Spectrophotometric Methods for the Analysis and Stability Studies of Ranolazine in Bulk and Dosage Forms. Hacettepe University Journal of the Faculty of Pharmacy. 2022; 42(1):13-21.

65. Killedar S, Pawar A, Nadaf S, Nale A, Tamboli U, Pishawikar S. Novel analytical method development for some amide group containing drugs using Bougainvillea spectabilis bract extracts. Asian Pacific Journal of Tropical Medicine. 2014 Sep 1; 7: S560-7.

66. Vyas AJ, Raval DH, Patel AI, Patel AB, Patel NK, Chudasama A. First order derivative spectrophotometric estimation of ranolazine and metformin hydrochloride in synthetic mixture. Chem Methodol. 2020; 4: 605-13.

67. Patel J, Patel P, Mashru R. Novel UV-spectrophotometric and RP-HPLC method development and validation of simultaneous estimation of ranolazine and metformin HCl: A statistical analysis. Pharm Innov J. 2020; 9: 50-7.

68. Vyas K, Soniwala M, Vyas A, Shah N. Development of a UV visible spectrophotometric method for simultaneous estimation of ranolazine and metoprolol. Journal of Drug Delivery and Therapeutics. 2022 May 2; 12(3-S): 64-72.

69. Nahid A, Pavani H. Development and validation of HPLC method for simultaneous determination of Ranolazine and Dronedarone in bulk and pharmaceutical dosage forms. Indian Journal of Research in Pharmacy and Biotechnology. 2014 Nov 1; 2(6): 1524.

Received on 05.08.2025 Revised on 24.08.2025

Accepted on 14.09.2025 Published on 20.09.2025

Available online from September 30, 2025

Research J. Engineering and Tech. 2025; 16(3):127-136.

DOI: 10.52711/2321-581X.2025.00012

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