• ATOLE D Department of Pharmaceutical Chemistry, Government College of Pharmacy, Aurangabad, Maharashtra, India.
  • DEOKATE UA Department of Pharmaceutical Chemistry, Government College of Pharmacy, Aurangabad, Maharashtra, India.


For pharmaceutical industry, HPLC is more than 40 years old. Even after instrumental advancements in HPLC problems still arise, troubleshooting still comes. HPLC is the main stay of analytical section nowadays in pharmaceutical industry whether it is API industry or formulation plant. Best approach for troubleshooting HPLC problems is going with systematic way. At the start of quantitative chromatographic analysis, the first parameter of paramount importance is baseline, especially for measuring the area of any given peak. Baseline usually suffers from these errors, namely, high baseline drift, periodic baseline fluctuation, and spikes. This article aims at suggesting some working methodologies which will resolve as well as avoid cases of such errors and failures.

Keywords: HPLC, Troubleshooting, Baseline, Pharmaceutical industry

Author Biography

DEOKATE UA, Department of Pharmaceutical Chemistry, Government College of Pharmacy, Aurangabad, Maharashtra, India.

Associate Professor at Government College Of Pharmacy


1. International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use. Validation of Analytical Procedures: Text and Methodology (Q2R1) Geneva, Switzerland: International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use; 2005.
2. Bridwell H, Dhingra V, Peckman D, Roark J, Lehman T. Perspectives on method validation: Importance of adequate method validation. Qual Assur J 2010;13:72-7.
3. Gorog S. The changing face of pharmaceutical analysis. Trends Anal Chem 2007;26:12-7.
4. Scott PW. Liquid Chromatography for the Analyst. New York: Marcel Dekker Inc.;1994. p. 1-10.
5. Stanley BJ, Foster CR, Guiochon G. On the reproducibility of column performance in liquid chromatography and the role of the packing density. J Chromatogr A 1997;761:41-51.
6. Christain GD. Analytical Chemistry. 6th ed. United States: John Wiley & Sons Inc.; 2001.
7. Nledner W, Karsten M, Stelner F, Swart R. Automating Method Development with an HPLC System Optimized for Scouting of Columns, Eluents and Other Method Parameters. United States; Pittcon Presentation; 2008.
8. Kar A. Pharmaceutical Drug Analysis. 1st ed. Tamil Nadu: New Age International Private Limited; 2001. p. 565-92.
9. Skoog DA, West DM, Holler FJ, Crouch SR. Fundamentals of Analytical Chemistry. 8th ed. Tamil Nadu: Brooks/Cole; 2004. p. 973.
10. Scott RP. Principles and Practice of Chromatography (Chrom-Ed Book Series). New York: Reese-Scott Partnership; 2003. p. 1-2.
11. Chatwal GR, Anand SK. Instrumental Methods of Chemical Analysis. 5th ed. Uttar Pradesh: Krishna Prakashan Media (P) Ltd.; 2004.
12. U.S. Food and Drug Administration. Title 21 Code of Federal Regulations (21 CFR Part 211.194(a)(2)). Laboratory Records. Available from: cfcfr/CFRSearch.cfm?fr=211.194. [Last accessed on 2009 Dec 11].
13. Chervet JP, Ursem M, Salzmann JP. Instrumental requirements for nano-scale liquid chromatography. Anal Chem 1996;68:1507-12.
14. Saito Y, Jinno K, Greibrokk T. Capillary columns in liquid chromatography: Between conventional columns and microchips. J Sep Sci 2004;27:1379-90.
15. Hernandez-Borges JH, Aturki Z, Rocco A, Fanali S. Recent applications in nano-liquid chromatography. J Sep Sci 2007;30:1589-610.
16. Qing H. Integrated Nano Liquid Chromatography System on a Chip, Ph.D. Thesis. California, USA: California Institute of Technology Pasadena; 2005.
17. Tzanavaras PD, Themelis DG. Review of recent applications of flow injection spectrophotometry to pharmaceutical analysis. Anal Chim Acta 2007;588:1-9.
18. Hassan BA. HPLC uses and importance in the pharmaceutical analysis and industrial field. Pharm Anal Acta 2012;3:9.
19. Gumustas M, Kurbanoglu S, Uslu B, Ozkan S. UPLC versus HPLC on drug analysis: Advantageous, applications and their validation parameters. Chromatographia 2013;76:21-2.
20. Glajch JL. Impact of new columns on drug development. Chromatogr Online 2014;32:36-41.
21. Thammana M. A review on high-performance liquid chromatography. Res Rev 2016;5:22.
22. Malviya R, Bansal V, Pal OP, Sharma P. High-performance liquid chromatography: A short review. J Glob Pharm Technol 2010;2:22-6.
23. Bokai M, Xin-Lei G, Xin-Ying Z. Application of high performance liquid chromatography in food and drug safety analysis. J Food Saf Qual 2016;7:4295-8.
24. Salgado P, Visnevschi-Necrasov T, Kiene RP, Azevedo I, Rocha AC, Almeida CM, et al. Determination of 3-mercaptopropionic acid by HPLC: A sensitive method for environmental applications. J Chromatogr B 2015;992:103-8.
25. Bird IM. High-performance liquid chromatography: Principles and clinical applications. Br Med J 1989;299:783-7.
26. Rogatsky E. Modern high performance liquid chromatography and HPLC 2016 international symposium. J Chromatogr Sep Tech 2016;7:135.
27. Wu N, Clausen AM. Fundamental and practical aspects of ultrahigh pressure liquid chromatography for fast separations. J Sep Sci 2007;30:1167.
28. Kirkland JJ. Modern Practice of Liquid Chromatography. New York: Wiley-Inter Science; 1971.
29. Crowther JB, Lauwers W, Adusumalli S, Shenbagamurthi P. The laboratory analyst’s role in the drug development process. In: Miller JM, Crowther JB, editors. Analytical Chemistry in a GMP Environment. A Practical Guide. New York: John Wiley; 2000. p. 448-9.
30. Dong MW. Modern HPLC for Practicing Scientists. Ch. 5. United States: John Wiley & Sons Publication; 2006. p. 111-6.
31. Prathap B, Deyb A, Raoa GH, Johnsona P, Arthanariswaranc P. A review-importance of RP-HPLC in analytical method development. Int J Novel Trend Pharm Sci 2013;3:15-23.
32. Available from: hplc-market-in-india-growing.html.
33. Available from: vaccine_quality/risk_july_2013.
34. Available from:
35. Academy Savant. Troubleshooting in HPLC, CLC-70, (Computer-based Instruction). Fullerton, CA: Academy Savant; 2020.
36. McNaught AD, Wilkinson A. Compendium of Chemical Terminology. 2nd ed. Oxford: Blackwell Scientific Publications, IUPAC; 1997.
37. Available from: HPLC-Gradient-Elution-Baseline-Drift.html.
38. Available from:
39. Available from: Troubleshooting.pdf.
40. Available from:
41. Available from:
42. Available from: Final%20TIPS%20and%20Tricks%20HPLC%20Troubleshooting%20 (2).pdf.
43. Available from: documents/a8cb7bc3-eb65-4244-8910-faeb1818df8c.pdf.
44. Available from:
45. Filgueiraa MR, Castells CB, Carra PW. A simple robust orthogonal background correction method for two-dimensional liquid chromatography. Anal Chem 2012:84;6747-52.
46. Snyder LR, Kirkland JJ, Dolan JW. Introduction to Modern Liquid Chromatography. United States: Wiley; 2009.
47. Neue UD, Corperation W. HPLC Troublr Shooting Guide. Available from: Waters [Last accessed 2020 Feb 20].
48. Available from: Detectors/det_nise.html.
49. Available from: EsHiemBpdmBlIEcChBgC&subNav=pmxgzFsHiemBpdmBlIEcCh BgCdD.
50. Mcnair H, Polite LN. Troubleshooting in high performance liquid chromatography. Sep Sci Technol 2007;8:459-77.
51. Nelson MD, Wolan JD. UV detector noise. LC GC 1999;17:12-5.
52. Available from: diagnosing-troubleshooting-hplc.html.
53. Available from: Detectors/det_nise.html.
54. Available from: diagnosing-troubleshooting-hplc.html.
55. Burghate AV. A review on HPLC-trouble shooting guide. Int J Pharm Sci Rev 2014;27:200-9.
56. Snyder LR, Kirkland JJ, Glajch JL. Practical HPLC Method Development. 2nd ed. New York: Wiley-Inter Science; 1997.
57. Wolan JD. LC user survey: Detector problem. LC GC1986;4:526-9.
58. Sadek PC. Troubleshooting HPLC Systems: A Bench Manual. New York: John Wiley & Sons; 2000.
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How to Cite
D, A., and D. UA. “INSIGHT INTO BASELINE TROUBLES AND PROBLEM RESOLUTIONS IN HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 13, no. 7, June 2020, pp. 35-38, doi:10.22159/ajpcr.2020.v13i7.37336.
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