Determination of fat-soluble chlorinated compounds in fish

Summary The level of chlorinated non-polar compounds in samples of sprat (Clupea sprattus) has been determined. The total amounts of chlorine, bromine and iodine in the lipid phase were determined by neutron activation analysis prior to and after treatment of the lipid phase with concentrated sulfuric acid. In sprat samples from a contaminated area the following chlorinated hydrocarbons have been identified and quantified by means of gas chromatography and mass spectrometry: pentachlorobenzene, hexachlorobenzene, heptachlorostyrene and octachlorostyrene. In addition, the following compounds have been detected by gas chromatography using standards: dichlorobenzene, trichlorobenzene, tetrachlorobenzene and polychlorinated biphenyls. A gradual reduction of the level of individual components and total level of chemically persistent chlorinated hydrocarbons in the samples is observed with increasing distance from the source of contamination (Frierfjorden). The same tendency is not seen for the PCB. It has been found that highly chlorinated compounds are distinctly enriched in the sprat oil relative to compounds with lower contents of chlorine.

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Bestimmung von fettlöslichen chlorierten Verbindungen in Fisch

Zusammenfassung Chlorierte unpolare Verbindungen wurden in Fischproben (Sprotten, Clupea sprattus) bestimmt. Der Gesamtgehalt an Chlor, Brom und Jod in der Fettphase wurde vor und nach deren Behandlung mit konz. Schwefelsäure mit Hilfe der Neutronenaktivierungsanalyse erfaßt. In Proben aus einem kontaminierten Gebiet wurden folgende Verbindungen durch Gas-Chromatographie/ Massenspektrometrie identifiziert und quantitativ bestimmt: Pentachlorbenzol, Hexachlorbenzol, Heptachlorstyrol und Octachlorstyrol. Zusätzlich wurden die folgenden Verbindungen durch GC mit Hilfe von Standards erfaßt: Dichlorbenzol, Trichlorbenzol, Tetrachlorbenzol, polychlorierte Biphenyle. Eine allmähliche Abnahme des Gehaltes an einzelnen Verbindungen sowie des Gesamtgehaltes an beständigen chlorierten Kohlenwasserstoffen wurde mit zunehmender Entfernung vom Ursprung der Verschmutzung (Frierfjorden) beobachtet. Dies konnte jedoch im Falle von PCBs nicht festgestellt werden. Hochchlorierte Verbindungen werden im Sprottenöl im Gegensatz zu solchen mit niederen Chlorgehalten deutlich angereichert.

Presented at the 6th Annual Symposium on Recent Advances in the Analytical Chemistry of Pollutants, April 21–23, 1976; Vienna, Austria.     

Flavonoids of Roots of Glycyrrhiza uralensis Growing in Siberia

The composition of phenolic components of roots and rhizomes ofGlycyrrhiza uralensisFisch. from Siberia is studied. A total of 15 components belonging to 8 structural types including 4' -methylglucoliquiritigenin, which was previously unknown in licorices, is found. The components in samples collected from various regions of Siberia are practically identical.     

Baseline studies of the global pollution

Summary Fish and fish oils, which can be assigned to defined food webs, habitats and regions in the North Atlantic (Gulf Stream, Sargasso Sea, continental shelf of Iceland, Ireland, Norway, North Sea and Portugal) have been analyzed for organochloro compounds like hexachlorocyclohexane isomers (HCH), chloroterpene (toxaphen, polychlorocamphene), cyclodien pesticides (chlordane group, dieldrin), chlorobenzenes, chlorobiphenyls (PCB) and the DDT group. Phytoplancton feeding fish menhaden (Brevoortia tyrannus) were used to characterize the surface layer of the Atlantic west of the United States, while ground feeding predatory cod (Gadus morhua) and sea pike (Merluccius merluccius) should picture mainly the pollution near the continental shelf at 200–600 m depth.All groups of chemicals listed above could be found in the fish samples and in most cases the single components of mixtures (PCB, chlordane) could be identified by high resolution glass capillary gas chromatography with electron capture detection. Intensity and details of the pollution pattern vary strongly for the water regions investigated. The global water cycle and details of pollution input and history of water bodies can be used for an explanation.

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Untersuchungen zur globalen Belastung der UmweltIV. Belastungsmuster der Organochlorverbindungen im Nordatlantik, angereichert in Fischen

Zusammenfassung In Fischen, die einer bestimmten Nahrungskette und Region zuzuordnen sind (Bermuda, Kontinentalschelf westlich von Irland und Portugal) und in Dorschleberölen, die aus definierten Regionen des Nordatlantiks (Kontinentschelf von Island, Irland, Norwegen, Nordsee und Portugal) stammen, wurde die Belastung mit Hexachlorcyclohexanen (HCH), Chlorterpenen (Toxaphen), Cyclodien-Biociden (Chlordan-Gruppe, Dieldrin), Chlorbenzolen, Chlorbiphenylen (PCB) und der DDT-Gruppe bestimmt. Im Oberflächenwasser lebende Phytoplanktonfiltrierer [Menhaden (Brevoortia tyrannus)] wie auf dem Schelfsockel lebende bathypelagische Räuber [Dorsch, Kabeljau (Gadus morhua) und Seehecht (Merluccius merluccius)] sind untersucht worden. Damit ist der Eintrag sowohl nahezu unmittelbar aus dem Wasser (Phytoplanktonfiltrierer) wie über eine längere mittelbare und unmittelbare Nahrungskette (bathypelagische Räuber), erfaßt worden. Alle aufgeführten Substanzklassen konnten z. T. mit weitgehender Identifizierung der Einzelkomponenten durch hochauflösende Glas-Capillar-Gas-Chromatographie und Elektroneneinfang-Detektion nachgewiesen werden. Es zeigen sich deutliche regionale Unterschiede in der Stärke der Belastung und in dem Belastungsmuster. Die globale Wasserführung und der damit verbundene unterschiedliche Eintrag wie die unterschiedliche Historie der Wassermassen können als Erklärung herangezogen werden.

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List of Abbreviations PCBZ Polychlorobenzene - TrCBZ Trichlorobenzene - PeCBZ Pentachlorobenzene - HCH Hexachlorocyclohexane - PCB Polychlorobiphenyl - 1-209 Polychlorobiphenyl of defined structure [4] - B Polychlorobiphenyl of undefined structure - DDT Dichlorodiphenyl-trichloroethane - 2,4-DDT 1,1,1-Trichloro-2-(2-chlorophenyl)-2-(4-chlorophenyl)-ethane - 2,4-DDD 1,1-Dichloro-2-(2-chlorphenyl)-2-(4-chlorophenyl)-ethane - 2,4-DMD 1-Chloro-2-(2-chlorophenyl)-2-(4-chlorophenyl)-ethane - 2,4-DDE 1,1-Dichloro-2-(2-chlorophenyl)-2-(4-chlorophenyl)-ethene - 2,4-DDMU 1-Chloro-2-(2-chlorophenyl)-2-(4-chlorophenyl)-ethene - 2,4-DBP 2,4-Dichloro-benzophenone - 4,4-DDT 1,1,1-Trichloro-2,2-bis(4-chlorophenyl)-ethane - 4,4-DDD 1,1-Dichloro-2,2-bis(4-chlorophenyl)-ethane - 4,4-DMD 1-Chloro-2,2-bis(4-chlorophenyl)-ethane - 4,4-DDE 1,1-Dichloro-2,2-bis(4-chlorphenyl)ethene - 4,4-DDMU 1-Chloro-2,2-bis(4-chlorophenyl)-ethene - 4,4-DBP 4,4-Dichloro-benzophenone - CY Dieldrin Endrin Cyclodien-biocides - Hepo Heptachloroepoxid - CY 40 cis-Chlordane - CY 41 trans-Chlordane - CY 50 cis-Nonachlor - CY 51 trans-Nonachlor - PCT(T) Polychloroterpene (Toxaphene, Polychlorocamphen, Strobane) - U Major unknowns/not identified compounds     

甘草真伪品的FTIR光谱法鉴别研究

采用傅里叶变换红外(FTIR)光谱法并结合二维相关(Two-dimensional correlation spectroscopy)分析技术分别对药用植物甘草(药材对照品)及其伪品刺果甘草进行了无损快速鉴别研究。 结果表明：虽然甘草和伪品刺果甘草都是来源于同一科属,但两者所含化学组分的含量不同,其红外光谱图既有一定的差异,又有一定的相似。 而在红外二阶导数谱图上差异较明显,在二维红外谱图的差别不但较明显而且很直观。 凭借这些差异,可方便地进行真伪鉴别,同时还进一步表明了这两种甘草药材中化学组分之间的差异。 该法无损,快速,准确,样品量少,为客观评价中药材的来源,真伪,优劣等提供了一种新的方法和手段。     

Simultaneous separation of triterpenoid saponins and flavonoid glycosides from the roots of Glycyrrhiza uralensis Fisch by pH‐zone‐refining counter‐current chromatography

Glycosides including triterpenoid saponins and flavonoid glycosides are the main constituents of Glycyrrhiza uralensis Fisch (licorice) and exhibit prominent pharmacological activities. However, conventional methods for the separation of glycosides always cause irreversible adsorption and unavoidable loss of sample due to their high hydrophilicities. The present paper describes a convenient method for the simultaneous separation of triterpenoid saponins and flavonoid glycosides from licorice by pH‐zone‐refining counter‐current chromatography. Ethyl acetate/n‐butanol/water (2:3:5, v/v) with 10 mM TFA in the upper organic stationary phase and 10 mM ammonia in the lower aqueous mobile phase was used as the biphasic solvent system. Three triterpenoid saponins and two flavonoid glycosides including licorice‐saponin A3 (63.3 mg), glycyrrhizic acid (342.2 mg), 3‐O‐[β‐d ‐glucuronopyranosyl‐(1 → 2)‐β‐d ‐galactopyranosyl]glycyrrhetic acid (56.0 mg), liquiritin apioside (232.6 mg), and liquiritin (386.5 mg) were successfully obtained from licorice ethanol extract (2 g) in one step. This method subtly takes advantage of the common acidic properties of triterpenoid saponins and flavonoid glycosides, and obviously is much more efficient and convenient than the previous methods. It is also the first time that the separation of acidic triterpenoid saponins by using pH‐zone‐refining counter‐current chromatography has been reported.     

Synthesis of amino-silane modified magnetic silica adsorbents and application for adsorption of flavonoids from Glycyrrhiza uralensis Fisch

Magnetic separation technology was applied in the separation of flavonoids from the licorice root in this work. Licorice flavonoids (LF) displayed a remarkable array of biological and pharmacological activities. The magnetic adsorbents with functional -NH2 groups were synthesized by immobilization of amino-silane on the surface of the magnetic silica supports, which were prepared by co-precipitation method. The adsorption and desorption characteristics of the magnetic adsorbents for the separation of LF have been evaluated. The purity of an enriched extract with this method was 16.7% while the crude extract only had about 6.8% purity. Therefore, it can be concluded that these kinds of magnetic adsorbents have selectivity to the flavonoids to some extent. The affinity selectivity of the adsorbents is based on the formation of hydrogen bonding between the -NH2 on the magnetic adsorbents and -OH,-CO on the flavonoids.     

Tracking antiangiogenic components from Glycyrrhiza uralensis Fisch. based on zebrafish assays using high-speed countercurrent chromatography

Natural products are some of the most important sources of lead compounds for drug discovery. The advanced isolation technique of lead compounds of natural origin using therapeutically relevant bioassays is capable of enhancing work efficiency from complex multiconstituent extracts. In the present study, a bioassay-guided isolation strategy combined with bioactivity screening was used to identify novel angiogenesis inhibitors from licorice (Glycyrrhiza uralensis Fisch.) based on the zebrafish model and rapid preparative separation by high-speed countercurrent chromatography. Zebrafish embryos at 24 h postfertilization were chosen as the angiogenesis inhibition model for bioactivity screening. A solvent system (n-hexane-ethyl acetate-methanol-water) with different ratios was optimized and applied in the high-speed countercurrent chromatography separation of two fractions, Fr5 and Fr6, from the ethyl acetate extract of licorice. Blood circulation and vascular outgrowth in intersegmental vessels were found to be simultaneously inhibited by isoliquiritigenin and isolicoflavonol in a dose-dependent manner. Thus, these two compounds were identified and considered as active inhibitors against angiogenesis. These experimental results indicate that zebrafish bioassays combined with high-speed countercurrent chromatography may provide an alternative pathway for the rapid isolation of bioactive natural products.