Abstract
Saponins are commonly found in adequate amounts in the root tissue of plant, however recent studies have reported that saponins can be also found in considerable amounts in plant aerial tissues such as leaf and stem. Thus, quantification of total saponin contents in different plant species and organs are very important to understand their biological functions in plant defense. There are several methods have been developed for measuring saponin contents in medicinal as well as crop plant species. The classical colorimetric and biological methods are remain popular methods for saponin quantification. However, biological and colorimetric determinations of saponin contents doesn’t provide accurate information and sometimes might resulted in a misleading information, due the large structural variation of individual saponins not only within different species, but even also among same species. Thus, more sensitive methods have been recently introduced to measure and quantify saponin contents in different plant extracts. High performance (HP)-thin-layer chromatography (TLC) on normal (HPTLC) or reversed-phase (two-dimension, 2D-HPTLC) provides more precise and reliable saponin qualitative information, especially when these HPTLC methods are combined with a computer flying-spot scanner with dual-wavelength. After screening the saponin profile on the TLC, a 2D-analytical software can applied for the quantification of saponin level in plant extracts. However, for reliable measurements a proper saponin standards must be run with the saponin extracts for comparative analysis. Standardization and identification of the peaks by HPLC chromatograms has been also developed for saponin quantification, which relay on the comparisons of the retention times with those observed for authentic standards. On the other hand, there are limited applications of gas chromatography (GC) for quantification and determination of saponin compounds, due to the high molecular weights of the saponin compounds. In this chapter we will discussed some of these methods and the amount of saponin detected in different plant species.
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References
Abdelrahman M, Hirata S, Ito SI, Yamauchi N, Shigyo M (2014) Compartmentation and localization of bioactive metabolites in different organs of Allium roylei. Biosci Biotechnol Biochem 78:1112–1122
Abdelrahman M et al (2017) RNA-sequencing-based transcriptome and biochemical analyses of steroidal saponin pathway in a complete set of Allium fistulosum—A. cepa monosomic addition lines. PLoS One 12:e0181784
Berhow MA, Cantrell CL, Duval SM, Dobbins TA, Mavnes J, Vaughn SF (2002) Analysis and quantitative determination of group B saponins in processed soybean product. Phytochem Anal 13:343–348
Burnouf-Radosevich M, Delfel NE (1986) High-performance liquid chromatography of triterpene saponins. J Chromatogr 368:433–438
Bushway RJ, Barden ES, Bushway AW, Bushway AA (1979) High-performance liquid chromatographic separation of potato glycoalkaloids. J Chromatogr A 178:533–541
Chaicharoenpong C, Petsom A (2009) Quantitative thin layer chromatographic analysis of the saponins in tea seed meal. Phytochem Anal 20:253–255
Coran SA, Mulas S (2012) Validated determination of primulasaponins in primula root by a high-performance-thin-layer-chromatography densitometric approach. J Pharm Biomed Anal 70:647–645
Coulson CB (1958) Saponins. I.-Triterpenoid saponins from lucerne and other species. J Sci Food Agric 9:281–288
Edewor TI, Owa SO, Ologan AO, Akinfemi F (2016) Quantitative determination of the saponin content and GC-MS study of the medicinal plant Cassytha fiiformis (linn.) leaves. J Coastal Life Med 4:154–156
Gnoatto SCB, Schenkel EP, Bassani VL (2005) HPLC method to assay total saponins in Ilex paraguariensis aqueous extract. J Braz Chem Soc 16:1678–4790
Gorski PM, Jaworski A, Shannon S, Robinson RW (1985) Rapid TLC and KPLC test for cucurbitacins. Genet Coop Rep 8:69–70
Gorski PM, Jaworski A, Shannon S, Robinson RW (1986) Rapid TLC and HPLC quantification of cucurbitacin C in cucumber cotyledons. HortScience 21:1034–1036
Gu L, Tao G, Gu W, Prior RL (2002) Determination of soyasaponins in soy with LC-MS following structural unification by partial alkaline degradation. J Agric Food Chem 50:6951–6959
Gurfinkel DM, Rao AV (2002) Determination of saponins in legumes by direct densitometry. J Agric Food Chem 50:426–430
James J, Dubery I (2011) Identification and quantification of triterpenoid centelloids in Centella asiatica (L.) urban by densitometric TLC. J Planar Chromatogr Modern TLC 24:82–87
Kasai R, Yamaguchi H, Tanaka O (1987) High-performance liquid chromatography of glycosides on a new type of hydroxyapatite column. J Chromatogr A 407:205–210
Kawahara Y, Hoshino T, Morimoto H, Shinizu T, Narukawa Y, Fuchino H, Kawahara N, Kiuchi F (2016) LC-MS-based quantification method for Achyranthes root saponins. J Nat Med 70:102–106
Kesselmeier J, Strack D (1981) High performance liquid chromatography analysis of steroidal saponins from Avena sativa L. Zeitschrift fur Naturforschung Sect C J Biosci 36:1072–1074
Khakimov B, Motawia MS, Bak S, Engelsen SB (2013) The use of trimethylsilyl cyanide derivatization for robust and broad-spectrum high-throughput gas chromatography-mass spectrometry based metabolomics. Anal Bioanal Chem 405:9193–9205
Khalil AH, El-Adawy TA (1994) Isolation, identification and toxicity of saponin from different legumes. Food Chem 50:197–120
Kim Y, Wampler DJ (2009) Determination of saponin and various chemical compounds in Camellia sinensis and genus Ilex. Sensus Tech Note (SEN-TN-0027)
Kimata H, Sumida N, Matsufuji N, Morita T, Ito K, Yata N, Tanaka O (1985) Interaction of saponin of bupleuri radix with ginseng saponin: solubilization of saikosaponin-a with chikusetsusaponin V (= ginsenoside-Ro). Chem Pharm Bull (Tokyo) 33:2849–2853
Kite GC, Porter EA, Simmonds MS (2007) Chromatographic behaviour of steroidal saponins studied by high-performance liquid chromatography-mass spectrometry. J. Chromatogr A 1148:177–183
Li XE, Wang YX, Sun P, Liao DO (2016) Determination of saponin content in Hang Maidong and Chuan Maidong via HPLC-ELSD analysis. J Anal Meth Chem 2016:7214607
Lin J, Wang C (2006) An analytical method for soy saponins by HPLC/ELSD. Food Sci 69:C456–C462
Liu R, Cai Z, Xu B (2017) Characterization and quantification of flavonoids and saponins in adzuki bean (Vigna angularis L.) by HPLC–DAD–ESI–MS analysis. Chem Central J 11:93
Liu Y-M, Zhou A, Yu N-J, Han R-C, Zhang W, Zhu Y-J, Cao Y, Li X-Y, Peng D-Y (2018) Simultaneous determination of five saponins in Bupleuri Radix by HPLC-DAD dual wavelength method. Zhongguo Zhong Yao Za Zhi 43:363–368
Mackie AM, Singh HT, Owen JM (1977) Studies on the distribution, biosynthesis and function of steroidal saponins in echinoderms. Comp Biochem Physiol Part B Comp Biochem 56:9–14
Mostafa A et al (2013) Aginoside saponin, a potent antifungal compound, and secondary metabolite analyses from Allium nigrum L. Phytochem Lett 6:274–280
Muir AD, Ballantyne KD, Hall TW (2000) LC-MS and LC-MS/MS analysis of saponins and sapogenins—comparison of ionization techniques and their usefulness in compound identification. In: Oleszek W, Marston A (eds) Saponins in food, feedstuffs and medicinal plants. Proceedings of the Phytochemical Society of Europe, vol 45. Springer, Dordrecht
Naidu MM, Shyamala BN, Naik JP, Sulochanamma G, Srinivas P (2011) Chemical composition and antioxidant activity of the husk and endosperm of fenugreek seeds. LWT Food Sci Technol 44:451–456
Nascimento YM, Abreu LS, Lima RL, Costa VCO, de Melo JIM, Braz-Filho R, Silva MS, Tavares JF (2019) Rapid characterization of triterpene saponins from Zornia brasiliensis by HPLC-ESI-MS/MS. Molecules 24:2519
Negi JS, Singh P, Pant GJN, Rawat MSM (2011) High-performance liquid chromatography analysis of plant saponins: An update 2005-2010. Pharmacogn Rev 5:155–158
Nowacka J, Oleszek W (1997) High performance liquid chromatography of zanhic acid glycoside in alfalfa (Medicago sativa). Phytochem Analysis 3:227–230
Nyakudya E, Jeong JH, Lee NK, Jeong Y-S (2014) Platycosides from the roots of Platycodon grandiflorum and their health benefits. Prev Nutr Food Sci 19:59–68
Oleszek WA (2002) Chromatographic determination of plant saponins. J Chromatogr A 967:147–162
Peng Y, Luo F, Wang S, Li L, Sun Y, Pan W (2008) Determination of sarsasapogenin in Rhizoma Anemarrhenae with precolumn derivatization by HPLC. J Shenyng Pharm Univ 25:372–375
Podolak I, Hubicka U, Zuromska-Witek B, Janeczko Z, Krzek J (2013) Quantification of saponins in different plant parts of Lysimachia L. species by validated HPTLC-densitometric method. J Planar Chromatogr Modern TLC 26(3)
Saito K, Horie M, Hoshino Y, Nose N, Nakazawa H (1990) High-performance liquid chromatographic determination of glycoalkaloids in potato products. J Chromatogr 508:141–147
Shawky E, Sallam SM (2017) Simultaneous determination of soyasaponins and isoflavones in soy (Glycine max L.) products by HPTLC-densitometry-multiple detection. J Chrom Sci 55:1059–1065
Slacanin I, Marston A, Hostettmann K (1988) High-performance liquid chromatographic determination of molluscicidal saponins from Phytolacca dodecandra (Phytolaccaceae). J Chromatogr 448:265–274
Soni N, Singh VK, Singh DK (2020) HPLC characterization of molluscicidal component of Tamarindus indica and its mode of action on nervous tissue of Lymnaea acuminate. J. Ayurveda Integr Med 11:131–139
Tagiev SA, Ismailov AI (1986) Quantitative determination of gyposoide in roots of Gypsophilla bicolor Grossh. Rastit Resur 22:262–265
Tal DM, Patrick PH, Elliott W (1984) Bile acids. Lxx. preparative separation of kryptogenin from companion sapogenins by high performance liquid chromatography. J Liquid Chromatogr 7:2591–2603
Tie-xin T, Hong W (2008) An image analysis system for thin-layer chromatography quantification and its validation. J Chromatogr Sci 46:560–564
Uematsu Y, Hirata K, Saito K (2000) Spectrophotometric determination of saponin in Yucca extract used as food additive. J AOAC Int 83(6)
Van Atta GR, Guggolz J, Thompson CR (1961) Plant analysis, determination of saponins in alfalfa. J Agric Food Chem 9:77–79
Wagner H, Bladt S, Zgainski EM (1986) Plant Drug Analysis-a thin layer chromatography atlas. Springer, Berlin
Wei F, Ma L-Y, Cheng X-L, Lin R-C, Jin W-T, Khan IA, Lu JQ (2005) Preparative HPLC for Purification of Four Isomeric Bioactive Saponins from the Seeds of Aesculus chinensis. J Liquid Chromatogr Related Technol 28(5)
Xu C-J, Lin J-T (1985) Comparison of silica-, C18-, and NH2-Hplc columns for the separation of neutral steroid saponins from dioscorea plants. J Liquid Chromatogr 8:361–368
Zhang Q (1995) Determination of oleanolic acid in roots of Achyranthes bidentata by TLC-scan. Chin Pharm J 30:592–594
Zhang GD, Zhou ZH, Liu HY (1983) Analysis of ginseng. III. Isolation and determination of ginseng saponins. Acta Pharmaceutica Sinica 18:607–611
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Abdelrahman, M., Jogaiah, S. (2020). Method of Estimation in Biological Sample. In: Bioactive Molecules in Plant Defense. Springer, Cham. https://doi.org/10.1007/978-3-030-61149-1_7
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DOI: https://doi.org/10.1007/978-3-030-61149-1_7
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