A Concise Review on Analytical Profile of Ambrisentan

Rahul D. Rathod¹, Vikas R. Patil²*, Kuldip R. Patil³, Pawan I. Naik², Sopan Nangare⁴

¹QA Chemist, ICPA Health Products LTD., Ankleshwar- 393002, Gujrat, India.

²Assistant Professor, Department of Pharmaceutical Chemistry, TSPM’s, Trimurti Institute of Pharmacy,

Paldhi (Bk) - 425103, Dist. Jalgaon (MS), India.

³Assistant Professor, Department of Pharmaceutics, TSPM’s, Trimurti Institute of Pharmacy,

Paldhi (Bk) - 425103, Dist. Jalgaon (MS), India.

⁴Assistant Professor, Department of Pharmaceutics, H. R. Patel Institute of Pharmaceutical Education and Research, Shirpur - 425405 Dist. Dhule (MS) India.

*Corresponding Author E-mail: rdrathod15196@gmail.com, vikaspatil259@gmail.com, kuldippatil012@gmail.com, snangareopan@gmail.com

ABSTRACT:

Ambrisentan (AMB) belongs to the class of an antihypertensive drug. Generally, it is used to treat pulmonary arterial hypertension. It is a non-peptide, highly selective endothelin-1 antagonist of type A receptor. Herein, endothelin is the peptide that connects blood vessels and raises blood pressure (BP). In this case, AMB decreases blood pressure by inhibiting the effects of endothelin-1. As per literature, AMB was approved by the United States of food and drug administration (USFDA) in 2007. From its inception, there is several formulations are available in the market. Overall, there is a huge need to collect the analytical data reported by researchers owing to the major importance of AMB. Therefore, in the present review article, we have enlisted different analytical methods such as Ultraviolet (UV) visible spectroscopy, high-performance liquid chromatography (HPLC), ultra-performance liquid chromatography (UPLC), and liquid-chromatography-mass spectroscopy methods (LC-MS) for both qualitative and quantitative analysis of AMB in pharmaceutical and biologicals. In the future, this review article will assist researchers regarding the development of a new analytic method for AMB.

KEYWORDS: Ambrisentan; analytical methods; HPLC, UPLC; pharmaceutical dosage form.

INTRODUCTION:

Ambrisentan (AMB) is an orally active and highly selective antagonist of the endothelin-1 type A receptor. Therefore, it is most commonly used to treat pulmonary arterial hypertension. Moreover, AMB is offering a low risk of drug-drug interactions as compared to the other drugs. In addition, a dose of AMB is once a day only.¹

It has been reported that the endothelin-1 peptide plays a key role in the pathobiology of pulmonary arterial hypertension.^2,3 As per literature, AMB is authorized in Europe, Canada, and the United States (US) to enhance exercise capacity and postpone clinical deterioration. It is also approved in the US for usage with tadalafil to lower the risk of disease progression, hospitalization, and increase exercise capacity.⁴ Mainly, AMB is chemically known as (2S)-2-(4,6-dimethylpyrimidin-2-yloxy)-3- methoxy-3,3- diphenylpropionic acid.⁵ Figure 1 depicts the chemical structure of AMB.

Figure 1: Chemical structure of AMB

Analytical Account of AMB:

The comprehensive literature survey revealed that numerous analytical methods have been developed for evaluating AMB in bulk and pharmaceutical formulations. Principally, it includes UV/vis-spectroscopy, HPLC, UPLC, and LC-MS/MS. Interestingly, published literature regarding analytical methods for AMB estimation explained the estimation of AMB in various dosage forms as a single constituent. In this subsection, we have discussed the details of the UV-Visible spectroscopic and HPLC-based methods for AMB. In addition, we have summarized the UPLC and LC-MS methods for AMB.

Figure 2. Different analytical methods implemented for the estimation of AMB in a bulk and pharmaceutical dosage form.

1. UV-Visible spectroscopy method for AMB:

From its inception, UV-Vis spectrophotometry is mainly applied in the analysis of active in a pharmaceutical dosage form. In addition, the use of UV-Vis spectrophotometry increased rapidly over the last few years. The major benefits of this method are lower processing time, less labor requirement, cost-effectiveness, etc. In addition, the precision of UV-based analytical methods is also excellent. Notably, The quantitative assessment of a material's reflection or transmission qualities as a function of wavelength is known as UV spectrophotometry.⁶ Yunoos et al. reported method development and validated UV spectrophotometric method for the estimation of AMB in bulks and marketed formulations. Herein, 0.1 N sodium hydroxide (NaOH) has been used as a solvent. Afterward, UV-Vis spectrophotometric determination has been performed at 263.5nm, which gives a concentration linearity range of 10μg/mL - 50μg/mL with a correlation coefficient value of 0.9993. Moreover, the limit of detection (LOD) and limit of quantification (LOQ) was found to be 0.522μg/mL and 1.944μg/mL, respectively.⁷

2. High-performance liquid chromatography (HPLC) methods for AMB:

It has been revealed that the HPLC provides excellent specificity for the analysis of drugs in formulations as well as bulk. In addition, sufficient precision can be attainable by using HPLC based analytical method. Despite plenty of merits of HPLC for estimation of analyte in samples, several disadvantages affect the application of HPLC in the analytical field. According to the published literature, excellent specificity, precision, and accuracy can only be achieved if extensive system compatibility testing is performed before the HPLC analysis. As a result, the high cost is required to obtain high specificity, precision, and accuracy.⁶ Figure 1 showed the HPLC methods for AMB estimation in single and combined dosage forms.

Table 1. Summary of HPLC methods for the determination of AMB in a single and combined dosage form

Sr. No	Drug	Stationary Phase	Mobile Phase	Flow rate	Method of Detection	Retention time (Rt)	LOD	LOQ	Ref.
1.	AMB	Agilent Zorbax C18 column (250 mm × 4.6 mm I.D., 5 μm particle size)	0.1M Potassium dihydrogen phosphate buffer (pH was adjusted to 4.4 with Orthophosphoric acid) and methanol (30:70 v/v)	1.0 mL/ minutes	PDA at 210 nm	3.315 minutes	0.244 μg/mL	0.812 μg/mL	8
2.	AMB	Phenomenex C18 reverse phase column (250 × 4.6mm, 5μm)	Ortho phosphoric acid (0.05 %v/v): Methanol (20:80 %v/v)	1.0 mL/ minutes	PDA at 262 nm	5.8 minutes	0.404 μg/mL	1.225 μg/mL	9
3.	AMB	Sun Fire C18 column (150 mm length x 4.6 mm ID with 3.5 mm particle)	10 mM Potassium dihydrogen phosphate as solvent A at a pH adjusted to 2.5 with ortho-phosphoric acid in the water, and a mixture of Acetonitrile: Methanol (95:5, v/v) as solvent B	1.0 mL/ minutes	PDA at 225 nm	15.34 minutes	***	***	10
4.	AMB	Kromasil C18 column (250 3 4.6 mm, 5 mm)	Acetonitrile - Ammonium formate (pH 3.0; 0.02 M)	1.0 mL/ minutes	PDA at 262 nm	~15.8 minutes	0.2 mg/mL	0.6 mg/mL	11
5.	AMB	WELCHROM C-18 reverse phase column (4.6 x 250 mm, 5 μm)	10mM Phosphate Buffer (pH 6.0): Acetonitrile (50:50, v/v)	1.2 mL/ minutes	UV at 226nm	2.9 minutes	1.3267 μg/mL	4.020 μg/mL	12
6.	AMB	Phenomenex Luna C18 (250 × 4.6 mm, 5 μm)	0.02 M Ammonium acetate buffer (pH = 4.2), and Acetonitrile in 52:48 (v/v)	1.0 mL/ minutes	PDA at 215 nm	7.55 minutes	0.1 μg/mL	0.35 μg/mL	13
7.	AMB	Phenomenex C18 (150 × 4.6 mm, 5 μm)	10mM Ammonium acetate: Acetonitrile (65:35% V/V)	1.0 mL/ minutes	UV at 262 nm	2.696 minutes	***	***	14
8.	AMB	Agilent XDB-C18 (150 × 4.5mm, 5μ)	10 mM Ammonium ethanoate (pH-5.2 adjusted with acetic acid): Acetonitrile	1.0 mL/ minutes	289 nm	10.53 minutes	***	***	15
9.	AMB	C18 inertsil column (250 mm × 4.6 mm, 5 μm)	Water: Acetonitrile (30:70)	1.2 mL/ minutes	PDA at 264 nm	***	1.967	5.961	16
10.	AMB and TADA	Hypersil GOLD C18 column (150 mm × 4.6 mm internal diameter, 5 μm particle size)	Methanol: Water: Acetonitrile 40:40:20 (v/v/v)	0.5 mL/ minutes	UV/Vis at 260 nm	2.80 and 7.10 minutes	0.22 and 0.98 μg/mL	0.67 and 2.96 μg/mL	17

*PDA - Photo Diode Array.

***Not provided

Table 2. Summary of UPLC methods for the determination of AMB in the single and combined dosage form

Sr. No	Drug	Matrix	Stationary Phase	Mobile Phase	IS	Linearity	Ref.
1	AMB	-	Waters® BEH C18 column (2.1 mm x 50 mm, 1.7 µm -Waters®)	Formic: Trimethylamine: Water (1:1:998), pH 3.2	***	***	27
2	AMB	Human plasma	EVO C18 Column (50 X 2.1 mm, 1.7 μm)	Dimethyl sulfoxide: Acetonitrile and Acetonitrile: Water (1:1, v/v)	[^2H10]-Ambrisentan	0.1–200 ng/mL (plasma) and 0.1–10 ng/mL (Plasma ultrafiltrate)	28
3	AMB, Macitentan, Sitaxentan	Human plasma	BEH C18 column (1.7 μm, 2.1 mm x 50 mm)	Gradient consisting of water (eluent A) and methanol (eluent B). Both eluent contained of 0.10% (22 mM) formic acid and 0.013% (1.7 mM) ammonium acetate.	Ambrisentan-d10	20.28 - 2028 μg/l, 4.052 - 405.2 μg/l and 205.4 – 10 270 μg/l	29

***Not provided

3. Ultra-Performance liquid chromatography (UPLC) method for AMB:

The introduction of UPLC offers faster analytical separation procedures without sacrificing the high-quality results.¹⁸ Many laboratory specialists assured that the UPLC can eventually replace all current HPLC methods. In addition, it is an advanced technology that combines the unique characteristics and outperforms HPLC in several aspects, including greater chromatographic resolution, more sensitive analysis, less time consumption, ^19-23 reduced solvent use, and fast analytical speed. ^24-26 In this section, we have summarized the list of UPLC methods for the determination of AMB in the single and combined dosage forms. Table 2 depicts the summary of UPLC methods for the determination of AMB.

4. Liquid-Chromatography-Mass Spectroscopy methods (LC-MS) for AMB:

In recent years, the combination of LC/MS has gained a lot of attention for the analysis of interest analytes in complex samples with improved performance. In brief, after a thorough examination, ^30-32 LC/MS interfaces are divided into two categories namely interfaces for indirect and direct input of column effluent. A mechanical mechanism is employed to transmit the column effluent to the MS vacuum at an indirect introduction interface. A classic example of an indirect introduction type of interface is the transportation system. In the case of the direct introduction system, the column effluent flows directly into the mass spectrometric vacuum system via a tube. Mainly, the most straightforward method of linking LC and MS appears to be the direct introduction.³³ In this section, we have discussed the LC-MS methods for the determination of AMB in a dosage form (Table 3).

Table 3. Summary of LC-MS methods for the determination of AMB in a dosage form

Sr. No	Drug	Matrix	Stationary Phase	Mobile Phase	IS	Linearity (ng/mL)	Ref.
1	AMB	Rat Plasma	RP C18 Column	Water : Methanol (50:50 v/v)	Armodafinil	1-2000 ng/mL	34
2	AMB	***	Agilent XDB C18 column (150 x 4.6 mm; 5 mm)	10 mM Ammonium Ethanoate (pH = 5.2) and Acetonitrile	***	0.07–0.25 mg /mL	35

***Not provided

Unfortunately, there are only two reports are available based on the LC-MS method. In the future, there is huge scope to use the LC-MS method for estimation of AMB in single and combined dosage forms as well as in bulk samples.

CONCLUSION:

Estimation of analytes in single and complex samples is still a challenging step in the analytical field. Nowadays, significant progress has been done by researchers regarding the method development for the estimation of drugs and biomolecules in pharmaceutical formulations. In this review article, we have summarized the comprehensive data of various analytical and bioanalytical methods developed for AMB alone and in combination during the period of the last 15 years. In addition, the method along with their details such as solvent, mobile phase, stationary phase, retention time, etc. have been summarized. Mainly, HPLC, UV spectroscopy, UPLC, and LC-MS have been suggested for the determination of AMB in samples. Out of this, UV spectroscopy demonstrates the limited application of estimation of AMB, whereas UPLC and LC-MS based only a few methods are reported by researchers. Notably, HPLC-based methods have been major utilized for the analysis of AMB in a single and combined dosage form. In the future, this data will be of great help to researchers working in the analysis and bio-analysis field of AMB and others active in pharmaceutical and biological formulations. In conclusion, this review will open a new window for more information on new methods and changes on account of analytical methods of AMB in different dosage forms, and clinical samples.

ACKNOWLEDGEMENTS:

Authors are thankful to TSPM’s, Trimurti Institute of Pharmacy, Paldhi (Bk)-425103, Jalgaon, Maharashtra, India for providing necessary library facilities.

CONFLICT OF INTEREST:

The authors declare that no conflict of interest.

AMB      - Ambrisentan

WHO     - World health organization

BP          - Blood pressure

USFDA  - United States of food and drug administration

PAH       - Pulmonary arterial hypertension

US           - United States

UV          - Ultraviolet

UPLC   - Ultra performance liquid chromatography

HPLC   - High-performance liquid chromatography

LC-MS  - Liquid chromatography-mass spectroscopy

NaOH    - Sodium hydroxide

LOD      - Limit of detection

LOQ      - Limit of quantification

TADA    - Tadalafil

PDA       - Photodiode array

IS           - Internal standard

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Received on 14.06.2021 Modified on 30.10.2021

Accepted on 24.01.2022   ©Asian Pharma Press All Right Reserved

Asian J. Pharm. Ana. 2022; 12(1):56-60.

DOI: 10.52711/2231-5675.2022.00011