INTRODUCTION:

Topical forms of Vitamin A have been widely used since 1971. Topical retinoids are either natural or synthetic. Adapalene, tazarotene, and bexarotene are synthetic retinoids and have different chemical structures, making it more challenging for the metabolic pathway. Tazarotene is an acetylenic synthetic retinoid introduced in 1997 and has a different chemical structure than other retinoids. It does not occur naturally and is approved by the United States of Food and Drug Administration (US FDA) for acne vulgaris, psoriasis, and photodamaged.¹

Topical Therapies for Psoriasis:

treatment. By penetrating the stratum corneum interstices, emollients enable some of the damage to heal² reversing part of the inflammatory reaction to this injury. In addition to improving the absorption of topical corticosteroids or coal tar, keratolytic treatments like salicylic acid ointment soften and facilitate the elimination of psoriatic plaques ³. Other topical therapies are more successful than emollients and keratolytic agents, which have extremely limited efficacy. Regardless, topical therapy alone isonly suited for individuals with mild-to-moderatepsoriasis, and of these only a tiny fraction of patients achieve a high level of response (defined as 76- 100% improvement) that is maintained for even a fewmonths.⁴ Adverse effects and cosmetic issues are additional constraints on all topical treatments.

Retinoids can affect gene transcription both directly and indirectly for example by acting directly on genes that contain retinoid response elements, or indirectly by inhibiting transcription. They induce their effects mainly through binding to and activation of two families of nuclear retinoid receptors: retinoic acid receptor (RAR) and retinoid X receptor (RXR)⁵. Each family has 3 receptor subtypes: α, β, and γ. Receptor selective retinoids have been developed with the aim of maximal efficacy with minimal adverse effects. Tazarotene is a synthetic receptor selective retinoid, whose active metabolite, tazarotenic acid, binds selectively to RAR β and γ (Chandraratna 1996)⁶.

Important of Tazarotene:

Tazarotene is important for its effectiveness in treating inflammatory and non-inflammatory skin conditions, such as plaque psoriasis, acne, and signs of sun-damaged skin. It works by slowing down rapid skin cell growth, reducing inflammation, and promoting healthier skin cell turnover, which can lead to a reduction in thick scales, acne lesions, and signs of aging like fine wrinkles and discoloration.

Prescription required: Tazarotene is a prescription medication.

Potential side effects: Common side effects include skin dryness, redness, irritation, peeling, and a burning or stinging sensation.

Pregnancy: Tazarotene is a teratogen and should be avoided by women who are pregnant or contemplating pregnancy, and an effective form of birth control is necessary for women of child-bearing potential.

Analytical Method in Medicine:

Tazarotene (TZR) is the first topical receptor-selective retinoid prodrug derived from vitamin A used for management of plaque psoriasis and efficacious in dealing of acne vulgaris, and photo aging. As per US food and drug administration (FDA), 0.1% strength of TZR is permitted for the treatment of acne. This article draws attention to various advanced and conventional analytical methods. The hyphenated and conventional chromatographic techniques such as LC-MS/MS and HPTLC, HPLC respectively. Moreover, spectrophotometric methods like UV/visible spectroscopy are also used to quantify TZR as an active pharmaceutical ingredient and its formulations, especially in topical preparations. Moreover, the TZR is required for alternative methods for routine quality control and to estimate TZR in pharmaceutical dosage form especially in pharmacokinetic studies of topical preparation. This write up focus on critical review of characteristics, uses and the information about the physicochemical, pharmacokinetics properties, mechanisms of action and more emphasis on different analytical methods for estimation of TZR in pharmaceutical formulations.⁷

ANALYTICAL TECHNIQUES:

Analytical techniques for tazarotene include chromatography (HPLC, RP-HPLC, HPTLC, TLC, Paper Chromatography, Gas Chromatography). which separates and quantifies the drug, and spectroscopy (UV-Vis, IR, NMR, MS) derivative and derivative ratio spectrophotometry), which measures its concentration in solutions.

1] HPLC Instrumentation and Chromatographic Condition:

The LC system of Waters Alliance HPLC with PDA detector was used for this study and chromatographic separation was achieved on Waters Symmetry C18 (150 mm X 3.9mm, 5µm) column as stationary phase with binary gradient mode.

The gradient program was used as time (min)/mobile phase-A (%)/mobile phase-B (%); 0.0/100/00, 10/100/00, 15/75/25, 45/75/25, 47/100/00, 55/100/00. Where, mobile phase-A was Buffer: organic modifier, 40:60 v/v (v/v) and mobile phase-B, Organic modifier. Buffer was 10mM potassium dihydrogen phosphate, pH adjusted to 3.0 with orthophosphoric acid and filtered through 0.22µm membrane and organic modifier was methanol: tetrahydrofuran, 95:5 v/v. Column oven temperature was maintained at 30°C and injection volume for each study preparation was 50 µL. Eluent with flow rate of 1.0 mL/min was monitored at 325 nm with PDA detector. Acetonitrile: water, 80:20 v/v was used as a final diluent. The stress degraded samples and the solution stability samples were analyzed using a PDA detector covering the range of 200-400nm.

Diluted Standard Solution Preparation:

Standard solution was prepared by dissolving standard substance in diluent to obtain solution containing 1 µg/mL of TAZ.

Sample Solution Preparation:

An accurately weighed sample equivalent to 5mg of TAZ was taken into 50mL volumetric flask. About 35 mL of diluent was added to this volumetric flask and sonicated in an ultrasonic bath for 15min with intermittent shaking, diluted to the volume with diluent, mixed well, centrifuged at 7500rpm for 15min. Supernatant solution was filtered through 0.2µm PVDF syringe filter.

Mobile Phase and Gradient Optimization:

A method development was started with Waters symmetry C18 150mm x 3.9mm, 5µm column as stationary phase. Mobile phase A was buffer (10mM KH2PO4 with pH 3.0 by orthophospheric acid), while mobile phase B was methanol: tetra hydrofuran, 95:5 v/v. For gradient optimization first programme was time (minutes)/ mobile phase B(%); 0/60, 10/60, 15/80, 25/80, 30/60, 35/60 and obtained chromatogram in order to achieve better chromatographic separation (between impurity-A and unknown impurity), mobile phase-B was modified as time (minutes)/mobile phase B(%); 0/60, 10/60, 15/70, 45/70, 47/60, 55/60.

Fianally, in order to increase the mobile phase stability (to prevent microbial growth) organic component was added to buffer of mobile phase A, in the ratio of buffer: organic modifier, 40:60 v/v.

Diluent Selection:

Initially, methanol: water, 80:20 was used as diluent with consideration of solubility of TAZ and related impurites in methanol for better extraction and water was incorporated to reduce the rate of evaporation. But, during the forced degradation study (base hydrolysis), significant level of TAZ impurity-A was observed.

Analytical Method Validation:

After satisfactory development of method it was subjected to method validation as per ICH guideline^8,9. The method was validated to demonstrate that it is suitable for its intended purpose by the standard procedure. Analytical method validation was carried out by means of system suitability, accuracy, precision, linearity, robustness, solution stability and filter compatibility.

System Suitability:

The system suitability was performed by diluted standard solution and impurities spiked solution (1 µg/mL Imp-A, Imp-B, Imp-C and 100µg/mL of TAZ). The system suitability parameters were evaluated and found to be within the limits.

Linearity:

Linearity was performed for imp-A, Imp-B, Imp-C and TAZ. The correlation coefficient value was found to be more than 0.999. Bias was less then 2.0% for each case.

Method Precision:

The method was found to be precise with six sample preparations by spiking the impurities at 0.3% level. The % RSD of Imp-A, B, C in six sample preparation was found to be less than 6.0%

Accuracy:

Accuracy was established by recovery methodology by spiking each of imp-A, B, C, at six different levels, in triplicate preparations, starting from LOQ to 150% of impurity specification of drug product The recovery results for all impurities were found to be between 90-110%.

Solution Stability:

The solution stability of the standard and spiked sample preparation in diluent was studied for 72hrs. at bench top. The solution under study was compared with freshly prepared standard solution. The samples solution stability was established upto 72hrs and found satisfactory.

Robustness:

The robustnes was demonstrated by varying; flow rate, mobile phase buffer pH and column oven temperature. The method was found to be robust with respect to flow rate, pH and column temperature without any significant changes in system suitability parameters and relative retention time of impurities in sample.

RP-HPLC Method:

A stability-indicating RP-HPLC method has been developed and validated for the determination of tazarotene (TA) in topical pharmaceutical dosage formulation.

The optimization of column selection and mobile phase selection were done simultaneously. An isocratic method was employed using a buffer (0.02M ammonium acetate pH 2.5 with glacial acetic acid), acetonitrile, and methanol in the ratio of 50:25:25 v/v/v, respectively, as the mobile phase. The X-Terra™ C18 (50×4.6mm, 5μ) column with a flow rate 1.5mL/min at column temperature 40°C was used in the HPLC, equipped with a photodiode array detector. TA peak fronting was observed and the peak was eluted too late. To reduce the run time and improve the TA peak shape, an attempt was made by replacing methanol with acetonitrile from the mobile phase component which then became 0.02M ammonium acetate (pH 2.5 with glacial acetic acid) and acetonitrile in the ratio of 50:50 v/v. The column was changed to the Waters X-Bridge™ C18 (50×4.6mm, 3.5μ) for better peak shape. The TA peak eluted at 8.0 minutes but the PE and MP peaks were co-eluting in same retention time in the column void. To separate PE from the MP peak, an attempt was made with gradient elution with the mobile phase (0.02M ammonium acetate pH 2.5 adjusted with glacial acetic acid) as solvent-A and acetonitrile as solvent-B. PE was separated from the MP peak, and also the peak tailing 1.0 was observed for TA. The peak shapes for all of the components were good, but blank interference was observed at the retention time of MF. To remove the blank interference at the retention time of MF, solvent-A was changed to a 0.1%v/v orthophosphoric acid buffer, while keeping acetonitrile as solvent-B with the same gradient mode. As a result, no blank interference was observed. But when the base degradant sample was injected, the MF peak was eluted along with the base degradant peak. To separate the MF peak from the base degradant peak, the gradient programme was modified as time (min)/mobile phase-A (%)/mobile phase-B (%); 0.0/90/10, 1.5/90/10, 4.5/78/22, 8/50/50, 10.5/50/50, 15/5/95, 17/90/10, 20/90/10. While the flow rate was 1.5mL/min and the column temperature was 50°C, the MF peak was separated from the base degradant peak. Good peak shape for all of the components with well-resolved degradant peaks were observed. Also, the resolution between the PE and MP peak was greater than 2.7. The wavelength was selected by injecting a known concentration of TA into the HPLC with a PDA detector, and was evaluated for the UV spectra of each component. A common wavelength for the simultaneous determination of all the components was selected as 256 nm by overlaying the spectra and wavelength at which all components had significant absorbance.

After satisfactory development of the method, it was subjected to method validation as per ICH guidelines [10,11]. The method was validated to demonstrate that it is suitable for its intended purpose by the standard procedure to evaluate adequate validation characteristics (system suitability, accuracy, precision, linearity, limit of detection, limit of quantification, robustness, solution stability, filter compatibility, and stability-indicating capability).

Chromatographic Conditions:

All chromatographic experiments were performed using the Waters X-Bridge™ C18 (50×4.6mm, 3.5μ) column. The optimized mobile phase consisted of 0.1%v/v orthophosphoric acid in water as solvent-A and acetonitrile as solvent-B. Solvents-A and -B were filtered through a 0.22μm nylon membrane filter and degassed under vacuum prior to use. The separation of PE, MP, PP, MF, TA, and all impurities was achieved by gradient elution using solvent-A and solvent-B. A mixture of acetonitrile and water in the ratio of 80:20 (v/v), respectively, was used as diluent. A gradient program was used as time (min)/mobile phase-A (%)/mobile phase-B (%); 0.0/90/10, 1.5/90/10, 4.5/78/22, 8/50/50, 10.5/50/50, 15/5/95, 17/90/10, 20/90/10, at a flow rate 1.5 mL/min at 50°C, detection wavelength 256 nm.

Standard Solution Preparation:

The stock solutions of PE (400μg/mL), MP (400 μg/mL), PP (400μg/mL), MF (200μg/mL), and TA (200 μg/mL) were prepared by dissolving an appropriate amount of standard substances in diluent, separately. Working standard solution was prepared by mixing the above stock solutions of PE, MP, PP, MF, and TA with a final concentration of 40μg/mL, 40μg/mL, 4μg/mL, 20 μg/mL, and 20μg/mL, respectively.

Sample Solution Preparation:

An accurately weighed 1g sample (equivalent to 1mg of TA, 1mg of MF) was taken into a 50mL volumetric flask. About 35mL of the mixture of acetonitrile and water (80:20, %v/v) was added to this volumetric flask and sonicated in an ultrasonic bath for 15 min with intermittent shaking, diluted to the volume with a mixture of acetonitrile and water (80:20, %v/v), and mixed well. A portion of the solution was filtered through a 0.22μm nylon syringe filter and the filtrate was collected after discarding the first few milliliters.

Paper Chromatography:

Paper chromatography is generally not used for the analytical quantification or quality control of tazarotene in pharmaceutical applications. Modern analytical reviews and methods focus on more advanced and precise techniques like High-Performance Thin-Layer Chromatography (HPTLC), High-Performance Liquid Chromatography (HPLC), and hyphenated techniques such as LC-MS/MS.

Why Paper Chromatography Is Not Used:

Paper chromatography is an older, low-cost, and simple qualitative technique primarily based on partition chromatography. It has limitations for complex, non-volatile, or trace-level analysis, and generally provides less accurate and less reproducible results compared to modern instrumental techniques like HPTLC or HPLC. Reviews of analytical methods for tazarotene do not list paper chromatography as a standard or validated technique for its analysis.

GAS Chromatography:

A highly sensitive gas chromatography-mass spectrometry quantitative method was developed for 3,3-dimethylallyl bromide genotoxic impurity in the tazarotene drug substance. The method was validated by following the International conference for harmonization Q2 (R1) guideline for the limit test of 3,3-dimethylallyl bromide genotoxic impurity in the tazarotene drug substance. The 148m/z, 150m/z, and 69m/z ions were selected to get a response in mass spectrometry for 3,3-dimethylallyl bromide. DB-1 (30mm × 0.25 × 1.0µm) column was used for the separation of 3,3-dimethylallyl bromide from the sample matrix. The limit of detection and limit of quantitation for 3,3-dimethylallyl bromide in this method was 0.7 and 2.1ppm, respectively. The method was precise and accurate at limit of quantitation level, 1.7% RSD was observed at limit of quantitation precision and 100.9% recovery observed at limit of quantitation accuracy, it was linear from the 5 to 20 µg/L, the correlation coefficient (r) was 0.9996.¹²

Thin Layer Chromatography TLC:

Thin layer chromatography (TLC) analytical techniques for tazarotene use a silica gel stationary phase and a mobile phase like toluene-methanol (\(9.0:1.0,v/v\)). Tazarotene is quantified using a densitometric scanner at 327 nm, with a validated method providing good precision, accuracy, sensitivity, and robustness for the separation and analysis of tazarotene in pharmaceutical preparations.

Pure sample:

Standard (MF), (MIC) and (GM) were kindly donated by SIGMA Pharma Co., Quesna, Egypt. Their purity was found to be 100.08±0.39², 100.17±0.13³⁶, and 100.02±0.30⁹ according to reported methods, respectively.

Solvent:

Methanol and chloroform (Analar grade) and formic acid solution was supplied from (Adwic, El Nasr pharmaceutical.

Standard Solutions Stock Solutions:

Solutions were prepared in methanol of concentrations: 1mgm L^-1 MF, 2mgm L^-1 MIC mL^-1 and in methanol and water of concentration 4 mgmL^-1 GM.

Working solutions: Working solutions were freshly prepared by further dilution of suitable volumes from each stock solutions with me 200μgmL-1solutionsthanol to get solutions of final concentration for TLC spectrodensitometric method, 0.5mgmL^-1 MF, 1mgmL^-1 MIC, and 2mgmL^-1 GM; for chemometric method, 100μgmL^-1 MF, from 200 to 400μgmL MIC, and 2000μgmL^-1.

TLC:

To improve topical delivery and to reduce adverse effects, certain locally applied drugs need to contain various lipophilic excipients in their formulations that are insoluble in organic solvents commonly used in reversed-phase liquid chromatography (RPLC). TLC/HPTLC-based methods are considered a good alternative to RPLC in this case for the routine analysis of these pharmaceutical products. For TLC/HPTLC, there are no limitations for the choice of mobile phase components, and drugs and lipophilic excipients can be dissolved in solvents that will later evaporate when samples are applied to the layer. A further benefit is that an extraction procedure is no longer required and chromatogram can be developed without interference between the drug and excipients.

Spectroscopy:

spectrophotometric methods like UV/visible, IR, NMR, MS spectroscopy also used to quantify TZR as active pharmaceutical ingredient and its formulations, especially in topical preparations. Moreover, the TZR is required alternative methods for routine quality control and to estimate TZR in pharmaceutical dosage form especially in pharmacokinetic studies of topical preparation.

UV-Visble Spectroscopy:

Chemical structure of Tazarotene:

Structure of Tazarotene

The Tazarotene (Molecular Formula- C21H21NO2S) is third generation topical retinoid prescribed in the form of topical gel, cream, lotion etc. It is a specific retinoid having efficacy in the topical treatment of psoriasis.¹³

UV spectrophotometric methods have been reported for the simultaneous estimation of various drugs in various pharmaceutical products. The spectrophotometric method is not reported for Tazarotene. In our research work, we developed a method has been made for these drugs.¹⁴

MATERIAL AND METHODS:

The double beam UV-Visible spectrophotometer is used in our research work. Tazarotene was purchased from Sigma-Aldrich. Methanol was used as the main solvent system.

Analytical Wavelengths Determination:

The required dilutions were prepared for both drug Tazarotene with the prepared solution (standard), in spectrum mode scanned from the range 200 - 400nm.

Preparation of Stock Solution:

Tazarotene Standard Stock Solution:

Tazarotene (100mg) was accurately weighed and then transferred the sample of drug to a 100mL volumetric flask. Now dissolved the sample with methanol, the flask was shaken and final volume was made using methanol as solvent. Ten milliliters of solution was pipetted out from the prepared solution, transferred to 100mL volumetric flask, made the 100mL volumes with solvent (methanol). The concentration of prepared stock solution was 100μg/mL.¹⁵

Preparation of Different Analytical Concentrations:

For Tazarotene, from standard stock solution (100 μg/mL), pipetted out different concentrations 0.2, 0.4, 0.6, 0.8, 1.0mL and transferred to 10mL of separate volumetric flasks and the volume was made up to the mark with solvent (methanol). These concentrations were of 2, 4, 6, 8, 10μg/mL, respectively.

Method Validation:

Preparation of Calibration Curve:

For the calibration curve, take both the drugs in volumetric flask with same quantity (10mg) in different solvents (methanol, buffer saline solution pH 6.8), and the volume was made up to the mark. From the stock solution, different samples with concentration range from 2-10μg/mL were prepared by dilution with solvent (methanol). Spectrophotometrically measure the absorbance of each dilution at the respective wavelengths. The obtained data were processed using MS Excel computer program.^7,10,11,12

Linearity:

Calibration plots were prepared in various solutions and data were analyzed by applying linear regression analysis.

Accuracy:

Three known concentrations were made in the different solvent system (Phosphate buffer saline 7.4: Ethanol 1:1 v/v, Methanol and Methanol: Chloroform 1:1 v/v) and their absorbance values were measured at the respective λmax concentration values were calculated from absorbance value and compared with the actual used concentration.

Precision:

Drug solution of a known concentration was prepared in solvent and scanned on both drugs absorbance maxima, for Halobetasol 238 and Tazarotene 352 respectively. The process of precision was for the three days.

Repeatability:

Both drugs are analyzed on both wavelengths. The scanning numbers are six times of same concentration at the respective wavelength. After that, the mean absorbance of six scanning was calculated and %RSD with their standard deviations.

Limit of Detection and Limit of Quantification (LOD and LOQ):

The limit of detection is the method for smallest analytical concentration in drug samples that can be reliable to zero distinguish whereas limit of quantification is smallest analytical concentration that can determine with repeatability. The calculations of LOD and LOQ.

IR Spectroscopy:

Fourier-Transform Infrared (FTIR) spectroscopy is commonly used for identifying and characterizing tazarotene, confirming its structure, assessing purity, and analyzing interactions with excipients in formulations. Review articles often mention FTIR as one of several analytical techniques without detailing fine-grained methodology.

NMR Spectroscopy:

Review articles indicate that Nuclear Magnetic Resonance (NMR) spectroscopy is a key method for identifying and characterizing the structure of tazarotene, its degradation products, and impurities. While techniques like HPLC and LC-MS are used for routine quantification, NMR is essential for structural confirmation of the active pharmaceutical ingredient and related substances, aiding in the understanding of degradation pathways and pharmaceutical quality assessment.

Since 1H NMR is sensitive to compounds bearing protons, it can be regarded as a quasi-universal detector, especially for low molecular weight organic molecules. Protons are detected with the same sensitivity, regardless of their chemical environment; thus, the requirement of determining response factors is avoided.

Therefore, NMR ranks among the most informative methods, being a key analytical tool for identification, authentication, detailed structural analysis and elucidation of organic compounds (including stereochemistry and even dynamic effects), and quantitation of unknown natural and synthetic compounds in their mixtures in a single run, provided their signals are well separated¹⁶.

LATEST RESEARCH DEVELOPMENT:

Research on tazarotene continues to expand our understanding of this medication and explore new ways it might help patients. Recent studies have focused on improving how we use tazarotene and finding new applications for this versatile medication.

One exciting area of research involves new formulations designed to reduce irritation while maintaining effectiveness. Researchers have developed lower-concentration options (0.045%) that may provide benefits similar to higher concentrations but with fewer side effects. These advances may make tazarotene more comfortable for people with sensitive skin.

Scientists are also studying combined formulations that pair tazarotene with other medications in a single product. These combinations aim to provide multiple benefits while simplifying treatment routines. Research shows that certain combinations may work better than either medication alone while potentially reducing side effects.

For acne treatment, recent studies have examined how tazarotene compares to newer treatments and how it might complement other acne therapies. This research helps doctors make better recommendations about when to use tazarotene and how to combine it with other treatments for optimal results.

In psoriasis research, scientists are investigating how tazarotene affects specific inflammatory pathways involved in the disease. This deeper understanding may lead to more targeted treatments or help identify which patients will respond best to tazarotene therapy.

Researchers are also exploring tazarotene's potential for treating additional skin conditions beyond its current approved uses. Early studies suggest it might help with certain types of skin discoloration, scars, and other disorders involving abnormal skin cell growth.

As research continues, we expect to gain more insights into how tazarotene works and develop better strategies for using it effectively while minimizing side effects.

CONCLUSION:

Available evidence indicates that in the short term at least topical tazarotene is safe and effective for the treatment of photodamaged skin. Furthermore, the clinical improvement seen in the tazarotene treated groups is consistent between the different trials. Adverse effects associated with tazarotene are mainly mild to moderate cutaneous irritation, which in clinical practice could perhaps be avoided by a more gradual institution of treatment.

The longest trial to date suggests that continued clinical improvement can occur beyond one year of treatment, and further trials to assess optimum duration of therapy and appropriate “maintenance” regimes to guide clinical practice are warranted.

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Received on 05.12.2025 Revised on 12.01.2026

Accepted on 07.02.2026 Published on 02.07.2026

Available online from July 15, 2026

Asian J. Res. Pharm. Sci. 2026; 16(3):271-277.

DOI: 10.52711/2231-5659.2026.00040

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