Bioassay-coupled Chromatographic Analysis of Medicinal Natural Products: A  Review

Taiwo  O. Elufioye

Authors

Taiwo O. Elufioye Department of Pharmacognosy, Faculty of Pharmacy, University of Ibadan, Nigeria

Keywords:

Medicinal plants,, Bioassay,, Chromatographic analysis,, Hyphenated techniques,, Secondary metabolites

Abstract

Medicinal plants have found immense use in drug discovery over the years as several chemically available drugs today are either directly from plant secondary metabolites or have their templates from such. Recent advances in the application of bioassay-coupled chromatographic analysis of plant extracts have led to reliable characterization of unknown samples with regards to their activity profile as well as the compounds responsible for observed activity. This paper reviews the progress made in the field of bioassay-chromatography hyphenated techniques for natural products analysis, notably medicinal plants. Different bioassays including microbial, biochemical, micro-chemical and cell based assays as well as several chromatographic and spectroscopic techniques such as Thin Layer Chromatography (TLC), High Performance Thin Layer Chromatography (HPTLC), Liquid Chromatography (LC), Over-Pressured Layer Chromatography (OPLC), Gas Chromatography (GC), Nuclear Magnetic Resonance (NMR), Fourier Transform Infra-red (FTIR) spectroscopy, Mass Spectrometry (MS), Surface Enhanced Raman Spectroscopy (SERS), Diffuse Reflectance Infrared Fourier Transform (DRIFT), etc. are discussed in this paper.

References

Agatonovic-Kustrin S, Morton DW, Yusof AP. Thin layer Chromatography – Bioassay us powerful tool for rapid identification of bioactive components in botanical extracts. Mod. Chem. Appl. 2015; 3(1):1-2.

Sherma J. Bioluminescence detection in thin layer chromatography. Studia universitatis babes-bolyai, Chemia. 2009; 1(2).

Morlock GE. Chromatography combined with bioassays and other hyphenations—the direct link to the compound indicating the effect, in: B.S. Patil, G.K. Jayaprakasha, F. Pellati (Eds.), Instrumental Methods for the Analysis of Bioactive Molecules, 246th ACS National Meeting and Exposition, September 8–12, Indianapolis, IN, American Chemical Society, Washington, DC, 2013.

Muller MB, Dausend C, Weins C. A new bio-autographic screening method for the detection of estrogenic compounds. Chromatogr. 2004; 60:207-211.

Marton A, Millard M, Hostettmann K. A TLC bioautographic method for the detection of α and β Glucosidase inhibitors in plant extract. GIT Lab. J 1997; 1:36-39.

Dewanjee S, Gangopadhyay M, Bhattacharya N, Khanra R, Dua TK. Bioautography and its scope in the field of natural product chemistry. J Pharma Anal. 2015; 5(2):75-84.

Kraus W, Ngoc LH, Conrad J, Klaiber I, Reeb S, Vogler B. Investigation of biologically active nature products using online LC-bioassay, LC NMR and LC-MC techniques. Phytochem Rev. 2002; 1:409-411.

Cieśla Ł. Biological finger-printing of herbal samples by means of Liquid chromatography. Res Inter. 2012; 26:1-9.

Sticher O. Natural product isolation. Nat Prod Rep. 2008; 25(3):517-554.

Marston A. Thin-layer chromatography with biological detection in phytochemistry. J Chromatogr. 2011; 1218:2676-2683.

Morlock G, Schwack W. Coupling of planar chromatography to mass spectrometry. Trends Anal Chem. 2010; 29(10):1157-1171.

Wagman GH, Bailey JV. Use of silicic acid glass fiber sheets for bioautography of antimicrobial substances. J chromatogr.1969; 41:263-264.

Rios JL, Recio MC, Villar A. Screening methods for natural products with anti microbial activity review of the literature. J Ethnopharmacol. 1988; 23:127-149.

Klingelhöfer I, Morlock GE. Sharp-bounded zones link to the effect in planar chromatography-bioassay-mass spectrometry. J Chromatogr A. 2014; 1360:288-295.

Eberz G, Rast HG, Burger K, Kreiss W, Weisemann C. Bioactivity screening by chromatography-bioluminescence coupling. Chromatogr.

; 43:5-9.

Klöppel A, Grasse W, Brümmer F, Morlock G. HPTLC coupled with bioluminescence and mass spectrometry for bioactivitybased analysis of secondary metabolites in marine sponges. J Planar Chromatogr Modern TLC. 2008; 21(6):431-436.

Krüger S, Urmann O, Morlock GE. Development of a planar chromatographic method for quantitation of anthocyanes in pomace, feed, juice and wine. J Chromatogr A. 2013; 1289: 105-118.

Baumann U, Brunner C, Pletscher E, Tobler N. Biologische Detektionsverfahren in der Dünnschichtchromatographie. Umweltwissenschaften und Schadstoff-Forschung. 2003; 15(3):163-167.

Nagy S, Kocsis B, Kőszegi T, Botz L. Optimization of growth conditions for test fungus cultures used in direct bioautographic TLC detection. 3. Test fungus: Candida albicans. JPC J Planar Chromatogr Modern TLC. 2007; 20(5):385-389.

Schmourl G, Mendonca-Filho RR, Alviano CS. Screening of antifungal agents using ethanol precipitation and bio-autography of medicinal and food plants. J Ethnopharmacol. 2004; 96: 563-568.

Desbrow CE, Routledge EJ, Brighty GC, Sumpter JP, Waldock M. Identification of estrogenic chemicals in STW effluent. 1. Chemical fractionation and in vitro biological screening. Environ Sci Tech. 1998; 32(11):1549-1558.

Aebersold R, Mann M. Mass spectrometry-based proteomics. Nature 2003; 422(6928):198-207.

Riss TE, Moravec R, Niles A. Selecting cell-based assays for drug discovery screening. Cell Notes 2005; 13:16-21.

Hughes JP, Rees S, Kalindjian SB, Philpott KL. Principles of early drug discovery. Br J Pharmacol. 2011; 162(6):1239-1249.

Ideker T, Galitski T, Hood L. A new approach to decoding life: systems biology. Ann Rev Geno Human Gen. 2001; 2(1):343-372.

Endy D. Foundations for engineering biology. Nature 2005; 438:449-453.

Zhang J, Yang PL, Gray NS. Targeting cancer with small molecule kinase inhibitors. Nat Rev Cancer. 2009; 9(1):28-39.

Akkad R, Wolfgang S. Effect of bromine oxidation on highperformance thin-layer chromatography multi-enzyme inhibition assay detection of organophosphates and carbamate insecticides." J Chromatogr A. 2011; 1218(19):2775-2784.

Wen L, Wei Q, Chen G, Liu F, Zhang S, You T. Bioassay and liquid chromatography/mass spectrometry-guided acetylcholinesterase inhibitors from Picria fel-terrae. Pharmacog Mag. 2013; 9(1):S25.

Salazar MO, Furlan RLE A rapid TLC autographic method for the detection of Glucosidase inhibitors. Phytochem Anal. 2007; 18:209-212.

Simoes-Pires CA, Hmicha B, Marston A. A TLC bioautographic method for the detection of α- and β-glucosidase inhibitors in plant extracts. Phytochem Anal. 2009; 20:511-515.

Ramallo IA, Zacchino SA, Furlan RLE. A rapid TLC autographic method for the detection of xanthine oxidase inhibitors and superoxide scavenges. Phytochem Anal. 2006; 17:15-19.

Su X, Li X, Tao H, Zhou J, Wu T, Chou G, Cheng Z. Simultaneous isolation of seven compounds from Glehnia littoralis roots by off‐line over pressured layer chromatography by a TLC antioxidant autographic assay. J Sep Sci.2013; 36(21-22):3644-3650.

Potterat O, Hamburger M. Concepts and technologies for tracking bioactive compounds in natural product extracts: generation of libraries, and hyphenation of analytical processes with bioassays. Nat Prod Rep. 2013; 30(4):546-564.

Pratt DE, Miller EE. A flavonoid anti-oxidant in Spanish peanuts. Am J Oil Chem Soc. 1984; 61:1064-1067.

Colech M, Komsta L, Nowak R Investigation of antiradical activity of plant material by thin layer chromatography with image processing. Food Chem. 2012; 132(1):549-553.

Miller NJ, Rice-Evans CA. Factors influencing the antioxidant activity determined by the ABTS•+ radical cation assay. Free Rad Res. 1997; 26(3):195-199.

Otvos RA, van Nierop P, Niessen WM, Kini RM, Somsen GW, Smit AB, Kool J. Development of an Online Cell-Based Bioactivity Screening Method by Coupling Liquid Chromatography to Flow Cytometry with Parallel Mass Spectrometry. Anal Chem. 2016; 88(9):4825-4832.

Venkateswarlu K, Venisetty RK, Yellu NR, Keshetty S, Pai MG. Development of HPTLC-UV absorption densitometry method for the analysis of alprazolam and sertraline in combination and its application in the evaluation of marketed preparations. J Chromatogr Sci. 2007; 45:537-539.

Alam P. Densitometric HPTLC analysis of 8-gingerol in Zingiber officinale extract and ginger-containing dietary supplements, teas and commercial creams. Asian Pac J Trop Biomed. 2013; 3: 634-638.

Teh SS, Morlock GE.. Analysis of bioactive components of oil seed cakes by high performance Thin-layer chromatographybioassay combined with mass spectrometery. Chromatogr. 2015; 2:125-140.

Cheng Z, Wu T. TLC bioantography: high throughput technique for screening bio-active natural products. Comb. Chem. High Throughput Screen. 2013; 16:531-549.

Verghese S, Narmadha R, Gomathi D, Kalaiselvi M and Devaki K. Phytochemical screening and HPTLC fringer printing analysis of citrullus lanatus (Thunb.) seed. J Acute Dis. 2013; 2:122-126.

Riffault L, Destandau E, Pasquier L, André P, Elfakir C. Phytochemical analysis of Rosa hybrida cv.‘Jardin de Granville’by HPTLC, HPLC-DAD and HPLC-ESI-HRMS: Polyphenolic fingerprints of six plant organs. Phytochem. 2014; 99:127-134.

Das B, Paul T, Apte KG, Chauhan R, Saxen RC. Evaluation of antioxidant potential and quantification of polyphenols of Diplazium esculentum Retz with emphasis on its HPTLC Chromatography. J Pharm Res. 2013; 6:93-100.

Morlock GE. High‐Performance Thin‐Layer Chromatography–Mass Spectrometry for Analysis of Small Molecules. In: Mass spectrometry handbook Lee M (Ed.) John Wiley and Sons: Hoboken, NJ. 2012. 1181-1206p.

Morlock, GE. . Influence of the TLC/HPTLC plate and solvents on the background signals obtained in TLC/HPTLC-ESI-MS using the TLC-MS Interface. J Liq Chromatogr Rel Technol. 2014; 37: 2892–2914.

Weller MG. A unifying review of bio-assay-guides fractionation, effect-directed analysis and related techniques. Sensor. 2012; 12: 9181-9209.

Vogler B, Klaiber I, Roos G, Walter CU, Hiller W, Sandor P, Kraus W. Combination of LC-MS and LC-NMR as a tool for the structure determination of natural products. J Nat Prod. 1998; 61(2):175-178.

Fabel S, Niessner R., Weller MG. Effect-directed analysis by high-performance liquid chromatography with Gas-segmented enzyme inhibition. J Chromatogr. 2005; 1099: 103–110.

Schebb NH, Heus F, Saenger T, Karst U, Irth H, Kool J. Development of a countergradient parking system for gradient liquid chromatography with online biochemical detection of serine protease inhibitors. Anal Chem. 2008; 80: 6764–6772.

Schebb NH, Faber H, Maul R, Heus F, Kool J, Irth H, Karst U. Analysis of glutathione adducts of patulin by means of liquid chromatography (HPLC) with biochemical detection (BCD) and electrospray ionization tandem mass spectrometry (ESIMS/MS). Anal Bioanal Chem. 2009; 394: 1361–1373.

Van Ruth SM. Methods of gas chromatography-olfactometry: A review. Biomol Eng. 2001; 17: 121-128.

Misra K, Tulsawani R, Shyam R, Meena DK, Morlock G, Hyphenated high-performance thin-layer chromatography for profiling of some Indian natural efficiency enhancers. J Liq Chromatogr Rel Tech. 2012; 35(10):1364-1387.

Dytkiewitz E, Morlock GE. Analytical Strategy for Rapid Identification and Quantification of Lubricant Additives Oil by High-Performance Thin-Layer Chromatography with UV Absorption and Fluorescence Detection Combined with Mass Spectrometry and Infrared Spectroscopy. J. AOAC Inter. 2008; 91(5):1237-1244.

Hostettmann K. Strategy for the biological and chemical evaluation of plant extracts. Pure and Applied Chem. 1999; 70:1-8.

Wolfender JL, Ndjoko K, Hostettmann K. Liquid chromatography with ultraviolet absorbance–mass spectrometric detection and with nuclear magnetic resonance spectrometry: a powerful combination for the on-line structural investigation of plant metabolites. J. Chromatogr A. 2003; 1000(1):437-455.

Gössi A, Scherer U, Schlotterbeck G. Thin-layer Chromatography–Nuclear Magnetic Resonance Spectroscopy–A Versatile Tool for Pharmaceutical and Natural Products Analysis. CHIMIA Inter J Chem. 2012; 66(5):347-349.

Accardo G, Cioffi R, Colangelo F, d'Angelo R, De Stefano L, Paglietti F. Diffuse Reflectance Infrared Fourier Transform Spectroscopy for the Determination of Asbestos Species in Bulk Building Materials. Materials. 2014; 7(1):457-470.

Thistlethwaite PJ, Hook MS. Diffuse reflectance Fourier transform infrared study of the adsorption of oleate/oleic acid onto titania. Langmuir. 2000; 16(11):4993-4998.

Somsen GW, Hooijschuur EW, Gooijer C, Brinkman UT, Velthorst NH, Visser T. Coupling of reversed-phase liquid column chromatography and Fourier transform infrared spectrometry using post column on-line extraction and solvent elimination. Analy Chem. 1996; 68(5):746-752.

Rohman A, Man YC. Fourier Transform Infrared (FTIR) spectroscopy for analysis of extra virgin olive oil adulterated with palm oil. Food Res Inter. 2010; 43(3):886-892.

Brosseau CL, Gambardella A, Casadio F, Grzywacz CM, Wouters J, Van Duyne RP. Ad-hoc surface-enhanced Raman spectroscopy methodologies for the detection of artist dyestuffs: thin layer chromatography-surface enhanced Raman spectroscopy and in situ on the fiber analysis. Anal Chem. 2009; 81(8):3056-3062.

Li DW, Zhai WL, Li YT, Long YT. Recent progress in surface enhanced Raman spectroscopy for the detection of environmental pollutants. Microchimica Acta. 2014; 181(1-2):23-43.

Da Silva AP, Rocha R, Silva CM, Mira L, Duarte MF, Florêncio MH. Antioxidants in medicinal plant extracts. A research study of the antioxidant capacity of Crataegus, Hamamelis and Hydrastis. Phytother Res. 2000; 14(8):612-616.

Klejdus B, Mikelová R, Petrlová J, Potešil D, Adam V, Stiborová M, Hodek P, Vacek J, Kizek R, Kubán V. Evaluation of isoflavone aglycone and glycoside distribution in soy plants and soybeans by fast column high-performance liquid chromatography coupled with a diode-array detector. J Agric Food Chem. 2005; 53(15):5848-5852.

Brusotti G, Cesari I, Dentamaro A, Caccialanza G, Massolini G. Isolation and characterization of bioactive compounds from plant resources: The role of analysis in the ethnopharmacological approach. J Pharm Biomed Analy. 2014; 87:218-228.

Nakajima JI, Tanaka I, Seo S, Yamazaki M, Saito K. LC/PDA/ESI-MS profiling and radical scavenging activity of anthocyanins in various berries. BioMed Res Inter. 2004; (5):241-247.

Renukappa T, Roos G, Klaiber I, Vogler B. Kraus W. Application of high performance liquid chromatography coupled to Nuclear magnetic resonance spectrometry, mass spectrometry and bioassay for the determination of active saponins from Bacopa monniera Wettst. J Chromatogr A. 1999; 847:109-116.

Queiroz E.F, Ioset JR, Ndjoko K. Online identification of the bioactive compounds from Blumea gariepina by HPLC-UVMS and HPLC-UV-NMR combined with HPLC microfrationation Phytochem Anal. 2005; 16:166-174.

Rhee IK, Van de Meent M, Ingkaniana K. Screening for acetylcholnesterane inhibitors from Amarylidacea using silica gel thin layer chromatography in combination with bioactivity stainning J Chromatogr A. 2001; 915:217-223.

Zhang H, Shen P, Cheng Y. Identification and determination of the major constituents in traditional Chinese medicine Si-WuTang by HPLC coupled with DAD and ESI–MS. J Pharm Biomed Anal. 2004; 34(3):705-713.

Ibanez E, Kubátová A, Señoráns FJ, Cavero S, Reglero G, Hawthorne SB. Subcritical water extraction of antioxidant compounds from rosemary plants. J Agric Food Chem. 2003; 51(2):375-382.

Hansen SH, Jensen AG, Cornett C, Bjørnsdottir I, Taylor S, Wright B, Wilson ID. High-performance liquid chromatography on-line coupled to high-field NMR and mass spectrometry for structure elucidation of constituents of Hypericum perforatum L. Analy Chem. 1999; 71(22):5235-

Lao SC, Li SP, Kan KK, Li P, Wan JB, Wang YT, Dong TT, Tsim KW. Identification and quantification of 13 components in Angelica sinensis (Danggui) by gas chromatography–mass spectrometry coupled with pressurized liquid extraction. AnalyChem Acta. 2004; 526(2):131-137