Four cyclic peptides were isolated from the 75% ethanol extract of the fibrous roots of Pseudostellaria heterophylla by silica gel, Sephadex LH-20 column chromatography, and semi-preparative HPLC. Through mass spectrometry, NMR and other methods, they were identified as pseudostellarin L(1), heterophyllin B(2), pseudostellarin B(3), and pseudostellarin C(4). Among them, compound 1 was a new cyclic peptide, and compounds 2-4 were isolated from the fibrous roots of P. heterophylla for the first time. None of these compounds displayed cytotoxic activities against MCF-7, A549, HCT-116, and SGC-7901 cells.
Caryophyllaceae/chemistry* ; Chromatography, High Pressure Liquid ; Magnetic Resonance Spectroscopy ; Peptides, Cyclic/pharmacology* ; Plant Roots/chemistry*

Referring to the rules for agricultural seed testing (GB /T 3543-1995) issued by China, the test of sampling, seed purity, weight per 1 000 seeds, seed moisture, seed viability and germination rate had been studied for screening seed quality test methods of Pesudostellaria heterophylla. The seed quality from different collection areas was measured. The results showed that at least 6.5 g seeds should be sampled and passed through 10-mesh sieve for purity analysis. The weight of 1 000 seeds was determined by using the 500-seed method. The phenotypic observation and size measurement were used for authenticity testing. The seed moisture was determined under the higher temperature (130 ± 2) degrees C for 5 hours. The seeds were dipped into 0.2% TTC sustaining 1 hour at 40 degrees C, then the viability could be determined. The break dormancy seeds were cultured on sand at 10 degrees C. K cluster analysis was applied for the data analysis, the seed quality from different collection areas grading of P. Heterophylla was described as three grades. The seed quality of each grade should reach following requirements: for first grade seeds, germination rate ≥ 86%, 1 000-grain weight ≥ 2.59 g, purity ≥ 87%, moisture ≤ 13.1%; for second grade seeds, germination rate ≥ 70%, 1 000-grain weight ≥ 2.40 g, purity ≥ 77%, moisture ≤ 14.3%; for third grade seeds, germination rate ≥ 41%, 1 000-grain weight ≥ 2.29 g, purity ≥ 76%, moisture ≤ 15.8%. The seed testing methods for quality items of P. heterophylla had been initially established, as well as the primary P. heterophylla seed quality classification standard.
Caryophyllaceae ; chemistry ; growth & development ; Germination ; Quality Control ; Seeds ; chemistry ; growth & development ; Water ; analysis

OBJECTIVETo study the commodity specification and grade standard of Pseudostellaria Radix, for standardizing market order and achieving industrialization, standardization and modernization of Pseudostellariae Radix.

METHODThe different areas and grade medicinal materials of Pseudostellariae Radix were respectively measured in quantitative characteristics of appearance, and the present grade classification of Pseudostellaria Radix was scientifically verified by the content of polysaccharide. Then the determination data were evaluated by spss correlation analysis, principal component analysis and cluster analysis. So combining with the actual production, the commodity grade standard of Pseudostellariae Radix was formulated.

RESULTCorrelation analysis indicated that the present grade classification of Pseudostellaria Radix was reasonable, and the more the grade of Pseudostellariae Radix was high, the more the content of polysaccharide was high. Meanwhile, length as a classification index was not suitable for the commodity grade standard of Pseudostellariae Radix. Using principal component analysis and cluster analysis, combining actual production, the thickest diameter, weight of single root tuber and the number of 50 g root tuber were filtrated and the grade was divided into 5 ranks: big, mid- dle, small selected goods, big ungraded goods and small ungraded goods.

CONCLUSIONthe commodity specification and grade standard of Pseudostellariae Radix that mainly included the thickest diameter, weight of single root tuber and the number of 50 g root tuber was formulated, the standard was divided into 5 grade. Each grade was not only consistent with the present situation of medicinal materials market, it could also reflected the intrinsic quality of Pseudostellariae Radix. In conclusion, the standard could be used as a classifica- tion basis to the commodity specification and grade.

Caryophyllaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; standards ; Quality Control ; Reference Standards

OBJECTIVETo provide new germplasm materials for breeding new varieties of Pseudostellaria heterophylla.

METHODThe method of single plant selection was adopted, with the comparative experiments being carried out under the same conditions in Shibing county. The 8 plants of Shibing SB-4 were compared respectively with factor analysis for 27 phenotypic traits and 8 yield traits, and single factor variance analysis for the contents of polysaccharides.

RESULTUsing factor analysis, 27 phenotypic traits were classified into 7 principal divisors and 8 yield traits were simplified into 3 principal divisors. The 4 strains of P. heterophylla, ZT-01, ZT-02, ZT-06 and ZT-07, performed better than others in the phenotypic traits, and ZT-01, ZT-02, ZT-03 and ZT-07 in the yield traits. The contents of polysaccharides of ZT-01, ZT-02, ZT-05 and ZT-08 showed significantly higher value.

CONCLUSIONThere is significant difference among the 8 strains of P. heterophylla in phenotypic traits, yield traits and quality traits, making it possible to select certain strains for different purposes. ZT-01 and ZT-02 can be breaded further. ZT-06 and ZT-07 were used as ornamental cultivars for its great phenotypic traits. ZT-03 with good resistance and high yield was taken as resistant variety, and ZT-05 would face next selection on the basis of its high content of polysaccharide.

Breeding ; Caryophyllaceae ; chemistry ; growth & development ; China ; Phenotype ; Polysaccharides ; analysis

A method for determintion of allantion in Psammosilene tunicoides was established by HPLC. Using alcohol as the extraction solvent, the subsequent filtrate of P. tunicoides was analysed by HPLC. Allantoin was successfully detected and separated by ZORBAX NH2 column (4.6 mm x 150 mm,5 microm) at wavelength of 220 nm and column temperature of 40 degrees C, with acetonitrile-water (93: 7) as mobile phase at a flow rate of 1.0 mL x min(-1). The results showed that it had a good linear relationship between the concent ration of allantion and chromatographic peak area. The linear correlation coefficient of allantion was 0.999 5 in 0.010 4-0.166 g x L(-1). The relative standard deviation of six parallel injections was less than 2.1%. The average recoveries were ranged from 95.47% to 100.9%. This method was sensitive and accurate for the determination of allantion in P. tunicoides.
Allantoin ; analysis ; Animals ; Caryophyllaceae ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Stability ; Humans ; Rats ; Reproducibility of Results

OBJECTIVETo develop an effective identification method for accurately discriminating Psammosilene tunicoides and its confused species by the combined method of microscopic identification and molecular identification, so-called systematic identification of Chinese materia medica (SICMM).

METHODP. tunicoides and its confused species were accurately discriminated by SICMM method, which was established by comprehensively use of microscopic identification and DNA identification method. The DNA identification included the following analysis: the BLAST alignment, specific bases and N-J phylogenetic tree analysis.

RESULTThe cluster crystals were not observed in P. tunicoides, but great deals of them were found in Silene viscidula. Further more, big differences of ITS sequence were observed and analyzed between P. tunicoides and its confused specie of S. viscidula.

CONCLUSIONThe system method is a scientific and accurate method for the identification of P. tunicoides and its counterfeit species.

Base Sequence ; Caryophyllaceae ; chemistry ; classification ; cytology ; genetics ; DNA, Intergenic ; Phenotype ; Phylogeny ; Sequence Alignment

Three compounds, including a new one (1), were isolated from the roots of Psammosilene tunicoides. Based on analysis of NMR data as well as their physical and chemical properties, their structures were elucidated as 3,3'-(4,5-dimethoxynaphthalene-2,7-diyl) bis (1-nitropropan-1-one) (1), alpha-spinasterol-3-O-beta-D-glucopyranoside (2) and alpha-spinasteryl-3-O-beta-D-glucoside-6'-O-palmitate(3). Compounds 2 and 3 were isolated from this genus for the first time.
Caryophyllaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Roots ; chemistry ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo study the chemical constituents and bacteriostatic activity of Psammosilene tunicoides.

METHODSuch methods as silica gel column chromatography and gel column chromatography were adotped separate and purify the compounds, and their structures were indentified on the basis of their spectral data and physicochemical properties.

RESULTTen compounds were separated from ethanol extracts and identified as methyl-4-hydroxybenzoate (1), N-methylsaccharin (2), 3-hydroxy-4-methoxybenzoic acid (3), germanicol (4), tricosanoic acid (5), octacosane (6), amber acid (7), succinic acid (8), stellarine A (9), and oleanane-12-ene-3alpha, 16alpha-two hydroxy-23,28-acid (10).

CONCLUSIONAll compounds except compound 10 were separated from P. tunicoides for the first time. Compounds 8 and 9 showed the bacteriostatic activity to a certain extent.

Anti-Bacterial Agents ; chemistry ; isolation & purification ; pharmacology ; Bacteria ; drug effects ; Caryophyllaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology

OBJECTIVETo study the chemical constituents of Psammosilene tunicoides.

METHODThe two chemical constituents were separated by various chromatographic methods, and their structures were identified on the basis of spectroscopic analysis.

RESULTTwo beta-carboline alkaloids were separated from normal butanol fraction of P. tunicoides, and identified as 1-acetyl-3-methoxycarbonyl-beta-carboline (1) and 1-acetyl-3-methoxycarbonyl-4-hydroxyl-beta-carboline (2).

CONCLUSIONCompound 1 is separated from this plant as natural products for the first time, and compound 2 was a new compound.

Alkaloids ; analysis ; Carbolines ; analysis ; Caryophyllaceae ; chemistry

The total triterpene saponins of Psammosilene tunicoides have significant pharmacologic activity. Psammosilene tunicoides squalene synthase (PSS) is a gateway enzyme to regulate the biosynthesis of total triterpene saponins extracted from the root of Psammosilene tunicoides which is an endangered species. In this paper, cDNA encoding of PSS was cloned by the degenerate primer PCR and rapid-amplification of cDNA ends (RACE). The full-length of cDNA of PSS is 1663 bp, with an open reading frame (ORF) of 1 245 bp, encoding 414 amino acid polypeptide (calculated molecular mass, 47.69 kDa), 5'UTR (untranslated region) and 3'UTR are 260 bp and 158 bp, respectively. The deduced amino acid sequence of PSS has higher homology with the known squalene synthases of several species such as Panax notoginseng (83%), Panax ginseng (82%) and Glycyrrhiza glabra (82%) than that with Schizosacharomyces pombe (35%), Candida albicans (39%) and Homo sapiens (47%). The characterization of PSS was done by a series of methods, such as prokaryotic expression, the activity of enzyme in vitro, capillary gas chromatography (GC) and capillary gas chromatography mass spectrometry (GC-MS). The results showed that the cell-free extract of E. coli transformed with the recombinant plasmid can effectively convert farnesyl diphosphate into squalene in vitro. GenBank accession number is EF585250. Our research provided important base for the study of Psammosilene tunicoides secondary metabolism and metabolic engineering.
Caryophyllaceae ; enzymology ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Endangered Species ; Escherichia coli ; genetics ; metabolism ; Farnesyl-Diphosphate Farnesyltransferase ; genetics ; metabolism ; Gas Chromatography-Mass Spectrometry ; Open Reading Frames ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; metabolism ; Sequence Homology, Amino Acid ; Transformation, Genetic