Isolation of lipids from photosystem I complex and its characterization with high performance liquid chromatography/electrospray ionization mass spectrometry.

A method for simultaneous analysis of lipids extracted from photosystem I complex was developed with high performance liquid chromatography/electrospray ionization mass spectrometry. The photosystem I complex was firstly solubilized and separated using deoxycholate polyacrylamide gel electrophoresis method after ultrasonic treatment of the sample (leaves of pea, Pisum sativum L.). The Photosystem I complexes were electrophoretically eluted from the deoxycholate polyacrylamide gel electrophoresis bands containing them, and the electron transport activity of the eluent measured as confirmation. Lipids, which were isolated from the complex having photosystem I activity, were separated and characterized with high performance liquid chromatography/electrospray ionization mass spectrometry. Five lipids, monogalactosyldiacylglycerol, digalactosyldiacylglycerol, phosphatidylglycerol, sulphoquinovosyldiacylglycerol and phosphaditylcholine were found combining with photosystem I complex. Different species of these lipids were found in the ESI mass spectra and the compositions of the acyl groups in them were determined.

Enantioseparation of 2-O-beta-D-glucopyranosyl-2H-1,4-benzoxazin-3(4H)-one and its 7-chloro-derivative by capillary zone electrophoresis using native and substituted beta-cyclodextrins as chiral additives.

Efficient, rapid and inexpensive methods were established for the chiral separation of two glucopyranosyl compounds from plant extracts, by capillary zone electrophoresis (CZE). Baseline separation was achieved for both compounds. Several native cyclodextrins and their derivatives were tried as chiral selectors. CM-beta-CD and HP-beta-CD (with addition of acetonitrile in the buffer) gave rise to optimal chiral separation for the two compounds, respectively, each within a few minutes. The effects of several parameters on the chiral separation were studied.

Analysis and determination of biological activity of short-chain peptides from porcine brain hydrolysate.

Gel permeation chromatography fractions of short-chain peptides from a hydrolysate product, which in turn was from the purified porcine brain through enzyme hydrolysis, were tested for their biological activities. The results showed that the fractions A4 and A5 had significant biological activities. The two fractions were analyzed with analytical techniques such as high-performance liquid chromatography (HPLC), capillary zone electrophoresis (CZE), isoelectric focusing (IEF), discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Preliminary results showed that the main components of these two fractions were short-chain acidic peptides with a relative molecular mass (M(r)) of less than 2400.

Comparative study of the composition of TCM Radix notoginseng extract samples before and after acetylation with UV and ELSD detection.

OBJECTIVE: To study the derivation, acetylation specifically, as well as the application of evaporative light-scattering detector (ELSD) detection of Panax notoginseng extract for a better understanding of its components, as well as to study the authentication of this Traditional Chinese Medicine (TCM). METHODS: Acetonitrile-water gradient elution of the samples was used for the analysis and the ultraviolet (UV) detection used to observe the difference between the extract samples before and after acetylation. Mobile phases containing 30% and 85% acetonitrile, respectively, were used to observe the differences between chromatograms of the samples obtained using UV and ELSD detection. RESULTS: By acetylating the extract before analysis, differentiation of the early-eluting components was observed, some of the derivatives were retained extremely strongly. Different eluting profiles were obtained from the extract samples using UV and ELSD. Using the latter technique, different patterns of change in the retention of peaks could be observed, uncovering more information relating to the composition of the extract. CONCLUSION: The decrease of polarities of a part of the hydrophilic components as a result of acetylation of the extract and the differentiation of these early-eluting, difficult-to-separate compounds in the chromatograms should be helpful for the characterization and authentication of the TCM. ELSD can be used to detect the carbohydrates, which are known to have pharmacological effects, and sensitize the detection of glycosides. This is also helpful for the above-mentioned aspects.

Exact electric field strength over the packed capillary electrochromatography column bed, theoretical considerations and practical determination.

On the basis of the experiments, it was proved that Ohm's law holds true in a capillary electrochromatography (CEC) column. By using the additivity of the potential drop over the packed and unpacked sections, the exact values of the electric field strength over the packed CEC column bed were determined experimentally. The ratio of the resistance of packed CEC column sections to that of an open capillary of the same length (Rp/Ro,Lp) were calculated. Some theoretical calculations were made to show how some structural parameters of particle lattice affect the resulting Rp/Ro,Lp value of the column bed formed accordingly. It is suggested that to correctly elucidate the EOF phenomenon in a CEC column, the potential drop over the packed and unpacked column sections, Ep and Eo, should be accounted for respectively, rather than using the average field strength over the total column length.

Chromatographic behavior of packing materials for capillary electrochromatography with a coating of different immobilized polysiloxanes.

Octadecyl silyl silica (ODS) phase coated with immobilized polysiloxanes (OV1701, SE-54, SE-30) were synthesized, their characteristics as capillary electrochromatography (CEC) column packing materials were studied. It was found that, although the polysiloxane coatings were different in polarity, the resulting packing materials showed the highest efficiencies when the respective coating ratios (polysiloxane:ODS, w/w) were all 20-30%. As expected, packing materials coated with different polysiloxanes resulted in different selectivity on solute pairs. Separations on these stationary phases were studied with different factors such as pH values and acetonitrile contents of the mobile phases. It was found that all these kind of stationary phases could resist basic mobile phase with a pH value as high as 11.6. Tests were made to analyze polar, basic drugs with CEC using the stationary phases.

Electrochromatographic properties of synthetic poly(styrene-divinylbenzene) encapsulated packing material and sulphonic cation-exchange phase based on the former.

OBJECTIVE: To develop new kinds of stationary phases suitable for applications in capillary electrochromatography (CEC), as well as in LC, which can be tailored to realize selective separations of solutes difficult to separate with conventional stationary phases. METHODS: Purchased spherical silica particles (5 microm) were refluxed in anhydrous toluene with vinyl-triethoxysilane for 18 h, to modify the surface with vinylsilyl groups. The silylated silica particles were subsequently stirred in an n-henanol-acetonitril mixture (1:4, volume fraction) for 18 h. Styrene and divinylbenzene in the mixture co-polymerized to form a crosslinked encapsulation layer on the silica surface, and bonded on the surface vinyl groups at the same time, both reactions were initiated by azo-iso-butyronitrile. The poly(Styrene-divinylbenzene) encapsulated silica(PS-DES) produced can be used as a non-conventional stationary phase for CEC itself. By sulphonation of the PS-DES phase with chlorosulphonic acid, strong cation-exchange stationary phase of sulphonic type was obtained. RESULTS: The manufactured PS-DES as well as the sulphonated phases were respectively packed into capillaries,and the columns thus prepared were tested for their chromatographic characteristics. It was found that the PS-DES phase showed reversed-phase characteristics. Due to the phenyl groups in the encapsulated polymer, it introduced pi-pi electronic interaction between the solutes molecules and stationary phase during the chromatographic separation process, therefore it showed unique selectivity on separating aromatic compounds, also polar- as well as some alkaline drugs was analyzed on the column packed with the phase. It was demonstrated that the sulphonated phase could be used to prepare columns for the separation of alkaline drugs, symmetric peaks were obtained for them and base-line separation was realized. CONCLUSION: It is possible that these stationary phases prepared can be used for solving the analytical problems in which non-conventional selectivity are needed ( the analyses of alkaline drugs are examples) In some cases, the analysts can realize the anticipated separation results based on the different separation mechanism from that of the conventional stationary phases.