I am working on the characterization of plant extracts using RP-HPLC. The extraction solvents used are methanol and ethanol. Kindly guide me about the standard sample preparation techniques commonly employed for RP-HPLC analysis.
Most of the instrument manufacturers will have a set of SOPs for extraction and preparation of biological materials and would be an easy way to start. Alternatively, one of the best ways to find a good method is to read some of the papers from the top people in the field, examples of which would be
Sonja Sturm's group from Vienna e.g.
1: Högner C, Sturm S, Seger C, Stuppner H. Development and validation of a rapid ultra-high performance liquid chromatography diode array detector method for Vitex agnus-castus. J Chromatogr B Analyt Technol Biomed Life Sci. 2013 May 15;927:181-90. doi: 10.1016/j.jchromb.2013.02.037.
2: Cheilari A, Sturm S, Intelmann D, Seger C, Stuppner H. Head-to-Head Comparison of Ultra-High-Performance Liquid Chromatography with Diode Array Detection versus Quantitative Nuclear Magnetic Resonance for the Quantitative Analysis of the Silymarin Complex in Silybum marianum Fruit Extracts. J Agric Food Chem. 2016 Feb 24;64(7):1618-26. doi: 10.1021/acs.jafc.5b05494.
3. Assimopoulou AN, Sturm S, Stuppner H, Papageorgiou VP. Preparative isolation and purification of alkannin/shikonin derivatives from natural products by high-speed counter-current chromatography. Biomed Chromatogr. 2009 Feb;23(2):182-98. doi: 10.1002/bmc.1101.
Rob Verpoorte e.g.
1: Pan Q, Saiman MZ, Mustafa NR, Verpoorte R, Tang K. A simple and rapid HPLC-DAD method for simultaneously monitoring the accumulation of alkaloids and precursors in different parts and different developmental stages of Catharanthus roses plants. J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Mar 1;1014:10-6. doi:
10.1016/j.jchromb.2016.01.034.
2: El-Seedi HR, El-Ghorab DM, El-Barbary MA, Zayed MF, Göransson U, Larsson S, Verpoorte R. Naturally occurring xanthones; latest investigations: isolation, structure elucidation and chemosystematic significance. Curr Med Chem. 2009;16(20):2581-626. Review. PubMed PMID: 19601799.
Oliver Fiehn e.g.
Ikegami T, Horie K, Saad N, Hosoya K, Fiehn O, Tanaka N. Highly efficient
analysis of underivatized carbohydrates using monolithic-silica-based capillary hydrophilic interaction (HILIC) HPLC. Anal Bioanal Chem. 2008 Aug;391(7):2533-42.
doi: 10.1007/s00216-008-2060-6.
Also manu Chinese and Brazillian groups have long histories in separation of natural products e.g.
Lv H, ut al J Chromatogr Sci. 2016 May-Jun;54(5):744-51. doi:
10.1093/chromsci/bmv249. Epub 2016 Feb 3.
Chen X, Wang J, Hu S, Bai X. J Sep Sci. 2016 Mar;39(5):827-34. doi: 10.1002/jssc.201501160.
Rozhon W, et al . Phytochem Anal. 2018 Jan;29(1):48-58. doi: 10.1002/pca.2713.
de Mesquita ML, et al Pharmacogn Mag. 2015 Oct;11(Suppl 2):S316-21. doi: 10.4103/0973-1296.166062.
Check this review:
https://www.researchgate.net/publication/6672624_Techniques_of_Preparing_Plant_Material_for_Chromatographic_Separation_and_Analysis
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