UHPLC Technology: Recent Developments and Emerging Applications in Analytical Chemistry

Abhishek Chauhan; Ankit Sharma; Amar Deep Ankalgi; Nikhil Sharma; Mahendra Singh Ashawat

doi:10.52711/2231-5675.2025.00050

Asian Journal of Pharmaceutical Analysis

ISSN

2231-5675 (Online)
2231-5667 (Print)

Submit Article

UHPLC Technology: Recent Developments and Emerging Applications in Analytical Chemistry

Author(s): Abhishek Chauhan, Ankit Sharma, Amar Deep Ankalgi, Nikhil Sharma, Mahendra Singh Ashawat

Email(s): ankitsharma.0506@gmail.com

DOI: 10.52711/2231-5675.2025.00050

Address: Abhishek Chauhan, Ankit Sharma*, Amar Deep Ankalgi, Nikhil Sharma, Mahendra Singh Ashawat
Department of Pharmaceutical Analysis, Laureate Institute of Pharmacy, Kathog, Jawalamukhi 176031, Himachal Pradesh, India.
*Corresponding Author

Published In: Volume - 15, Issue - 4, Year - 2025

View HTML

Purchase PDF

ABSTRACT:
UPLC is a contemporary method that offers liquid chromatography a new path. UPLC, or ultra-performance liquid chromatography, improves primarily in three areas: sensitivity, speed, and resolution. High-performance liquid chromatography (HPLC) lacks the resolution, speed, and sensitivity that ultra-performance liquid chromatography (UPLC) provides for particles smaller than 2µm in diameter. In this work, the current status, benefits, and drawbacks of UHPLC in technique development are critically examined. In UPLC, separation and quantification are carried out at extremely high pressures (up to 100M Pa). When compared to HPLC, it is found that high pressure has no detrimental effects on the analytical column. Additionally, UPLC uses less time and solvent. Application of UHPLC in academic institutions and the pharmaceutical industry has been successfully used for compound identification and determination. Using examples of some of the most cutting-edge work in pharmaceutical analysis, the author of this review paper gives information about UHPLC and its application.

Keywords:

Cite this article:
Abhishek Chauhan, Ankit Sharma, Amar Deep Ankalgi, Nikhil Sharma, Mahendra Singh Ashawat. UHPLC Technology: Recent Developments and Emerging Applications in Analytical Chemistry. Asian Journal of Pharmaceutical Analysis. 2025; 15(4):319-7. doi: 10.52711/2231-5675.2025.00050

Cite(Electronic):
Abhishek Chauhan, Ankit Sharma, Amar Deep Ankalgi, Nikhil Sharma, Mahendra Singh Ashawat. UHPLC Technology: Recent Developments and Emerging Applications in Analytical Chemistry. Asian Journal of Pharmaceutical Analysis. 2025; 15(4):319-7. doi: 10.52711/2231-5675.2025.00050 Available on: https://ajpaonline.com/AbstractView.aspx?PID=2025-15-4-12

REFERNCES:
1. Karayannis MI, Efstathiou CE. Significant steps in the evolution of analytical chemistry—Is the today’s analytical chemistry only chemistry. Talanta. 2012 Dec 15; 102:7-15.
2. Perkel JM. Advances in Analytical Chemistry: Processes, Techniques, and Instrumentation.
3. Van Deemter JJ, Zuiderweg FJ, Klinkenberg AV. Longitudinal diffusion and resistance to mass transfer as causes of nonideality in chromatography. Chemical Engineering Science. 1956 Sep 1;5(6):271-89.
4. Gupta MK, Ghuge A, Parab M, Al-Refaei Y, Khandare A, Dand N, Waghmare N. A comparative review on high-performance liquid chromatography (HPLC), ultra-performance liquid chromatography (UPLC) & high-performance thin layer chromatography (HPTLC) with current updates. Current Issues in Pharmacy and Medical Sciences. 2022; 35(4): 224-8.
5. Wu N, Lippert JA, Lee ML. Practical aspects of ultrahigh pressure capillary liquid chromatography. Journal of Chromatography A. 2001 Mar 9;911(1):1-2.
6. Jebaliya H, Patel M, Jadeja Y, Dabhi B, Shah A. A comparative validation study of fluconazole by HPLC and UPLC with forced degradation study. Chromatography Research International. 2013; 2013(1): 673150.
7. Walter TH, Andrews RW. Recent innovations in UHPLC columns and instrumentation. TrAC Trends in Analytical Chemistry. 2014 Dec 1; 63: 14-20.
8. Patil A. A Review on Ultra Performance Liquid Chromatography (UPLC). Asian Journal of Pharmaceutical Technology & Innovation. 2015; 10: 86-96.
9. Basharat R, Kotra V, Loong LY, Mathews A, Kanakal MM, Devi CB, Nyamathulla S, Varala R, Ming LC, Rao KS, Babu BH. A mini-review on ultra-performance liquid chromatography. Oriental Journal of Chemistry. 2021; 37(4): 847.
10. Khan H, Ali J. UHPLC: Applications in pharmaceutical analysis. Asian J. Pharm. Ana. 2017 May; 7(2): 124-31.
11. Patil VP, Tathe RD, Devdhe SJ, Angadi SS, Kale SH. Ultra-performance liquid chromatography: A review. International Research Journal of Pharmacy. 2011; 2(6): 39-44.
12. Patil DR, Patil KP, Patil SA. A Review on UHPLC Method. Research Journal of Pharmacology and Pharmacodynamics. 2024; 16(2): 123-6.
13. Sheliya KG, Shah KV. Ultra-Performance Liquid Chromatography (Uplc): A Modern Chromatography Technique. Pharma Science Monitor. 2013 Apr 15;4(3)..
14. Bharatbhai PN, Surati J, Akbari A, Patel S, Shah K, Solanki D. A Review on UHPLC Instrumentation with Advancements. J. Adv. Pharm. Sci. 2023; 1:33-44.
15. Rathod RH, Chaudhari SR, Patil AS, Shirkhedkar AA. Ultra-high performance liquid chromatography-MS/MS (UHPLC-MS/MS) in practice: analysis of drugs and pharmaceutical formulations. Future Journal of Pharmaceutical Sciences. 2019 Dec; 5:1-26.
16. Taleuzzaman M, Ali S, Gilani SJ, Imam SS, Hafeez A. Ultra-performance liquid chromatography (UPLC)-a review. Austin J Anal Pharm Chem. 2015;2(6):1056.
17. Chen S, Kord A. Theoretical and experimental comparison of mobile phase consumption between ultra-high-performance liquid chromatography and high-performance liquid chromatography. Journal of Chromatography A. 2009 Aug 21; 1216(34):6204-9.
18. Behnoush B, Sheikhazadi A, Bazmi E, Fattahi A, Sheikhazadi E, Anary SH. Comparison of UHPLC and HPLC in Benzodiazepines Analysis of Postmortem Samples: A Case–Control Study. Medicine. 2015 Apr 1; 94(14): e640.
19. Narwate NB, PJ GP, AV GA, AS DA, JG WJ. Ultra-performance liquid chromatography: A new revolution in liquid chromatography. International Journal of Pharmaceutics and Drug Analysis. 2014; 2(1): 25-34.
20. Samatha Y, Srividya A, Ajitha A, Rao VU. Ultra-performance liquid chromatography (UPLC). World J Pharm Pharm Sci. 2015;4(6):356-67.
21. Chawla G, Ranjan C. Principle, instrumentation, and applications of UPLC: A novel technique of liquid chromatography. Open Chemistry Journal. 2016 May 6; 3(1): 1-6.
22. Sharma DK, Kim SG, Lamichhane R, Lee KH, Poudel A, Jung HJ. Development of UPLC fingerprint with multi-component quantitative analysis for quality consistency evaluation of herbal medicine “Hyangsapyeongwisan”. Journal of Chromatographic Science. 2016 Apr 1;54(4):536-46.
23. Kumar A, Saini G, Nair A, Sharma R. UPLC: a preeminent technique in pharmaceutical analysis. Acta Pol Pharm. 2012 Jun; 69(3): 371-80.
24. Dong MW, Zhang K. Ultra-high-pressure liquid chromatography (UHPLC) in method development. TrAC Trends in Analytical Chemistry. 2014 Dec 1; 63: 21-30.
25. Kashid AM, Polshettiwar PP, Bhosale KM. Analytical method development and validation of teriflunomide active pharmaceutical ingredient by RP-UHPLC. Asian Journal of Pharmaceutical Analysis. 2022; 12(3):166-72.
26. Yu HC, Pan CW, Xie QP, Zheng Y, Hu YZ, Lin YM. Simultaneous determination of tedizolid and linezolid in rat plasma by ultra-performance liquid chromatography tandem mass spectrometry and its application to a pharmacokinetic study. Journal of Chromatography B. 2016 Feb 1; 1011: 94-8.
27. Wang XS, Sun JC, Yang RN, Ren J, Liang S, Sun M, Wang YF, Gao SG. Determination of cephalomannine in rat plasma by gradient elution UPLC–MS/MS method. Journal of Chromatography B. 2014 Jul 15; 963: 70-4.
28. Rezk MR, Bendas ER, Basalious EB, Karim IA. Development and validation of sensitive and rapid UPLC–MS/MS method for quantitative determination of daclatasvir in human plasma: Application to a bioequivalence study. Journal of Pharmaceutical and Biomedical Analysis. 2016 Sep 5; 128: 61-6.
29. Li TX, Chu CS, Zhu JY, Yang TY, Zhang J, Hu YT, Yang XH. A rapid UPLC‐MS/MS method for the determination of oleanolic acid in rat plasma and liver tissue: application to plasma and liver pharmacokinetics. Biomedical Chromatography. 2016 Apr; 30(4): 520-7.
30. Kormány R, Rácz N, Fekete S, Horváth K. Development of a fast and robust UHPLC method for apixaban in-process control analysis. Molecules. 2021 Jun 8; 26(12):3505.
31. Al‐Quadeib BT, Radwan MA, Siller L, Mutch E, Horrocks B, Wright M, Alshaer A. Therapeutic monitoring of amphotericin B in Saudi ICU patients using UPLC MS/MS assay. Biomedical Chromatography. 2014 Dec; 28(12):1652-9.
32. Szeitz A, Edginton AN, Peng HT, Cheung B, Riggs KW. A validated enantioselective assay for the determination of ibuprofen in human plasma using ultra performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). American Journal of Analytical Chemistry. 2010 Sep 14; 1(2):47-58.
33. Bhatt M, Shah S, Shivprakash. Rapid ultraperformance liquid chromatography–tandem mass spectrometry method for quantification of oxcarbazepine and its metabolite in human plasma. Biomedical Chromatography. 2011 Jul; 25(7):751-9.
34. Cao G, Zhang Q, Yang X. Determination of letrozole in rat plasma by UPLC-MS/MS and its application to a pharmacokinetic study. Latin American Journal of Pharmacy. 2015; 34(1):45-50.
35. Wang S, Wu H, Geng P, Lin Y, Liu Z, Zhang L, Ma J, Zhou Y, Wang X, Wen C. Pharmacokinetic study of dendrobine in rat plasma by ultra‐performance liquid chromatography tandem mass spectrometry. Biomedical Chromatography. 2016 Jul; 30(7):1145-9.
36. Chen Y, Zhang S, Chen D, Zhou M, Zheng J, Xiang Z. An UPLC-MS/MS method for determination of solasonine in rat plasma and its application of a pharmacokinetic and bioavailability study. Journal of Chromatography B. 2015 Mar 15; 985:1-5.
37. Patil Vedashri. G, Patil Tejaswini. R, Dhankani Mansi. A, Amitkumar R Dhankani, Sunil P Pawar. A Review on Ultra High Performance liquid Chromatography Method. International Journal for Research in Applied Science and Engineering Technology (IJRASET). 11(6): 311-319.
38. Khan H, Ali M, Ahmad S, Ahmad N, Ahuja A, Baboota S, Ali J. Validated UPLC/Q-TOF-MS method for simultaneous determination of aceclofenac, paracetamol, and their degradation products in tablets. Journal of Liquid Chromatography & Related Technologies. 2012 Jan 1;35(1):109-28.
39. Rathod RH, Chaudhari SR, Patil AS, Shirkhedkar AA. Ultra-high performance liquid chromatography-MS/MS (UHPLC-MS/MS) in practice: analysis of drugs and pharmaceutical formulations. Future Journal of Pharmaceutical Sciences. 2019 Dec; 5:1-26.
40. Khan H, Ali M, Ahuja A, Ali J. Validated HPLC-UV method for simultaneous determination of some anti-inflammatory and analgesic drugs. Asian Journal of Pharmaceutical Analysis. 2016; 6(3): 183-7.
41. Khan H, Ali M, Ahuja A, Ali J. Application of Validated UPLC/Q-TOF-MS Method for Simultaneous Determination of Telmisartan, Hydrochlorothiazide and their Degradation Products in Tablets. Asian Journal of Research in Pharmaceutical Science. 2017; 7(2): 105-11.
42. Khan H. Identification and characterization of degradation products of valsartan by UPLC/q-tof-MS technique. Asian Journal of Pharmaceutical Research. 2021; 11(1): 1-5.
43. Karunakaran K, Navaneethan G, Elango KP. Development of a new RP-UPLC method for the determination of rabeprazole sodium in pharmaceutical formulation and application in dissolution studies. Tropical Journal of Pharmaceutical Research. 2011; 10(5): 655-61.
44. Khan H, Ali J. Formulation and In-Vitro Evaluation of a Bilayer Matrix Tablet Containing Aceclofenac as Sustained Release and Paracetamol as Immediate Release. Drug Delivery Letters. 2014 Dec 1; 4(3): 221-6.
45. Khan H, Ali M, Ahuja A, Ali J. Validated UPLC/QTOF-MS Method for Simultaneous Determination of Aceclofenac, Paracetamol and Chlorzoxazone in Human Plasma and its Application to Pharmacokinetic Study. Asian J. Pharm. Ana. 2017 Jun 19; 7(2): 93-9.
46. Johnson KA, Plumb R. Investigating the human metabolism of acetaminophen using UPLC and exact mass oa-TOF MS. Journal of Pharmaceutical and Biomedical Analysis. 2005 Sep 15; 39(3-4): 805-10.
47. Xie G, Plumb R, Su M, Xu Z, Zhao A, Qiu M, Long X, Liu Z, Jia W. Ultra‐performance LC/TOF MS analysis of medicinal Panax herbs for metabolomic research. Journal of Separation Science. 2008 Apr; 31(6‐7): 1015-26.
48. Li L, Luo GA, Liang QL, Hu P, Wang YM. Rapid qualitative and quantitative analyses of Asian ginseng in adulterated American ginseng preparations by UPLC/Q-TOF-MS. Journal of Pharmaceutical and Biomedical Analysis. 2010 May 1; 52(1): 66-72.
49. Zhang HM, Li SL, Zhang H, Wang Y, Zhao ZL, Chen SL, Xu HX. Holistic quality evaluation of commercial white and red ginseng using a UPLC-QTOF-MS/MS-based metabolomics approach. Journal of Pharmaceutical and Biomedical Analysis. 2012 Mar 25; 62: 258-73.
50. Mazzeo J, Wheat T, Gillece-Castro B, ZILING L. Next generation peptide mapping with ultra-performance liquid chromatography. Biopharm International. 2006; 19(1): 22-30.
51. IS K, Wheat T. Current Applications of UHPLC in Biotechnology Today, Part I (Peptide Mapping and Amino Acid Analysis).
52. Wheat TE, Krull IS, Rathore A. Current applications of UHPLC in biotechnology, Part II: Proteins and Glycans. LC-GC North America. 2011 Dec 1; 29(12): 1052-61.
53. Ren XN, Yin SA, Yang ZY, Yang XG, Shao B, Ren YP, Zhang J. Application of UPLC-MS/MS method for analyzing B-vitamins in human milk. Biomedical and Environmental Sciences. 2015 Oct 1; 28(10): 738-50.
54. Ortega N, Romero MP, Macià A, Reguant J, Anglès N, Morelló JR, Motilva MJ. Comparative study of UPLC–MS/MS and HPLC–MS/MS to determine procyanidins and alkaloids in cocoa samples. Journal of Food Composition and Analysis. 2010 May 1; 23(3): 298-305.
55. Ibáñez M, Guerrero C, Sancho JV, Hernández F. Screening of antibiotics in surface and wastewater samples by ultra-high-pressure liquid chromatography coupled to hybrid quadrupole time-of-flight mass spectrometry. Journal of Chromatography A. 2009 Mar 20; 1216(12): 2529-39.
56. Thomas T, Petrie K, Shim J, Abildskov KM, Westhoff CL, Cremers S. A UPLC-MS/MS method for therapeutic drug monitoring of etonogestrel. Therapeutic Drug Monitoring. 2013 Dec 1; 35(6): 844-8.
57. Khan H. Uhplc/q-tof-ms method for determination of antihypertensive drugs and its application to pharmacokinetic study. Asian Journal of Pharmaceutical Analysis. 2021; 11(1): 22-6.
58. Kivrak Ş, Göktürk T, Kivrak I, Kaya E, Karababa E. Investigation of phenolic profiles and antioxidant activities of some Salvia species commonly grown in Southwest Anatolia using UPLC-ESI-MS/MS. Food Science and Technology. 2018 Apr 19; 39(2): 423-31.
59. Picó Y, la Farré M, Soler C, Barceló D. Identification of unknown pesticides in fruits using ultra-performance liquid chromatography–quadrupole time-of-flight mass spectrometry: Imazalil as a case study of quantification. Journal of Chromatography A. 2007 Dec 28; 1176(1-2): 123-34.
60. Badoud F, Grata E, Perrenoud L, Avois L, Saugy M, Rudaz S, Veuthey JL. Fast analysis of doping agents in urine by ultra-high-pressure liquid chromatography–quadrupole time-of-flight mass spectrometry: I. Screening analysis. Journal of Chromatography A. 2009 May 15; 1216(20): 4423-33.
61. Díaz R, Ibáñez M, Sancho JV, Hernández F. Qualitative validation of a liquid chromatography–quadrupole-time of flight mass spectrometry screening method for organic pollutants in waters. Journal of Chromatography A. 2013 Feb 8; 1276: 47-57.
62. Khan H. UHPLC: Development and Recent Applications. Asian Journal of Pharmaceutical Analysis. 2024 Mar 8; 14(1): 39-46