A review on HPLC Methods for Estimation of Travoprost  in Combined and Single Pharmaceutical formulation and Bulk

V.Swetha*, V.Sahithi, B.Anusha

Department of pharmaceutical analysis, School of pharmaceutical sciences and Technoligies

JNTUK, Kakinada, India 433003

*Correspondence: Swethatarunya.11@gmail.com , Mobile no: +91 9949178567

DOI: https://doi.org/10.71431/IJRPAS.2025.4610        

INTRODUCTION

Increased intraocular pressure (IOP) is a major risk factor for glaucoma, a condition that ranks as the second leading cause of blindness across the globe.¹ High IOP levels may directly harm ganglion cell axons and decrease blood supply to the optic nerve, leading to ischemia and neurodegeneration, even if the exact pathophysiology of glaucoma is yet unknown. The sole known treatment for glaucoma is lowering intraocular pressure (IOP).^2,3 Prostaglandin analogues (PGs) are a relatively new family of anti-glaucoma medications that function by lowering eye pressure through prostaglandin FP receptors found in the ciliary muscle and trabecular meshwork.⁴

Physiochemical properties

Travoprost, is chemically, propan-2-yl(5Z)-7-[(1R,2R,3R,5S)-3,5-dihydroxy-2-[(1E,3R)-3-hydroxy-4-[3-(trifluoromethyl)phenoxy]but-1-en-1-yl]cyclopentyl]hept-5-enoate, differentiate itself from is a synthetic isopropyl ester prodrug of a prostaglandin F2alpha (F2α) analogue and selective FP prostanoid receptor agonist.⁶ The molecular weight and molecular formula of Travoprost is 500.5477 and C₂₆H₃₅F₃O₆ respectively.⁶

Fig.1 Chemical structure of Travoprost

Mechanism of Action

Travoprost is a pro-drug.  After injection, travoprost is absorbed through the cornea and hydrolysed to its active metabolite, travoprost free acid.  The ester moiety of the free acid facilitates penetration into the aqueous humour. While the specific mechanism of travoprost is uncertain, it is thought to be linked to its complete agonist action at the prostaglandin FP receptor. ⁷ Travoprost free acid lowers intraocular pressure by attaching to the FP receptor, which increases the drainage of aqueous humour through both the trabecular meshwork and uveoscleral routes.

Pharmacokinetic and Pharmacodynamic (PK and PD)

Travoprost shows a strong and selective binding to the prostaglandin FP receptor, acting as a full agonist at very low (nanomolar) concentrations. It does not significantly interact with other prostaglandin or non-prostaglandin receptors.

The primary aim of this study is to outline HPLC methods for determining the amount of Travoprost in different formulations and sample types, either on its own or in combination with other substances.

. The compiled data may be explored for the studies on analysis of Travoprost. The prominent hplc methods with Ultraviolet-Visible and PDA Detector for the study of travoprost.

High-Performance Liquid-Chromatography (HPLC)

HPLC is a type of column chromatography that pumps a sample solution in a mobile phase at high pressure through a column containing stationary phase. It is an instrumental analytical technique that can resolve, identify, and quantify elements present in a mixture.⁹ HPLC's capacity to detect samples at trace concentrations as low as parts per trillion has led to a number of industrial and scientific uses, including the analysis of chemicals, medicinal compounds, environmental samples, and forensic investigations. The HPLC method for determining travoprost from bulk material and pharmaceutical dosage forms in combination or single analysis was developed with the help of several researchers.¹⁰

We have attempted to provide a summary of the many chromatographic conditions used to construct HPLC procedures, their results, and a conclusion in this review paper. Out of all the HPLC techniques examined, it was discovered that the C18 column was the only one used to separate travoprost because it had the highest level of hydrophobicity and a sorbent phase that retained the ligand molecules.

Pharmaceutical methods of Travoprost

Pharmacopoeial Methods

Travoprost is the official drug in the United States Pharmacopeia (USP). USP have reported a HPLC procedure, USP describe the analysis of Travoprost using reversed-phase column 4.6-mm × 15-cm; packing L1 through the mobile phase parts consisting of Acetonitrile and 2.18 mg/mL of sodium 1-octanesulfonate in water. (0.5 mg per mL). Adjust with phosphoric acid to a pH of 3.5. (17:33), at flow rate of 2.0 ml/min and the detection of Travoprost at a wavelength of 220 nm.

Vendor (Merck) Column Characterization of Travoprost

Merck . examined Travoprost and Travoprost  free acid (Fluprostenol) in the pharmaceutical dosage form. Separation of these two drugs  was accomplished on a Chromolith High Resolution RP-18 endcapped, 100 x 4.6 m column by using acetonitrile, water and TFA as a solvent system with flow rate kept at 1.0 ml/min. Further, UV (Ultraviolet) detection of ISRA was monitored at 210 nm. The fraction assessment of the Travoprost elution as per the preparation reported to be in the range of guidliness proposed.

Fig.2 Travoprost Chromotogram

Table 1 :Vendor (Merck) Column Characterization of Travoprost

R. V. Valli Kumari et al. reported HPLC method to estimate the Travoprost and Timolol from Ophthalmic formulations. The linear relation between the peak area and concentration was recognized with a coefficient of correlation value of 0.9998. The developed method was set up precisely with Percentage recoveries for Travoprost and Timolol were 100.27 and 100.41% respectively, Travoprost and Timolol can be measured simultaneously in bulk and pharmaceutical dosage forms using a straightforward, accurate, and stable RP-HPLC method that has been designed and validated. The Intersil ODS (150X4.6mm, 5μ) column was used for the chromatographic separation, and the mobile phase was acetonitrile: buffer in a 10:90% v/v ratio. The effluent was measured at 213 nm, and the flow rate was 1 ml/min. Travoprost and Timolol were shown to have retention times of 2.516 and 6.198 minutes, respectively.

Fig.3 Travoprost Chromotogram of standard

For Timolol and Travoprost, the technique was linear over the concentration range of 12.5-75 μg/ml and 10-60 μg/ml, respectively. The repeatability analysis revealed that the procedure was exact, with a %RSD of less than 2. Travoprost and Timolol had respective percentage recoveries of 100.27 and 100.41%. The method's validity is demonstrated by the fact that all of the validation parameters were established and discovered to be within the bounds specified by ICH recommendations. Acid, base, peroxide, heat, UV, and neutral degradation stress conditions were applied to timolol and Travoprost. The degradation products demonstrated stability and the method's capacity to resolve them from the primary peak and associated contaminants.

FEI Luhua et all., To develop a technique for using HPLC to identify the related compounds of travoprost in travoprost and timolol maleate eye drops. Techniques Utilizing a gradient elution technique with 20 mmol·L-1 sodium heptane sulfonate solution (adjusted to pH 2.5 with phosphoric acid) as mobile phase A, methanol as mobile phase B, and acetonitrile as mobile phase C, the determination was carried out on a Phenomenex LUNA phenyl-hexyl column (3.0 mm×150 mm, 3 μm) with a detection wavelength of 220 nm. The column was heated to 40 °C, and the flow rate was 0.5ml/min, Travoprost and the contaminants were successfully separated under the ideal HPLC chromatography conditions. Travoprost, its related compound A, 15-epi diastereomer, 5-six-trans isomer, and 15-keto derivative calibration curves showed good linearities over the range of 0.02-219.29 μg·mL-1 (r=1.000 0), 0.02-3.10 μg·mL-1 (r=0.999 9), 0.02-3.87 μg·mL-1 (r=1.000 0), 0.02-4.37 μg·mL-1 (r=1.000 0), and 0.02-3.98 μg·mL-1 (r=0.999 9), respectively. The aforementioned chemicals have respective LODs of 2, 1, 2, 2, and 2 ng. Travoprost recovery rates were 99.7% on average (RSD=0.60%).

Jothieswari Dhamotharan et all.., Developed For the simultaneous quantitative assurance of Travoprost and Timolol maleate in an ocular arrangement, a simple, quick, and accurate RP-HPLC method was developed. Using a PDA finder and a PHENOMENOX C18, 15O×4.6 mm, 5 micron turn around stage logical segment, a chromatographic partition was achieved. The segment was passed through at a stream rate of 0.8 ml/min by the portable stage, which was made up of orthophosphoric corrosive (0.1%): methanol (60: 40 v/v). Using an isocratic method, the strategy was implemented at a frequency of 272 nm.

Performance attributes of HPLC method: (Table 2 ) ^(11-14)

Table 2: Performance attributes of HPLC method ^(11-14)

Acknowledgment:

I thankful to my guide for encouragement and supporting to finish this review.

CONCLUSION:

 The present review described the summary of hplc methods which are reported in the literature for the estimation of Travoprost not only in single dose but also in combined dosage form in bulk, pharmaceutical formulations as Travoprost, a prostaglandin analog, increases the outflow of aqueous humour, whereas timolol, a beta-blocker, decreases the formation of aqueous humour (fluid in the eye). They are frequently mixed together to create a single eye drop solution. The primary objective of the compilation of review is to collect maximum information available on hplc analytical methods of Travoprost and study it in detail. From this survey, it is revealed that very few analytical methods are obtainable on high performance liquid Chromotography techniques, in conclusion, pharmacokinetic research, quality control, and regulatory compliance for Travoprost all still depend heavily on HPLC. In pharmaceutical analysis, its ability to be tailored for routine and advanced analytical applications makes it a fine standard in pharmaceutical analysis.

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