Chromatographia

, Volume 80, Issue 8, pp 1143–1159 | Cite as

A 14 Parameter Study of UHPLC’s for Method Development Transfer and Troubleshooting

  • Imad A. Haidar Ahmad
  • Frank Hrovat
  • Arianne Soliven
  • Adrian Clarke
  • Paul Boswell
  • Thomas Tarara
  • Andrei Blasko
Original
First Online:
Received:
Revised:
Accepted:

Abstract

Five ultra-high pressure liquid chromatography (UHPLC) instruments were compared to one another by examining the overall system performance and key functions of the system parts including: pump, auto-sampler, thermo-stated column compartment, and the detector. The five UHPLC systems used in this study were: ThermoFisher Vanquish, Agilent 1290 Infinity I, Agilent 1290 Infinity II, Waters Acquity I-Class, and Shimadzu Nexera X2. The identities of the systems were blinded in the results and discussion section to use this study for scientific purposes only rather than for competition and marketing. The following tests were performed to evaluate and compare the five UHPLC systems: injector linearity and precision, sample carryover, sample (autosampler) temperature accuracy, column temperature accuracy and precision, pressure ripple, pump mixing accuracy, flow rate accuracy, detector drift and noise, detector linearity, wavelength accuracy, extra-column volume, and dwell volume determination. This study presents an approach on how to test the performance of UHPLC systems along with potential problems that analysts may face when using the UHPLC systems, examples of such issues are: retention time irreproducibility, low sensitivity, method transfer failure, etc.

Keywords

UHPLC UPLC Ultra-high pressure liquid chromatography Mixing accuracy Mixing precision Temperature accuracy and precision Baseline noise Baseline drift Detector linearity Injector linearity Wavelength accuracy Dwell volume, and extracolumn volume Sample temperature Pressure ripple Flow rate accuracy Flow rate precision 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgements

The authors would like to thank the Vendors (Agilent, Shimadzu, ThermoFisher, and Waters) for the provision of loan instruments and scientific advice.

Compliance with Ethical Standards

Conflict of interest

The author declares that he has no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by the authors.

Funding

No Funding for this work was provided.

References

  1. 1.
    De VJ, Broeckhoven K, Eeltink S (2016) Advances in ultrahigh-pressure liquid chromatography technology and system design. Anal Chem 88:262–278CrossRefGoogle Scholar
  2. 2.
    Aasberg D, Lesko M, Samuelsson J, Kaczmarski K, Fornstedt T (2014) Method transfer from high-pressure liquid chromatography to ultra-high-pressure liquid chromatography. I. A thermodynamic perspective. J Chromatogr A 1362:206–217CrossRefGoogle Scholar
  3. 3.
    McCalley DV (2014) The impact of pressure and frictional heating on retention, selectivity and efficiency in ultra-high-pressure liquid chromatography. TrAC. Trends Anal Chem 63:31–43CrossRefGoogle Scholar
  4. 4.
    Novakova L, Veuthey JL, Guillarme D (2011) Practical method transfer from high performance liquid chromatography to ultra-high performance liquid chromatography: the importance of frictional heating. J Chromatogr A 1218:7971–7981CrossRefGoogle Scholar
  5. 5.
    Boswell PG, Schellenberg JR, Carr PW, Cohen JD, Hegeman AD (2011) Easy and accurate high-performance liquid chromatography retention prediction with different gradients, flow rates, and instruments by back-calculation of gradient and flow rate profiles. J Chromatogr A 1218:6742–6749CrossRefGoogle Scholar
  6. 6.
    Dolan JW (2006) Dwell volume revisited. LCGC North Am 24:458–466Google Scholar
  7. 7.
    Schellinger AP, Carr PW (2005) A practical approach to transferring linear gradient elution methods. J Chromatogr A 1077:110–119CrossRefGoogle Scholar
  8. 8.
    Snyder LR, Dolan JW (2006) Method development. High-performance gradient elution. Wiley, New York, pp 74–132CrossRefGoogle Scholar
  9. 9.
    Gritti F, Felinger A, Guiochon G (2006) Influence of the errors made in the measurement of the extra-column volume on the accuracies of estimates of the column efficiency and the mass transfer kinetics parameters. J Chromatogr A 1136:57–72CrossRefGoogle Scholar
  10. 10.
    Gritti F, Guiochon G (2010) On the extra-column band-broadening contributions of modern, very high pressure liquid chromatographs using 2.1 mm I.D. columns packed with sub-2 μm particles. J Chromatogr A 1217:7677–7689CrossRefGoogle Scholar
  11. 11.
    Gritti F, Guiochon G (2014) Accurate measurements of the true column efficiency and of the instrument band broadening contributions in the presence of a chromatographic column. J Chromatogr A 1327:49–56CrossRefGoogle Scholar
  12. 12.
    Gritti F, McDonald T, Gilar M (2015) Impact of the column hardware volume on resolution in very high pressure liquid chromatography non-invasive investigations. J Chromatogr A 1420:54–65CrossRefGoogle Scholar
  13. 13.
    Walter TH, Andrews RW (2014) Recent innovations in UHPLC columns and instrumentation. TrAC Trends Anal Chem 63:14–20CrossRefGoogle Scholar
  14. 14.
    Gritti F, Guiochon G (2011) On the minimization of the band-broadening contributions of a modern, very high pressure liquid chromatograph. J Chromatogr A 1218:4632–4648CrossRefGoogle Scholar
  15. 15.
    Buckenmaier S, Miller CA, van de Goor T, Dittmann MM (2015) Instrument contributions to resolution and sensitivity in ultra high performance liquid chromatography using small bore columns: comparison of diode array and triple quadrupole mass spectrometry detection. J Chromatogr A 1377:64–74CrossRefGoogle Scholar
  16. 16.
    Fekete S, Kohler I, Rudaz S, Guillarme D (2014) Importance of instrumentation for fast liquid chromatography in pharmaceutical analysis. J Pharm Biomed Anal 87:105–119CrossRefGoogle Scholar
  17. 17.
    Dong MW, Zhang K (2014) Ultra-high-pressure liquid chromatography (UHPLC) in method development. TrAC. Trends Anal Chem 63:21–30CrossRefGoogle Scholar
  18. 18.
    Wahab MF, Dasgupta PK, Kadjo AF, Armstrong DW (2016) Sampling frequency, response times and embedded signal filtration in fast, high efficiency liquid chromatography: a tutorial. Anal Chim Acta 907:31–44CrossRefGoogle Scholar
  19. 19.
    Colgate SO, Berger TA (2015) On axial temperature gradients due to large pressure drops in dense fluid chromatography. J Chromatogr A 1385:94–102CrossRefGoogle Scholar
  20. 20.
    de Villiers A, Lauer H, Szucs R, Goodall S, Sandra P (2006) Influence of frictional heating on temperature gradients in ultra-high-pressure liquid chromatography on 2.1 mm I.D. columns. J Chromatogr A 1113:84–91CrossRefGoogle Scholar
  21. 21.
    Fallas MM, Hadley MR, McCalley DV (2009) Practical assessment of frictional heating effects and thermostat design on the performance of conventional (3 μm and 5 μm) columns in reversed-phase high-performance liquid chromatography. J Chromatogr A 1216:3961–3969CrossRefGoogle Scholar
  22. 22.
    Fekete S, Guillarme D (2015) Estimation of pressure-, temperature- and frictional heating-related effects on proteins’ retention under ultra-high-pressure liquid chromatographic conditions. J Chromatogr A 1393:73–80CrossRefGoogle Scholar
  23. 23.
    Fekete S, Guillarme D, Fekete J (2014) Estimation of the effects of longitudinal temperature gradients caused by frictional heating on the solute retention using fully porous and superficially porous sub-2 μm materials. J Chromatogr A 1359:124–130CrossRefGoogle Scholar
  24. 24.
    Fekete S, Veuthey J-L, Guillarme D (2015) Comparison of the most recent chromatographic approaches applied for fast and high resolution separations: theory and practice. J Chromatogr A 1408:1–14CrossRefGoogle Scholar
  25. 25.
    Fekete S, Veuthey J-L, McCalley DV, Guillarme D (2012) The effect of pressure and mobile phase velocity on the retention properties of small analytes and large biomolecules in ultra-high pressure liquid chromatography. J Chromatogr A 1270:127–138CrossRefGoogle Scholar
  26. 26.
    Grinias JP, Keil DS, Jorgenson JW (2014) Observation of enhanced heat dissipation in columns packed with superficially porous particles. J Chromatogr A 1371:261–264CrossRefGoogle Scholar
  27. 27.
    Nguyen DTT, Guillarme D, Rudaz S, Veuthey J-L (2006) Fast analysis in liquid chromatography using small particle size and high pressure. J Sep Sci 29:1836–1848CrossRefGoogle Scholar
  28. 28.
    Camenzuli M, Ritchie HJ, Shalliker RA (2013) Improving HPLC separation performance using parallel segmented flow chromatography. Microchem J 111:3–7CrossRefGoogle Scholar
  29. 29.
    Shalliker RA, Ritchie H (2014) Segmented flow and curtain flow chromatography: overcoming the wall effect and heterogeneous bed structures. J Chromatogr A 1335:122–135CrossRefGoogle Scholar
  30. 30.
    Soliven A, Foley D, Pereira L, Hua S, Edge T, Ritchie H, Dennis GR, Andrew Shalliker R (2014) Improving the performance of narrow-bore HPLC columns using active flow technology. Microchem J 116:230–234CrossRefGoogle Scholar
  31. 31.
    Karger BL, Martin M, Guiochon G (1974) Role of column parameters and injection volume on detection limits in liquid chromatography. Anal Chem 46:1640–1647CrossRefGoogle Scholar
  32. 32.
    Neue UD, Kele M (2007) Performance of idealized column structures under high pressure. J Chromatogr A 1149:236–244CrossRefGoogle Scholar
  33. 33.
    Haidar Ahmad IA (2017) Necessary analytical skills and knowledge for identifying, understanding, and performing HPLC troubleshooting. Chromatographia 80:705–730CrossRefGoogle Scholar
  34. 34.
    Abate-Pella D, Freund DM, Ma Y, Simón-Manso Y, Hollender J, Broeckling CD, Huhman DV, Krokhin OV, Stoll DR, Hegeman AD, Kind T, Fiehn O, Schymanski EL, Prenni JE, Sumner LW, Boswell PG (2015) Retention projection enables accurate calculation of liquid chromatographic retention times across labs and methods. J Chromatogr A 1412:43–51CrossRefGoogle Scholar
  35. 35.
    Thompson JW, Kaiser TJ, Jorgenson JW (2006) Viscosity measurements of methanol–water and acetonitrile–water mixtures at pressures up to 3500 bar using a novel capillary time-of-flight viscometer. J Chromatogr A 1134:201–209CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Imad A. Haidar Ahmad
    • 1
  • Frank Hrovat
    • 1
  • Arianne Soliven
    • 1
  • Adrian Clarke
    • 2
  • Paul Boswell
    • 3
  • Thomas Tarara
    • 1
  • Andrei Blasko
    • 1
  1. 1.Novartis Pharmaceuticals Corporation, Technical Research and Development, PHADSan CarlosUSA
  2. 2.Novartis Pharma AG, Chemical and Analytical Development, CHADBaselSwitzerland
  3. 3.Department of Horticultural ScienceUniversity of MinnesotaSt. PaulUSA

Personalised recommendations