Int J Curr Pharm Res, Vol 14, Issue 3, 17-21Review Article



Department of Pharmaceutical Chemistry, 1Department of Quality Assurance, S. S. R. College of Pharmacy, S. S. R. Campus, Sayli Road, Silvassa 396230, Dadra and Nagar Haveli, India

Received: 20 Jan 2022, Revised and Accepted: 22 Mar 2022


In this present situation increase in the number of diseases has been observed and several new medications are invented and have been developed to treat various disorders, which are approved by FDA. But before these drugs come to market it must undergo several procedures. The validation and analytical process of a new drug development helps in ensuring its purity and reliability. This process involves the use of various analytical techniques to collect data about the drug. This review includes various types of analytical techniques such as ultraviolet-visible spectrophotometric and some chromatography methods (High-performance thin-layer chromatography, High-performance liquid chromatography, gas chromatography), hyphenation techniques such as LC-MS of the newly approved drug in the year of 2021 have been discussed.

Keywords: Analytical method, FDA approved drugs, HPLC, UV-VIS, HPTLC, LC-MS


The development of an analytical method and validation are key parameters for discovering, developing and manufacturing of pharmaceutical substances. Method development could be a procedure of validating that an analytical method is appropriate to be used to live the concentration of an API in an exceedingly pharmaceutical dosage form as the amount of medicine that is launched into the market each year is rising. This drug can either be newly formulated or structurally modified of the main drug. Under these conditions, analytical procedures and standard methods of this latest medications might not be mentioned in the pharmacopoeias [1]. So, the development of analytical methods is necessary for such new approved drugs. Internal control laboratories use official test procedure for ensuring the identity, purity, and efficiency including the performance of drug products. To analyse the analyte there are several methods such UV Spectrophotometric, Ultra Performance liquid chromatography, High-performance thin-layer chromatography, High-performance liquid chromatography, Stability indicating High-performance liquid chromatography, LC-MS, Spectro-fluorimetry, GC-MS, etc. [2]. In the Pharmaceutical industry Qualitative and Quantitative determination of medicine, API, Raw materials, and biological samples, there are Analytical methods used like spectrometry and chromatography. These methods accustomed the identity, purity, potency and performance of medicine [3]. Analytical method development plays the most roles within the expansion in manufacture of pharmaceuticals.

The Food and Drug Administration (FDA) is a national government specialized agency of the United States. It was established in 1906 as the Department of Health and Human Services.


This organization mainly takes responsibility for governing and managing the safety of food, dietary supplements, therapeutic medications, vaccine, biological medicinal product and medical devices such as radiation-emitting devices, blood products, veterinary products and cosmetics.

Headquarters of FDA is situated in oak, Maryland. This organization controls 223 fields offices and Research Laboratories which are located across the 50different states.

In 2008, the FDA initiated for introducing its offices in other nations which include India, China, the United kingdom and Costa Rica Belgium.

Fig. 1: Flowchart of analytical techniques

UV spectrophotometric

The UV-Visible spectrometry is considered as most traditional instrumental system for analysis and it is finest methods used for determining analytes which are present in sample and in micro and semi micro amounts. It is concerned with detection of the effects of electromagnetic radiation interacting with absorbing entities such as atoms, molecules, or ions in the UV and/or visible spectrum [4]. It is useful for analyzing various substances such as biomolecular, inorganic and organic compounds. Results of this analytes are used for research purpose, industry, clinical trial laboratories, it is also helpful for environmental samples. Therefore, it is necessary to learn about the origin of the UV-VIS spectrum and its characteristics.


High-Performance Liquid chromatography (HPLC) is that leading distinctive separating instrument which is used for many aspects of drug manufacture and analysis. As HPLC is highly sensitivity and specific, it gives précised result. But, HPLC is widely used mainly for two reasons firstly Qualitative and analysis of unknown mixtures and secondly, Mixture’s separation for the later analysis. The separation mode depends totally on interacting relationship between the analyte, stationary Phase, mobile Phase [5].


In modern era, The High-performance thin-layer chromatography an evolved system of thin layer chromatography with superior separation functioning and detection limits than GC and HPLC. As it is an easily adaptable analytical method that may be used for either qualitative or quantitative applications. Separation takes place due to adsorption and partition, or each, depending on the form of adsorbents used on the plates and the development solvent system [6].


The Liquid chromatography-Mass spectrometry is considered as most potent analytical process which is sensitive and specific. The LC-MS is hyphenated analytical System which works in combination of two system, liquid-Chromatography(LC)and mass spectrometry (MS) [7]. Via passing throughout column the components of mixtures get separated in HPLC (LC), but this LC cannot differentiate the separated component so, Mass spectrometry helps to identify unknown substances and recognize and also useful to elucidate their structures. As, spectrum mixture is highly complicated because of overlapping spectra, Mass spectrometry individually isn’t helpful for identifying mixtures. Hence, they are used in combination for more accurate result. Therefore, it is possible to separate and determine relative atomic masses or molecule masses at the same time [8].

Table 1: Different analytical methods of newly approved drug in 2021

S. No. Brand name API Analytical method Description of the method Reference
1 Cabenuva Cabotegravir and rilpivirine RP-UPLC

System-Waters Acquity UPLC

Column-Azilent C18(150*4.6 mm)

Mobile Phase-orthophosphoric acid (60): acetonitrile (40)

Wavelength-245 nm

Flow rate-1 ml per min

Injection volume-10 ml


Total run time-6 min




System-HPLC system Waters Alliance LC (model 2695)

Column-Symmetry C18 (4.6 mm150 mm, 3.5µ)

Mobile Phase-0.1% HCOOH: ACN (80:20)

Wavelength-231 nm

Flow rate-1 ml/min

Injection volume-10 ml

Retention time-Cabotegravir-2.050 minRilpivirine-3.942 min

Linearity-Cabotegravir-20-300 g/ml Rilpivirine-30-450 g/ml

Total run time-5 min

2 Lupkynis Voclosporin LC/MS/MS

System-HPLC Applied Biosystems/MDS-Sciex API3000 Turbo Ion Spray (TIS)

Column-Zorbax SB C8,(2.1 mm12.5 mm)

Mobile Phase-0.02% (GAA) glacial acetic acid: 0.02 mmol sodium acetate

Injection volume-20 ml

LLOQ-1–200 ng/ml

Total run time-2 min

Mass spectrometric detection-Mode-positive ionization multiple reaction monitoring (MRM)m/z 1236.8→1112.8

3 Cosela Trilacicilib HPLC

System-e-2695 chromatographic system

Column-ODS Inertsil (150 mm4.6 mm, 3.5 µ)

Mobile Phase-(50)ACN: (50)

0.1% ortho phosphoric acid

Wavelength-220 nm

Flow rate-1 ml/min

Injection volume-10 ml

Retention time-4.358 min.

Linearity-3-45 µg/ml

Total run time-6 min

4 Fotivda Tivozanib RP-HPLC

System-Waters alliance liquid chromatography (model 2695)

Column-X~Bridge phenyl, (150 mm4.6 mm, 3.5 µ)

Mobile Phase-ACN: 0.1 % HCOOH (50/50)

Wavelength-216 nm

Flow rate-1 ml/min.

Retention time-4.07 min





System-Acquity UPLC, Nexera 2 series LC system connected (TQ-MS) triple quadrupole mass spectrometer API4000 (Sciex)

Column-BEH C18 (50 mm2.1 mm, 1.7 μm)

Mobile Phase-0.1% HCOOH in H2O: 0.1% HCOOH in ACN

Flow rate-0.3 ml/min

Injection volume-2 μl

Retention time-1.35 min

Linearity-0.5–5000 ng/ml

Total run time-15 min

Mass spectrometric detection-

m/z 455 to 341, m/z 456 to 341

5 Qelbree Viloxazine HPLC DAD


Column-Nucleosil RP-18 (250 mm 4.6 mm, 5 mm)

Mobile Phase-(pH 3.8) Buffer 20 mmol phosphate: ACN

Wavelength-220 nm

Flow rate-1 ml/min

Linearity-7-45 µg/ml

6 Nextstellis Drospirenone and estetrol  RP-UPLC

System-Waters Acquity UPLC system PDA detector

Column-Luna C18 (100 mm 2.6 mm,1.6 µ)

Mobile Phase-acetonitrile: 0.1% formic acid (70:30)

Wavelength-262 nm

Flow rate-1 ml/min

Injection volume-10 ml

Retention time-drospirenone-0.989 min

E4-1.878 min

Linearity-Drospirenone =3-45 µg/ml

E4 =14.2-213 µg/ml

Total run time-3 min

7 Lumakras Sotorasib RP-HPLC

System-Waters alliance liquid chromatography (model 2695)

Column-symmetry C18(150 mm4.6 mm, 3.5µ)

Mobile Phase-(70) Acetonitrile: 0.1% OPA (30)

Wavelength-221 nm

Flow rate-1 ml/min

Retention time-2.271 min






(Mouse plasma and Tissue homogenates)

System-Acquity UPLC® Shimadzu Nexera X2 (TQ/MS) triple quadrupole mass spectrometer (TIS)Turbo Ion Spray®

Column-BEH C18 (dp.1.7 µm, 30 mm2.1 mm) prefilter (2.1 mm, 0.2 µm)

Mobile phase-methanol: 0.1% HCOOH in water (50:50)

Flow rate-0.6 ml/min

Injection volume-2 ml

Retention time-0.5 min

Linearity-2–2,000 ng/ml

Mass spectrometric detection-positive (ESI) electrospray ionization (SRM)

Mode-selected reaction monitoring m/z 561.2 → 134.0



System-Shimadzu UFLC Prominence connected with Sciex 5500 triple quadrupole mass spectrometer

Column-Atlantis d C18 (5 µm,50 mm 4.6 mm)

Mobile Phase-0.2% HCOOH: ACN (25/75)

Flow rate-0.65 ml/min

Retention time-0.95 min

LLOQ-1.08 ng/ml

Total run time-2 min

Mass spectrometric detection-

Mode-multiple reaction mode (MRM)

(ESI) positive electro sprayionization) m/z 561.1→134.1 and 566.5→98.2

8 Truseltiq Infigratinib LC MS/MS

System-Agilent (1200) (TQ)Triple Quadrupole (6410)

Column-Agilent ZORBAX SB-C8 (1.8 μm,30*2.1 mm)

Mobile Phase-(20%) 0.1 percent formic acid in water: (80%)ACN

Flow rate-0.2 ml/min

Injection volume-5 ml

Retention time-1.54 min

Linearity-05–500 ng/ml

Total run time-2 min

Mass spectrometric detection-positive ion source electrospray ionization (ESI) Mode-multiple reaction monitoring (MRM)m/z 561 → 339

9 Kerendia Finerenone HPLC-MS/MS

System-e Milli Q system

Column-Luna C18(20 *2 mm, 3 µm)

Mobile Phase-ACN: 0.01 mol/l ammonium acetate (80:20 v/v)

Flow rate-1 ml per min

Injection volume-15 ml

Retention time-3.8 min

LLOQ-0.100 µg/l-200 µg/l

Mass spectrometric detection-

MRM, multiple reaction monitoring; m/z 379.05/217.90

10 Fexinidazole Fexinidazole UPLC MS/MS

System-Acquity UPLC system

Column-C18(1.7 µm,2.1 mm50 mm)

Mobile Phase-0.1% HCOOH: ACN

Flow rate-0.6 ml/min.

Injection volume-5 ml

11 Exkivity Mobocertinib LC-MS/MS

Column-ODS-3 column (i. d; 5 μm,50 mm4.6 mm)

Mobile Phase-A Mixture of water with 0.1% HCOOH: methanol 

Flow rate-0.5 ml/min.

Linearity-1-1000 ng/ml

Mass spectrometric detection-

Mode-Selected reaction monitoring (SRM) Positive ion source electrospray ionization (ESI), m/z 587.01 → 71.88




System-e-2695 chromatographic

Column-X-bridge phenyl column (150 mm x 4.6 mm, 3.5 µ)

Mobile phase-Acetonitrile: 0.1% o-phosphoric acid (OPA) (60:40 v/v)

Flow rate-1 ml/minWavelength-224 nm

Injection volume-10 ml

Retention time-2.271 min

Linearity-6-90 µg/ml

Table 2: List of no reported analytical methods of newly approved drug in 2021

S. No. Brand name API Manufacturing company
1 Verquvo Vericiguat Merck and Co., Inc.
2 Tepmetko Tepotinib EMD Serono, Inc.
3 Ukoniq Umbralisib Rhizen Pharmaceuticals
4 Evkeeza evinacumab-dgnb Regeneron Pharmaceuticals, Inc
5 Amondys 45 Casimersen Sarepta Therapeutics, Inc
6 Nulibry Fosdenopterin BridgeBio Pharma, Inc
7 Pepaxto melphalan flufenamide Oncopeptides Inc 
8 Azstarys serdexmethylphenidate and dexmethylphenidate Corium, Inc.
9 Ponvory ponesimod Janssen Pharmaceuticals, Inc., 
10 Zegalogue dasiglucagon Beta Bionics, Inc.,
11 Jemperli dostarlimab-gxly Glaxo Smith Kline LLC. 
12 Zynlonta loncastuximab tesirine-lpyl Avid Bioservices, Inc.
13 Empaveli Pegcetacoplan Apellis Pharmaceuticals, Inc.
14 Rybrevant amivantamab-vmjw Janssen Biotech, Inc., 
15 Pylarify piflufolastat F18 Progenics Pharmaceuticals, Inc
16 Lybalvi olanzapine and samidorphan Alkermes, Inc.
17 Brexafemme ibrexafungerp  SCYNEXIS, Inc. 
18 Aduhelm aducanumab-avwa Biogen MA, Inc.,
19 Rylaze asparaginase erwinia chrysanthemi (recombinant)-rywn Rylaze, Jazz, Pharmaceuticals, Inc
20 Rezurock Belumosudil Surface Logix, Inc
21 Bylvay Odevixibat Albireo Pharma, Inc.
22 Saphnelo anifrolumab-fnia AstraZeneca
23 Nexviazyme avalglucosidase alfa-ngpt Genzyme Corporation,
24 Welireg Belzutifan Peloton Therapeutics
25 Korsuva Difelikefalin Cara Therapeutics, Inc.
26 Skytrofa lonapegsomatropin-tcgd Ascendis Pharma, Inc
27 Tivdak tisotumab vedotin-tftv Seagen Inc.
28 Qulipta Atogepant AbbVie Inc.,
29 Livmarli Maralixibat Mirum Pharmaceuticals, Inc
30 Tavneos Avacopan ChemoCentryx, Inc.,
31 Scemblix Asciminib Novartis
32 Besremi ropeginterferon alfa-2b-njft PharmaEssentia Corporation
33 Voxzogo Vosoritide BioMarin Pharmaceutical Inc.
34 Livtencity Maribavir Takeda Pharmaceuticals USA Inc
35 Cytalux Pafolacianine On Target Laboratories, Inc
36 Tezspire tezepelumab-ekko Amgen, Inc.
37 Vyvgart efgartigimod alfa-fcab argenx SE
38 Leqvio Inclisiran Novartis AG
39 Adbry tralokinumab-ldrm LEO Pharma Inc


The development of the new method and validation of analytical methods is an essential step for developing of any pharmaceutical products. This review represents that many drugs been approved in the year of 2021, for use in market and based on the literature review it can be concluded that table 1 and table 2 drugs individually or its combination with another drug spectroscopy and chromatography methods are accessible and other remaining newly approved drugs there has been no spectroscopy or other information available. Although, many validation parameters of particular drugs have already reported, it is concluded that various analytical techniques like spectrophotometric, HPTLC, HPLC, GC-MS and LC-MS can be further developed for these formulations with the variations. It is vast opportunity for the development of new methods for newly approved drugs, as there is no well-established technique for some newly approved drugs or their combination with other drugs.




All the authors have contributed equally.


The authors hereby declare that there is no any conflict of interest involved in this paper.


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