This study examined the chemical composition of A. blasiliensis and the chemical structural properties of an immuno-stimulating polysaccharide. The amino acids, free sugars, and organic acids by HPLC and fatty acids by GC were analyzed. The immuno-stimulating substance from A. blasiliensis was extracted with hot water and purified by ethanol precipitation. It underwent ion exchange chromatography on DEAE-cellulose and gel filtration on Toyopearl HW 65F. Through GP-HPLC, the substance was found to be homogeneous. Its chemical structure was determined by 13C-NMR. Fatty acids, organic acids, and sugar alcohol composition consisted exclusively of linoleic acid, fumaric acid and mannitol, respectively. The amino acids were mainly glutamic acid, glycine, and arginine. By 13C-NMR analysis, the immuno-stimulating substance was identified as beta-(1-->3) (1-->6)-glucan, composed of a backbone with (1-->3)-linked D-glucopyranosyl residues branching a (1-->6)-linked D-glucopyranosyl residue. The beta-glucan from A. blasiliensis showed pronounced immuno-stimulating activity on the antibody-production ability of B-lymphocytes by the hemolytic suspension assay. In these results, A. blasiliensis was estimated to have potent pharmacological properties and potential nutritional values.
Agaricus ; Amino Acids ; Arginine ; B-Lymphocytes ; Carbohydrates ; Chromatography, Gel ; Chromatography, High Pressure Liquid ; Chromatography, Ion Exchange ; DEAE-Cellulose ; Ethanol ; Fatty Acids ; Fumarates ; Glutamic Acid ; Glycine ; Linoleic Acid ; Mannitol ; Nutritive Value ; Water

Two new azaphilone derivatives containing 1,3-dioxolane moiety, penidioxolanes A (1) and B (2), were isolated from marine-derived fungus Penicillium sp. KCB12C078, together with four known compounds (3-6) by chemical investigation. Compounds 1 - 6 were isolated by combination of silica gel, ODS column chromatography and preparative HPLC. Their structures were determined by analysis of spectroscopic data including 1D-, 2D-NMR, and MS techniques. The isolates were evaluated against cancer cell growth inhibition effects and antimicrobial activity.
Chromatography ; Chromatography, High Pressure Liquid ; Fungi ; Penicillium* ; Silica Gel

Exopolysaccharide La0.1-1 was extracted from the broth of a marine bacterium Lentibacter algarum ZXM100T isolated from the seawater in the coastal region of Qingdao and purified by Q Sepharose Fast Flow ion-exchange chromatography and Superdex 75 gel-permeation chromatography. Its physiochemical properties and primary structural characters were investigated by chemical analysis together with high performance liquid chromatography (HPLC), high performance gel permeation chromatography (HPGPC) and gas chromatography and mass spectrometry (GC-MS). The results show that the total sugar content of the exoploysaccharide La0.1-1 was about 66% with an average molecular weight at 12.0 kDa. La0.1-1 is mainly composed of Gal, Man, GlcN at the ratio of 1.35:1.1:1.0. Results of GC-MS and NMR demonstrate that the exopolysaccharide La0.1-1 mainly exists with the beta configuration. The primary linkage styles are --> 2)-Manp(1 --> and --> 3)-Galp(1 --> with a small amount of --> 4)-Galp(--> 1 and --> 4)-Manp(1 --> linkages. The linkage mode of GlcN is --> 4)GlcN(1 --> and terminal linkage. The exopolysaccharide has mainly a linear sructure with a few branches linked to 0-6 of --> 2)-Manp(1 --> and 0-4 or 0-6 of --> 3)-Galp(1 -->. 1D-NMR data also revealed that La0.1-1 is substituted by certain acetyl; the acetyl is mainly linked to N-2 of GlcN. The exopolysaccharides of the bacterium of Lentibacter genus is reported for the first time, and an exopolysaccharide with novel structure was obtained, which enriched marine polysaccharide resources.
Chromatography, Gel ; Chromatography, High Pressure Liquid ; Chromatography, Ion Exchange ; Gas Chromatography-Mass Spectrometry ; Magnetic Resonance Spectroscopy ; Molecular Weight ; Polysaccharides ; chemistry ; isolation & purification ; Rhodobacteraceae ; chemistry ; Seawater ; microbiology

OBJECTIVETo provide application basis of the Forsythia suspensa by studying the difference of HPLC-FP of F. suspensa fructification (medicinal materials).

METHODComparative work was done on F. suspensa produced in different areas, on different parts of Forsythia suspensa, and on the pseudo preducts with methods of HPLC-FP.

RESULTDifferent FP characteristics were shown respectively by different samples, which were from different producing areas, from different parts, and the pseudo products including the fructification of Syringa reticulata var. and F. viridissimac.

CONCLUSIONThe FP can be used to distinguish the F. suspensa coming from different producing areas and different sources.

Chromatography, High Pressure Liquid ; Chromatography, Paper ; methods ; Drug Contamination ; Drugs, Chinese Herbal ; chemistry ; Forsythia ; chemistry ; classification ; Fruit ; chemistry ; Peptide Mapping ; Plants, Medicinal ; chemistry ; Species Specificity

Globally, peptide-based anticancer therapies have drawn much attention. Marine organisms are a reservoir of anticancer peptides that await discovery. In this study, we aimed to identify cytotoxic oligopeptides from Sarcophyton glaucum. Peptides were purified from among the S. glaucum hydrolysates produced by alcalase, chymotrypsin, papain, and trypsin, guided by a methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay on the human cervical cancer (HeLa) cell line for cytotoxicity evaluation. Purification techniques adopted were membrane ultrafiltration, gel filtration chromatography, solid phase extraction (SPE), and reversed-phase high-performance liquid chromatography (RP-HPLC). Purified peptides were identified by de novo peptide sequencing. From papain hydrolysate, three peptide sequences were identified: AGAPGG, AERQ, and RDTQ (428.45, 502.53, and 518.53 Da, respectively). Peptides synthesized from these sequences exhibited cytotoxicity on HeLa cells with median effect concentration (EC₅₀) values of 8.6, 4.9, and 5.6 mmol/L, respectively, up to 5.8-fold stronger than the anticancer drug 5-fluorouracil. When tested at their respective EC₅₀, AGAPGG, AERQ, and RDTQ showed only 16%, 25%, and 11% cytotoxicity to non-cancerous Hek293 cells, respectively. In conclusion, AERQ, AGAPGG, and RDTQ are promising candidates for future development as peptide-based anticancer drugs.
Amino Acid Sequence ; Animals ; Anthozoa/chemistry* ; Antineoplastic Agents/pharmacology* ; Chromatography, Gel ; Chromatography, High Pressure Liquid ; Chromatography, Reverse-Phase ; Cytotoxins/pharmacology* ; Drug Discovery ; HEK293 Cells ; HeLa Cells ; Humans ; Hydrolysis ; Marine Toxins/pharmacology* ; Oligopeptides/pharmacology* ; Solid Phase Extraction ; Tandem Mass Spectrometry

OBJECTIVES: This study was conducted in order to understand the cellular events associated with silica-induced pathogenesis of the rat lung. METHODS: Silicosis was induced by an intratracheal instillation of 50 mg of silica (SiO2, 0.15 - 10 micrometer) suspended in 500 microliter of a sterile saline solution in Sprague-Dawley rats weighing 200g. Silicotic nodules were excised from the rat lungs 4 weeks after silica instillation, then boiled for 4 days at 110 degrees in solution containing 2% SDS, 10 M urea and 40 mM DTT. The insoluble cellular encapsulates were electrophoresed on 4-12 % gradient SDS-PAGE, and the amino acid composition was analyzed. Affinity chromatographies of the homogenate supernatants of the control lung, silicotic nodule, and normal rat plasma were performed using rabbit IgG, anti-rat, cross-linked protein from the silicotic nodule. The amounts of N epsilon-(gamma-glutamyl) lysine cross-linked in the control lungs and silicotic nodules were determined using HPLC analysis. RESULTS: The remaining cross-linked protein was insoluble in the 10 M urea and 40 mM sulfhydryl reagents even under prolonged boiling conditions. The encapsulate revealed the retention of silica particles within the protein whose amino acid composition showed a high percentage of alanine, leucine and glycine. A 46 KDa protein was identified as a cross-linked protein in the silicotic nodule by affinity chromatography. The level of N epsilon-(gamma-glutamyl) lysine dipeptide in the nodule digest was prominently increased compared with that in the control lung. CONCLUSIONS: Transglutaminase (TGase)-catalyzed cross-linking appears to be involved in the silicotic nodule formation, and the 46 KDa protein may be cross-linked to itself and other extracellular matrix proteins during fibrosis and the formation of eventually insoluble nodule.
Alanine ; Animals ; Chromatography ; Chromatography, Affinity ; Chromatography, High Pressure Liquid ; Electrophoresis, Polyacrylamide Gel ; Extracellular Matrix Proteins ; Fibrosis ; Glycine ; Immunoglobulin G ; Leucine ; Lung* ; Lysine ; Plasma ; Rats* ; Rats, Sprague-Dawley ; Silicon Dioxide ; Silicosis ; Sodium Chloride ; Sulfhydryl Reagents ; Urea

Human cytomegalovirus (HCMV)-specific monoclonal antibody, SCMVM 34, recognizes early antigen confined to the nucleus of HCMV-infected cells. This study was performed to identify the antigen reactive to SCMVM 34 with purification and amino acid sequencing. The nuclear and cytoskeletal fraction of HCMV-infected cells was subjected to 0.4 M NaCl extraction, DEAE-Sephacel ion exchange chromatography, DNA-cellulose chromatography and SDS-PAGE. The molecular weight of the reactive proteins was 52 kD, 40 kD and 34 kD. The modified or blocked amino termini of 52 kD and 40 kD showed resistance to Edman degradation. The internal peptide fragments were isolated by tryptic digeytion and reverse-phase HPLC. The internal amino acid sequence analysis of the peptides from HPLC profile revealed that the antigens recognized by SCMVM 34 was ppUIA4.
Chromatography ; Chromatography, High Pressure Liquid ; Chromatography, Ion Exchange ; Cytomegalovirus* ; Electrophoresis, Polyacrylamide Gel ; Humans* ; Molecular Weight ; Peptide Fragments ; Peptides ; Sequence Analysis, Protein

The fingerprint technology could reflect the internal chemical characteristics of Chinese herbal medicine or preparation, which has the characteristics of "wholeness" and "fuzziness". It is suitable for evaluating the quality of intermediate and finished products in the production process of traditional Chinese medicine formula granules. In this paper, the applications of high performance liquid chromatography (HPLC), thin layer chromatography (TLC), gas chromatography (GC) and infrared spectrum (IR) fingerprint technology in the quality control of traditional Chinese medicine formula granules were reviewed, and their advantages and disadvantages were analyzed. The aim of this article is to enhance the combined application of various fingerprint technologies in traditional Chinese medicine formula granules. It could provide technical reference for realizing the stability of production process and improving the overall quality of formula granules.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; standards ; Medicine, Chinese Traditional ; Quality Control

Eleven compounds were isolated from the 95% ethanol extract of the stems of Dendrobium officinale after water extraction by various modern chromatographic techniques, such as silica gel column chromatography(CC), octadecyl-silica(ODS) CC, Sephadex LH-20 CC, preparative thin layer chromatography(PTLC) and preparative high performance liquid chromatography(PHPLC). According to spectroscopic analyses(MS, 1D-NMR, 2D-NMR) combined with optical rotation data and calculated electronic circular dichroism(ECD), their structures were identified as dendrocandin Y(1), 4,4'-dihydroxybibenzyl(2), 3-hydroxy-4',5-dimethoxybibenzyl(3), 3,3'-dihydroxy-5-methoxybibenzyl(4), 3-hydroxy-3',4',5-trimethoxybibenzyl(5), crepidatin(6), alternariol(7), 4-hydroxy-3-methoxypropiophenone(8), 3-hydroxy-4,5-dimethoxypropiophenone(9), auriculatum A(10) and hyperalcohol(11). Among them, compound 1 was a new bibenzyl derivative; compounds 2 and 7-11 have not been previously reported from Dendrobium plants; compound 6 was reported from D.officinale for the first time. Compounds 3-6 exhibited potent antioxidant activity with IC_(50) values of 3.11-9.05 μmol·L~(-1) in ABTS radical scavenging assay. Compound 4 showed significant inhibitory effect on α-glucosidase, with IC_(50) value of 17.42 μmol·L~(-1), indicating that it boasted hypoglycemic activity.
Dendrobium ; Biological Assay ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Bibenzyls

Based on the approaches of TLC identification, HPLC for assaying managiferin, and of the determination of water, total ash and acid-insoluble ash in 12 samples, collected from southwest of China, the quality standard of Gentiana rhodantha has been established. The results show the reference materia medica and mangiferin can be both well used as reference substances for TLC identification; the mass fractions of mangiferin is 0.7%-4.4% (average 2.8%), water 6.1%-8.6% (average 7.2%), total ash 3.7%-10.8% (average 6.6%) and acid-insoluble ash 0.2%-2.7% (average 1.3%). The recommended standards of quantitative indexes are that the mass fractions of mangiferin is not less than 2.0%, and the water, total ash and acid-insoluble ash are not more than 9.0%, 8.0% and 3.0% respectively.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; analysis ; Gentiana ; chemistry ; Reproducibility of Results