Abstract
Preparative HPLC (prep-HPLC) has emerged as a mainstay of natural products isolation and purification. The various modes available to date, e.g. normal phase, reversed-phase, size exclusion, and ion exchange, can be used to purify most classes of natural products. This chapter presents an overview of the different modes along with a practical guide as to how to purify a natural product using the most robust and widely used of the modes, namely reversed-phase prep-HPLC. Instrumentation set-up and detection methods, sample preparation, method development, and sample work up are also discussed.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Eurby MR, Petersson P (2003) Chromatographic classification and comparison of commercially available reversed-phase liquid chromatographic columns using principle component analysis. J Chromatog A 994:13–26
Hostettman K, Hostettman M, Marston A (1986) Preparative chromatography techniques: applications in natural product isolation. Springer, Berlin, Germany, pp 37–39
Young CS, Dolan JW (2003) Success with evaporative light-scattering detection. LC-GC Europe 13:132–137
Snyder LR, Kirkland JJ, Glajch JL (1997) Practical HPLC method development, 2nd edn. Wiley, New York, USA
Dolan JW (2000) Starting out right, Part 3 – The role of the solvent in controlling selectivity. LC-GC Europe 13:148–156
Dolan JW (2000) Starting out right, Part 6 – The scouting gradient alternative. LC-GC Europe 13:388–394
Dolan JW (2003) How much is enough? LC-GC Europe 16:740–745
Neue UD (1997) HPLC Columns. Wiley VCH, NY, USA
Mazzei JL, d’Avila LA (2003) Chromatographic models as tools for scale-up of isolation of natural products by semi-preparative HPLC. J Liq Chromatog and Related Technol 26:177–193
Zhang M, Stout MJ, Kubo I (1992) Isolation of ecdysteroids from Vitex strickeri using RLCC and recycling HPLC. Phytochemistry 31:247–250
Delazar A, Khodaie L, Afshar J, Nahar L, Sarker SD (2010) Isolation and free-radical-scavenging properties of cyaniding 3-O-glycosides from the fruits of Ribes bibersteinii Berl. Acta Pharmaceutica 60:1–11
Nazemiyeh H, Kazemi EM, Zare K, Jodari M, Nahar L, Sarker SD (2010) Free radical scavengers from the aerial parts of Euphorbia petiolata. J Nat Med 64:187–190
Suggested Readings
Basic liquid chromatography (1996–2000). Yuri Kazakevich, Harold McNair on http://hplc.chem.shu.edu/NEW/HPLC_Book/index.html
Hostettman K, Hostettman M, Marston A (1986) Preparative chromatography techniques: applications in natural product isolation. Springer, Berlin, Germany
Shelley PR (1996) High performance liquid chromatography. In: Verall MS (ed) Downstream processing of natural products. Wiley, Chichester, UK
Katz ED (ed) (1996) High pressure liquid chromatography: principles and methods and biotechnology. Wiley, Chichester, UK
Dolan JW (ed) (2004) LC troubleshooting. In: LC-GC Europe. Advanstar, US. Also found at http://www.lcgceurope.com
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Latif, Z., Sarker, S.D. (2012). Isolation of Natural Products by Preparative High Performance Liquid Chromatography (Prep-HPLC). In: Sarker, S., Nahar, L. (eds) Natural Products Isolation. Methods in Molecular Biology, vol 864. Humana Press. https://doi.org/10.1007/978-1-61779-624-1_10
Download citation
DOI: https://doi.org/10.1007/978-1-61779-624-1_10
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-623-4
Online ISBN: 978-1-61779-624-1
eBook Packages: Springer Protocols