A scaled-up SRCCC apparatus equipped with a 40-L column was constructed on the basis of separations of crude broccoli seed extract and crude radish seed extract using a conventional SRCCC instrument. Scaled-up separation of 500 g of crude broccoli seed extract with n-butanol-acetonitrile-10% (NH4)2SO4 aqueous solution 1:0.5:2 (υ/υ) as solvent system yielded 61.5 g glucoraphanin product of purity 91.2%. Separation of 500 g crude radish seed extract with the same solvents in the ratio 0.5:1:2 (υ/υ) afforded 26.7 g glucoraphenin product of purity 94.5%. Recovery of glucoraphanin and glucoraphenin from the crude extracts was 98.3 and 98.9%, respectively. The results demonstrated that this SRCCC technology is useful for separation of glucosinolates from plant extracts.
Authors:M.Y. Jiang, Z.R. Wang, K.W. Chen, J.Q. Kan, K.T. Wang, Zs. Zalán, F. Hegyi, K. Takács, and M.Y. Du
After suffering from mechanical injury and fungal infection, grapes are perishable. Botrytis cinerea, the causal agent of gray mould, is a critical pathogen for grapes. In this study, the inhibitory effect of Pseudomonas fluorescens on the formation of gray mould on grapes during the postharvest storage was investigated on “Kyoho” grape. The results suggest that a living cell suspension of P. fluorescens significantly inhibited spore germination of B. cinerea, and significantly reduced the incidence of grape gray mould. Moreover, compared with the control, the fruit inoculated with P. fluorescens had elevated activities of polyphenol oxidase (PPO), peroxidase (POD), catalase (CAT), phenylalanine ammonia-lyase (PAL), chitinase (CHI), and β-1,3-glucanase (GLU). Increase in enzyme activity correlated with enhanced host resistance. In addition, there was little difference in storage quality between the treated group and control group, indicating no adverse effects of the induced defence response on fruit quality.
Authors:Q. Du, L. Zhang, H. Cui, K. Wang, L. Xiu, and L. Sheng
Cleavage of glucosinolates with myrosinase yields thioglycosidic compounds which have cancer chemoprevention activity. In this paper, glucosinolates in an extract (2.0 g) of broccoli seeds (Brassica oleracea var. italica) were separated by high-speed countercurrent chromatography (HSCCC) with the solvent system n-butanol-acetonitrile-10% ammonium sulfate solution 1:0.5:2.2 (v/v) to yield five glucosinolate compounds after desalting and decolorizing by MCI column chromatography. The five compounds, 7-methylsulfinylheptyl glucosinolate (22.4 mg), 4-pentenyl glucosinolate (33.6 mg), 3-butenyl glucosinolate (24.0 mg), 4-methylsulfinylbutyl glucosinolate (161.4 mg), and 3-methylsulfinylpropyl glucosinolate (29.6 mg), were identified by ESI-MS, 1H NMR and 13C NMR. The purity of the products was >98%, and 7-methylsulfinylheptyl glucosinolate was obtained from broccoli seeds for the first time.
Authors:T.-K. Wang, Y.-D. Yang, B. Du, S. Yu, and W.-L. Hou
Chestnut exhibits anti-inflammatory, styptic, anti-diarrhea, and analgestic effects as a traditional Chinese medicine. There is increasing evidence that shows that the consumption of chestnuts has become more important in human nutrition due to the health benefits provided by the antioxidants. The phenolic compounds are responsible for major bioactivities, such as anti-tumor and anti-oxidation. A high-performance liquid chromatography (HPLC) method with diode array detection (DAD) was established for the simultaneous determination of six phenolic compounds (gallic acid, GA; protocatechuic acid, PR; catechin, CA; epicatechin, EP; quercetin, QU; kaempferol, KA) in Chinese chestnut (Castanea mollissima blume) kernel. The sample followed by separation on Eclipse XDB-C18 column (150 × 4.6 mm, id., 5 μm) with gradient elution of methanol-1.0% acetate acid solution as a mobile phase, at a temperature of 30°C, under the ratio of 1.2 mL min−1, with 5 μL injection volume, and multi-wavelength synthesis was used with DAD. The correlation coefficients were larger than 0.999, the recoveries were 97.58% for GA, 100.41% for PA, 96.23% for CA, 101.38% for QU, 99.15% for EP, and 98.60% for KA, relative standard deviation (RSD) were 1.04% for GA, 1.21% for PA, 1.09% for CA, 1.19% for QU, 1.06% for EP, and 1.20% for KA. This method was applied for the determination of phenolics in chestnut kernel and was found to be fast, sensitive, and suitable.
Authors:Y. C. Xiao, L. T. Liu, J. J. Bian, C. Q. Yan, L. Ye, M. X. Zhao, Q. S. Huang, W. Wang, K. Liang, Z. F. Shi, and X. Ke
Shuganjieyu (SGJY) capsule is a classical formula widely used in Chinese clinical application. In this paper, an ultra-performance liquid chromatography coupled with electrospray ionization and ion trap mass spectrometry has been established to separate and identify the chemical constituents of SGJY and the multiple constituents of SGJY in rats. The chromatographic separation was performed on a C18 RRHD column (150 × 2.1 mm, 1.8 μm), while 0.1% formic acid–water and 0.1% formic acid–acetonitrile was used as mobile phase. Mass spectral data were acquired in both positive and negative modes. On the basis of the characteristic retention time (Rt) and mass spectral data with those of reference standards and relevant references, 73 constituents from the SGJY and 15 ingredients including 10 original constituents and 5 metabolites from the rat plasma after oral administration of SGJY were identified or tentatively characterized. This study provided helpful chemical information for further pharmacology and active mechanism research on SGJY.
Authors:E. Horvath-Szanics, J. Perjéssy, A. Klupács, K. Takács, A. Nagy, E. Koppány-Szabó, F. Hegyi, E. Németh-Szerdahelyi, M.Y. Du, Z.R. Wang, J.Q. Kan, and Zs. Zalán
The increasing consumer demand for less processed and more natural food products – while improving those products’ quality, safety, and shelf-life – has raised the necessity of chemical preservative replacement. Biopreservation refers to extended storage life and enhanced safety of foods using the natural microflora and (or) their antibacterial products. Chitinolytic enzymes are of biotechnological interest, since their substrate, chitin, is a major structural component of the cell wall of fungi, which are the main cause of the spoilage of food and raw plant material. Among the several organisms, many bacteria produce chitinolytic enzymes, however, this behaviour is not general. The chitinase activity of the lactic acid bacteria is scarcely known and studied.
The aim of the present study was to select Lactobacillus strains that have genes encoding chitinase, furthermore, to detect expressed enzymes and to characterise their chitinase activity. Taking into consideration the importance of chitin-bindig proteins (CBPs) in the chitinase activity, CBPs were also examined. Five Lactobacillus strains out of 43 strains from 12 different species were selected by their chitinase coding gene. The presence of the chitinase and chitin-biding protein production were confirmed, however, no chitinolytic activity has been identified.