Saponins detection

It is marketed as, and has been described to be, a potent adjuvant which has found widespread use in veterinary vaccines against, for example, foot and mouth disease, rabies, and in a number of experimental vaccines and in preclinical trials. Unfortunately its hemolytic activity and local counter reactions make it unsuitable for human vaccines [5]. Furthermore, Quil A is used for production of ISCOMs (immunostimulating complexes, typically composed of 0.5 % Quillaja saponins, 0.1% cholesterol, 0.1% phospholipid and antigen dissolved in PBS). Although side effects of Quil A were almost absent when incorporated into ISCOM, this form of vaccine is only used for veterinary vaccines and has not been approved for humans. Quil A is still a heterogeneous mixture, consisting of up to 23 different individual saponins detectable by HPLC [8]. Later, it was observed that not all saponins were active as adjuvants. A saponin termed QS III was purified from of a methanol extract of Quillaja bark by several chromatographic steps, it has, however, not been tested for adjuvant activity [12]. 

These methods are employed for the detection and determination of antibiotics and substances with similar effects, like alkaloids, insecticides, fungicides, mycotoxins, vitamins, bitter principles and saponins [14]. 

Saponins la 7,411,430 -, bioautographic determination la 109 Sarcosine Ia435 lbl24 Scandium cations, detection la 144 Scanner, optical trains la 30,39 S-Chamber (small chamber) la 126,127 SCHiFF s bases lb 52 Scintillators la 12 Scopolamine lb 231,252,255,323 Scopoletin lb 216-218,365 Screening process lb 45 Sebacic acid la 178,233,249,308 Sebuthylazine lb 418 Selectivity, enhancement by derivatiza-tion la 55... 

The utility of mass selective detection is greatest when analyzing compounds that do not contain chromophores or when structural information is needed for chemical identification. Triterpene saponins contain very weak chromophores and have long been associated with a variety of biological activities including... 

A simple assay for the detection of the malarial parasite Plasmodium falciparum involves saponin lysis of blood sample and membrane filtration followed by amplification of the consensus sequence of eight 21-bp repeats. This procedure has been successfully used in the field (F2). A PCR assay targeting kinetoplast DNA sequences of Leishmania species was also successfully tested in the field (F2). Molecular methods for the detection of toxoplasma gondii and several other parasites have been reviewed (F2). 

Li, L., Tsao, R., Dou, J., Song, F., Liu, Z., and Liu, S. (2005). Detection of saponins in extract of Panax notoginseng by liquid chromatography-electrospray ionisation-mass spectrometry. Anal. Chim. Acta 536,21-28. 

Fig. 3. Immunofluorescence localization of tubulin in microtubules in Swiss 3T3 cultured fibroblasts. Swiss 3T3 cells were fixed in glutaraldehyde followed by treatment with sodium borohydride (4). Tubulin was detected using a rat monoclonal antitubulin antibody (YLl/2), followed by goat antirat IgG conjugated to rhodamine, all steps in the continuous presence of 0.1% saponin. Note the individual microtubules visible under the dark nuclear region (bar = 4 im).

Elias, P. M., Friend, D. S. and Goerke, J. 1979. Membrane sterol heterogeneity. Freeze-fracture detection with saponins and filipin. J. Histochem. Cytochem. 27, 1247-1260. 

Recourse to innovations in NMR spectroscopy is essential for further advances in the investigation of complex saponins. The use of a 600 MHz spectrometer gives the required sensitivity, consistent with the low amounts available by modern isolation techniques, while the use of sequential ID and inverse-detected 2D NMR techniques, couples the short time necessary to perform the experiments with the selfconsistency of the obtained results and, thus, the unambiguous structure assignment [128]. 

CD detection for colored derivatives might just as easily be used for structural information about the carbohydrates as well as for their analysis, but so far little attention has been given to either application. Cyclic oligomers of p-D-glucose have important supporting roles to play in analytical applications that are discussed in a later section. The union of chirality in the carbohydrate moiety of a glycoside metabolite with the unsaturation in the base in such compounds as nucleosides and nucleotides, saponins and flavones, etc., is another area that will ultimately be developed for applications of chiroptical detection methods. New and imaginative ideas are needed for the analysis of carbohydrates, and CD should be one of the favored methods. 

The seeds contain some volatile oil, resin and a large amount of fixed oil (Meisner, 1818). The fruit (without the seeds) contains volatile oil, resin, fat, tannin, pectin and mucilage. The volatile oil (oil of star-anise) amounts to about 4-5% and is almost identical with oil of anise (from P. anisum, LinnS). Star-anise oil from Chinese fruit has a specific gravity at 15°C (59°F) of 0.980-0.990 and its known constituents are anethol, phel-landrene, safrol and hydro-quinone-ethyl-ether (Fliickiger, 1879). Poisonous sikimin has been detected in the fruit (Eykmann, 1881), while Schlegel (1885) found a crystalline principle of a pronounced odour of musk. He also found saponin in the watery extract. 

Publications on the structure and analytical detection of saponins of Panax ginseng and the effect of ginseng on cholesterol synthesis and pyruvate kinase activity in rats have appeared. New saponins include ginsenosides Rhi (46) and... 

The HPLC method has also been described for the determination of astragalosides in "Astragali radix" [59,92,291]. The method was sensitive, rapid and accurate and could be used for quantitative analysis of saponins. Total saponins in extract of plant of Astragalus spp. may be also determined spectrophotometrically after reaction with vanillin in acidic medium by measuring the absorbance at 560 nm (detection range is 40-200 pg) [238,321],... 

Identical sulfated trisaccharide was found in three additional new saponins (2, S, 7). The genin moieties for these saponins correspond to diverse oxidation degrees of saikogenin F. Thus, the epimeric saponins 2 and 5, wth hydroxyl groups at C-29 and C-30, respectively, were also characterised in aerial fractions. A non-sulfated compound 7, holding also a hydroxyl group at C- 29, was detected only in the roots. 

Table 2. Optical rotation, molecular formulae, molecular weight, FAB/MS and sulfate detection of saponins from B, rigidum...

For smaller quantities of compounds more sensitive inverse detected techniques are available, such as HMQC ( IH-I C one bond correlation via heteronuclear multiple quantum coherence, analogous to HETCOR) and HMBC (proton detected heteronuclear multiple bond correlation spectroscopy) (15). The last provide, in addition to the intraresidue multiple bond correlations, interresidue correlations between the anomeric carbon and the aglycone protons.We follow this general strategy for the structural determination of tri terpenoid saponins of Bupleurum fruticosum (16) andArdisia japonica (9). 

Lau, A., Woo, S., and Koh, H.L. 2003. Analysis of saponins in raw and steamed Panax notoginseng using high-performance liquid chromatography with diode array detection. J. Chromatogr. A. 1011, 77-87. 

One of the major problems in nutritional exploitation of tree leaves is the presence of antinutritional and toxic factors (6). The leaves of M. oleifera contain about 1.4% tannins, whereas condensed tannins are not present. Total phenols in leaves ranges from 2.7 - 3.4% (6,100). The M oleifera leaves contain 5.0% saponins as diosgenin equivalent with phytate contents in the leaves of 3. %(6, 100). Activity of trypsin inhibitors and lectins is not detected in the M. oleifera leaves. Other antinntritional factors present in M. oleifera leaves are flatus factors (sucrose + raffinose + stachyose) at 5.6% (107). Nitrate (0.5 tmnol per 100 g) and oxalate (4.1%) are also present inM oleifera leaves (100). The presence of these antinutritional factors in leaves of M. oleifera decreases the bioavailability of other nntrients. However, extraction nsing 80% ethanol decreases the contents of some of these antinntritional factors (6). 

The leaves of M. oleifera have neghgible amounts of tannins and total phenolics (6). Condensed tannins, activity of tiypsin inhibitors and lectins have not been detected in M oleifera. Therefore, these antinutritional factors are considered not to be able to produce any adverse effects in animals. However, lectins have been reported in aqueous extracts at pH 4.5 (43). The saponins are observed to be at comparable contents as in soyabean meal. Phytates are found... 

Finally, since many natural product compounds have been investigated with various chromatographic modes and detection techniques, a selection of examples has been summarized in this chapter. This information has been compiled in the form of tables for well-researched classes of secondary metabolites selected from the major subgroups of isoprenoids (mono-, sesqui-, di-, and triterpenes iridoids and secoiridoids carotenoids saponins and ecdysteroids), of phenolics (coumarins, flavonoids, and isoflavonoids), and of alkaloids. 

R76 N. P. Sahu and B. Achari, Advances in Structural Determination of Saponins and Terpenoid Glycosides , p. 315 R77 M. Liu and J. C. Lindon, Recent Advances in Editing and Selective Detection Methods for NMR Spectroscopy , p. 351... 

The following method (Bohn, 1978, 1980) is particularly suitable for monolayer cells. Fixation with FA (1%) and 0.0125% GA gives poor penetration of sera, even after 18 h of incubation. Saponin enhances penetration, and thus the detection of intracellular antigens whereas pretreatment with saponin is unsatisfactory for the preservation of morphology. Several combinations of GA and saponin were investigated at constant FA concentrations. Best results were obtained with the mixture given in Table 17.9D and Table 17.10. 

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