Yanhua Gao

Comparative Study on Separation and Purification of Isoflavones from the Seeds and Sprouts of Chickpea by High-Speed Countercurrent Chromatography

Abstract Chickpea is known as a plant that is rich in protein, carbohydrates, and nutrition, and its seeds and sprouts have been processed into various health foods. In the present study, four isoflavones were purified from the seeds and sprouts of chickpea by high-speed countercurrent chromatography (HSCCC) using two biphasic solvent systems composed of n-hexane–ethyl acetate–methanol–water (5:5:5:5, v/v) and ethyl acetate–water (1:1, v/v). The results indicated that 14.2 mg of formononetin, 15.7 mg of biochanin A, 9.1 mg of ononin, 11.3 mg of biochanin A-7-O-β-D-glucoside were obtained from 150 mg of sprout extracts with the purity of 92.26%, 95.86%, 95.32%, and 96.56%, respectively. Compared with the sprouts, separation of seed extracts yielded less amounts of biochanin A-7-O-β-D-glucoside and biochanin A with lower purity. The results indicate that four main isoflavones in chickpea, i.e., isoflavones, formononetin, biochanin A, ononin, and biochanin A-7-O-β-D-glucoside, are substantially increased by biosynthesis during the seed germination.

Isoflavones extracted from chickpea Cicer arietinum L. sprouts induce mitochondria-dependent apoptosis in human breast cancer cells.

Isoflavones are important chemical components of the seeds and sprouts of chickpeas. We systematically investigated the effects of isoflavones extracted from chickpea sprouts (ICS) on the human breast cancer cell lines SKBr3 and Michigan Cancer Foundation‐7 (MCF‐7). 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assays showed that ICS (10–60 µg/mL) significantly inhibited the proliferation of both cell lines in a time‐dependent and dose‐dependent fashion. Wright‐Giemsa staining as well as annexin V‐fluorescein isothiocyanate and propidium iodide (Annexin V/PI) staining showed that ICS significantly increased cytoclasis and apoptotic body formation. Quantitative Annexin V/PI assays further showed that the number of apoptotic cells increased in a dose‐dependent manner following ICS treatment. Semiquantitative reverse transcription PCR showed that ICS increased the expression of the apoptosis‐promoting gene Bcl‐2‐associated X protein and decreased the expression of the antiapoptotic gene Bcl‐2. Western blot analysis showed that treatment of SKBr3 and MCF‐7 cells with ICS increased the expression of caspase 7, caspase 9, P53, and P21 in a dose‐dependent manner. Flow cytometry assays using the fluorescent probe 3,3′‐dihexyloxacarbocyanine iodide showed a dose‐dependent decrease in mitochondrial membrane potential following ICS treatment. Treatment using ICS also induced a dose‐dependent increase in reactive oxygen species production. This is the first study to demonstrate that ICS may be a chemopreventive or therapeutic agent against breast cancer. Copyright

Isolation of Antimicrobial Peptides from Fritillaria pallidiflora

Polypeptide fractions from seeds of Fritillaria pallidiflora Schrenk were isolated and purified. The peptide compositions and molecular masses were determined using SDS-PAAG electrophoresis and LC-MS analysis. The results showed that the most active fraction of biocidal peptides contained greater than 20 constituents with molecular masses in the range of 1050–4727.2 Da. The isolated peptide fractions exhibited antimicrobial activity against the fungus Candida albicans.

Isolation of two antioxidant peptides from seeds of Apium graveolens indigenous to Chinaa

China National Funds for Distinguished Young Scientists 30925045; CAS/SAFEA International Partnership Program for Creative Research Teams 2008-18

Isolation, Characterization, and Antimicrobial Activity of Extracellular Polypeptides from Diazotrophic Strain Rhizobium radiobacter

Antimicrobial polypeptide fractions were isolated from Rhizobium radiobacter culture medium and purified. Mass-spectrometric analysis showed that the most active biocidal peptide fraction contained 24 components with molecular masses in the range 1171.14–3626.78 Da. The isolated peptide fractions contained 60 and 56% protein, respectively, and inhibited the growth of test pathogens Candida albicans (CA) and Escherichia coli (EC). These polypeptides demonstrated protective properties.

POLYSACCHARIDES FROM Ferula sinkiangensis AND POTENT INHIBITION OF PROTEIN TYROSINE PHOSPHATASE 1B

Ferula sinkiangensis is known as a medicinal plant for thousands of years in China. It is a perennial plant species endemic in Xinjiang and is used in traditional Uyghur medicine for a long time 1 . There are many reports on the pharmacological activities of Ferula, which include treating dysentery, counteracting toxins killing parasitic worms, and resolving phlegm. It is also used as a deodorant. Previous investigations of the chemical composition of Ferula have identified essential oils, flavonoids, minerals, and ferulic acid in the extract [2]. Most reports of Ferula have concentrated on the pharmacological aspects of these compounds. Only a few reports have described methods for the extraction of the polysaccharides, and none has described the isolation or identification of these compounds. To the best of our knowledge, there is no report about polysaccharides from roots of Ferula.This study will provide a basis for future research and application of these polysaccharides. The goal of the current work was to establish a method to isolate and quantitate the polysaccharides from roots of Ferula, as well as to determine their monosaccharide compositions, elucidate their structures, and evaluate water-soluble polysaccharides (FSPs), a potent inhibitor of protein tyrosine phosphatase 1B (PTP1B) in vitro. Lipids, pigments, and low-molecular-weight compounds were removed from air-dried raw material by extraction with petroleum ether, CHCl3, and MeOH [3]. The remaining raw material was dried and then extracted with hot water to obtain FSPs. The yield was 9.5%. The total sugar and protein were measured by anthrone-sulfuric acid and Bradford methods [4, 5] with D-glucose and bovine serum albumin (BSA) as standard. The analysis of total sugar and protein contents indicated that FSPs was composed of 45.1 3% polysaccharide and 12.5 3% protein. Furthermore, uronic acid measurement by the m-hydroxydiphenol method [6] with D-galacturonic acid as standard indicated that 28.1 3% of the polysaccharide moiety in FSPs was acidic. Consequently, the preliminary results demonstrated that FSPs was an acid proteoglycan. After removing proteins by the Savage method [7], the crude polysaccharides (FSPs) gave a negative response to the Bradford test, and no absorption was detected at 280 and 260 nm, indicating the absence of protein and nucleic acids. The neutral polysaccharide, eluted with distilled water, was named FSPs-n. The acidic polysaccharides, eluted by the stepwise addition of 0–1.0 M NaCl solutions, was named FSPs-a. The yields of the two fractions were 6.8 and 14.8% in the crude polysaccharides [8, 9]. According to the results of HPLC analysis [10, 11], the carbohydrate portion of FSPs contained ribose, arabinose, glucose, fucose, and galactose in a ratio of 8.9:3.3:2.1:1.5:0.3. FSPs-n was composed of glucose, xylose, arabinose, galactose, and mannose in a ratio of 3.9:4.0:1.8:1.4:0.8, and FSPs-a contained glucose, xylose, mannose, and arabinose in a ratio of 6.5:4.0:1.7:1.0. Furthermore, FSPs and FSPs-a were composed of glucuronic acid and galacturonic acid in a ratio of 1.1:0.3 and 2.3:5.5, respectively.

A new isoflavane glycoside from Cicer arietinum seeds

A new isoflavane glycoside with a novel structure that we called cicerarietinuoside A (1) in addition to the three known compounds β-daucosterol (2), adenosine (3), and ent-kaurane diterpene glycoside (4) were isolated from seeds of chickpea. Their structures were established based on broad spectral analysis. Compounds 2 and 4 were obtained from chickpea seeds for the first time.