International Journal of Current Pharmaceutical Research

ISSN- 0975-7066 Vol 14, Issue 3, 2022

Review Article
RECENT METHOD DEVELOPMENT BY ANALYTICAL TECHNIQUES OF NEW FDA APPROVED
DRUGS IN 2021

ISHVARCHANDRA PARMAR , YOGI A PATEL1

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
Email: ijparmar2266@gmail.com
Received: 20 Jan 2022, Revised and Accepted: 22 Mar 2022
ABSTRACT
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
© 2022 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/)
DOI: https://dx.doi.org/10.22159/ijcpr.2022v14i3.1975 Journal homepage: https://innovareacademics.in/journals/index.php/ijcpr

INTRODUCTION methods used like spectrometry and chromatography. These

methods accustomed the identity, purity, potency and performance
The development of an analytical method and validation are key of medicine [3]. Analytical method development plays the most roles
parameters for discovering, developing and manufacturing of within the expansion in manufacture of pharmaceuticals.
pharmaceutical substances. Method development could be a
procedure of validating that an analytical method is appropriate to The Food and Drug Administration (FDA) is a national government
be used to live the concentration of an API in an exceedingly specialized agency of the United States. It was established in 1906 as
pharmaceutical dosage form as the amount of medicine that is the Department of Health and Human Services.
launched into the market each year is rising. This drug can either be
newly formulated or structurally modified of the main drug. Under FDA
these conditions, analytical procedures and standard methods of this This organization mainly takes responsibility for governing and
latest medications might not be mentioned in the pharmacopoeias managing the safety of food, dietary supplements, therapeutic
[1]. So, the development of analytical methods is necessary for such medications, vaccine, biological medicinal product and medical
new approved drugs. Internal control laboratories use official test
devices such as radiation-emitting devices, blood products,
procedure for ensuring the identity, purity, and efficiency including
veterinary products and cosmetics.
the performance of drug products. To analyse the analyte there are
several methods such UV Spectrophotometric, Ultra Performance Headquarters of FDA is situated in oak, Maryland. This organization
liquid chromatography, High-performance thin-layer controls 223 fields offices and Research Laboratories which are
chromatography, High-performance liquid chromatography, located across the 50different states.
Stability indicating High-performance liquid chromatography, LC-
MS, Spectro-fluorimetry, GC-MS, etc. [2]. In the Pharmaceutical In 2008, the FDA initiated for introducing its offices in other nations
industry Qualitative and Quantitative determination of medicine, which include India, China, the United kingdom and Costa Rica
API, Raw materials, and biological samples, there are Analytical Belgium.

Fig. 1: Flowchart of analytical techniques

 I. Parmar & Y. A. Patel
Int J Curr Pharm Res, Vol 14, Issue 3, 17-21

UV spectrophotometric HPTLC
The UV-Visible spectrometry is considered as most traditional In modern era, The High-performance thin-layer chromatography an
instrumental system for analysis and it is finest methods used for evolved system of thin layer chromatography with superior separation
determining analytes which are present in sample and in micro and functioning and detection limits than GC and HPLC. As it is an easily
semi micro amounts. It is concerned with detection of the effects of adaptable analytical method that may be used for either qualitative or
electromagnetic radiation interacting with absorbing entities such as quantitative applications. Separation takes place due to adsorption
atoms, molecules, or ions in the UV and/or visible spectrum [4]. It is and partition, or each, depending on the form of adsorbents used on
useful for analyzing various substances such as biomolecular, the plates and the development solvent system [6].
inorganic and organic compounds. Results of this analytes are used LC/MS-MS
for research purpose, industry, clinical trial laboratories, it is also
helpful for environmental samples. Therefore, it is necessary to learn The Liquid chromatography-Mass spectrometry is considered as
about the origin of the UV-VIS spectrum and its characteristics. most potent analytical process which is sensitive and specific. The
LC-MS is hyphenated analytical System which works in combination
HPLC of two system, liquid-Chromatography(LC)and mass spectrometry
(MS) [7]. Via passing throughout column the components of
High-Performance Liquid chromatography (HPLC) is that leading mixtures get separated in HPLC (LC), but this LC cannot differentiate
distinctive separating instrument which is used for many aspects of the separated component so, Mass spectrometry helps to identify
drug manufacture and analysis. As HPLC is highly sensitivity and unknown substances and recognize and also useful to elucidate their
specific, it gives précised result. But, HPLC is widely used mainly for structures. As, spectrum mixture is highly complicated because of
two reasons firstly Qualitative and analysis of unknown mixtures overlapping spectra, Mass spectrometry individually isn’t helpful for
and secondly, Mixture’s separation for the later analysis. The identifying mixtures. Hence, they are used in combination for more
separation mode depends totally on interacting relationship accurate result. Therefore, it is possible to separate and determine
between the analyte, stationary Phase, mobile Phase [5]. 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 RP-UPLC System-Waters Acquity UPLC [9]
and rilpivirine 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
Linearity-25-150µg/ml
Total run time-6 min
HPLC-MS System-HPLC system Waters Alliance LC (model 2695) [10]

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 [11]
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 [12]
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) [13]
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
Linearity-1.34-20.1µg/ml
LC-MS System-Acquity UPLC, Nexera 2 series LC system connected [14]

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S. No. Brand name API Analytical method Description of the method Reference
(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 System-HPLC-DAD [15]
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 RP-UPLC System-Waters Acquity UPLC system PDA detector [16]
and estetrol 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) [17]
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
Linearity-10-150µg/ml
LC MS System-Acquity UPLC® Shimadzu Nexera X2 (TQ/MS) triple [18]
(Mouse plasma and quadrupole mass spectrometer (TIS)Turbo Ion Spray®
Tissue Column-BEH C18 (dp.1.7 µm, 30 mm2.1 mm) prefilter (2.1
homogenates) 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
LC/MS-MS System-Shimadzu UFLC Prominence connected with Sciex [19]
(Mice) 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) [20]
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 [21]
Column-Luna C18(20 *2 mm, 3 µm)

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S. No. Brand name API Analytical method Description of the method Reference
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 [22]
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) [23]
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
HPLC System-e-2695 chromatographic [24]
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

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CONCLUSION 2008;874(1-2):57-63. doi: 10.1016/j.jchromb.2008.08.023,

PMID 18815080.
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and LC-MS can be further developed for these formulations with the method with a broad linear dynamic range for the
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AUTHORS CONTRIBUTIONS
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All the authors have contributed equally. 16. Syed R, Kantipudi R. New validated reverse-phase ultra-
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