Improved Assay For 6-Keto Prostaglandin F1α In Biological Fluids By Reversed-Phase High-Performance Liquid Chromatography In Conjunction With Radioimmunoassay

1981 ◽  
Author(s):  
P B B Jones ◽  
J D Wark ◽  
R G G Russell
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Biological Fluids ◽  
Ultra Violet ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Breakdown Product ◽  
Radial Compression ◽  
Good Resolution

Prostacyclin production has been proposed as a fundamental mechanism by which intact vascular endothelium maintains a non-thrombogenic surface. Interest in a reliable assay for its stable breakdown product 6-keto prostaglandin F1α (6-KPGF1α) is, therefore, considerable. Previous studies have emphasised the need to reduce sample complexity prior to the quantification of PG’s by radioimmunoassay (RIA). This has been achieved by extraction of PG’s into organic solvents or onto resin columns followed by separation of PG metabolites by reversed-phase high-performance liquid chromatography (HPLC), but retention times are long if adequate resolution is to be obtained. By using an 8-min gradient elution programme with acetonitrile/water/acetic acid and a 6 ml/min flow rate through Waters reversed-phase (Radial Pak A) column under radial compression good separation of 6-KPGF1α from its closely-related metabolites 13,14-dihydro 6-KPGF1α 6, 15-diketo PGF1α as well as from other PG species was achieved as detected by ultra-violet absorbance spectrometry. In applying the method to the measurement of 6-KPGF1α in urine, PG’s were extracted onto a resin column, eluted in methanol and the extract subjected to HPLC as described. Good resolution of immunoassayable PG metabolites and known cross-reacting species was achieved. This method provides a significant improvement on existing PG assays by reducing sample-processing time dramatically, and is suitable for routine and repetitive clinical use.

High Performance Liquid Chromatography Coupled with Radioactivity Detection: A Powerful Tool for Determining Drug Metabolite Profiles in Biological Fluids

1986 ◽  
Vol 41 (1-2) ◽  
pp. 115-125 ◽  
Author(s):  
K.-O. Vollmer ◽  
W. Klemisch ◽  
A. von Hodenberg
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Biological Fluids ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Isolation And Purification ◽  
Carbon 14 ◽  
Tracer Techniques ◽  
Metabolite Profiles

Abstract High performance liquid chromatography coupled with continuous radioactivity detection rep­resents an advancement in drug metabolism research. Using radioactive substances labelled in biologically stable positions, all metabolites can be specifically detected by radioactivity measure­ment. Thus no clean-up of biological fluids is required prior to HPLC. This can prevent artefact formation from unstable metabolites, reduces recovery problems and facilitates quantitation. Separation of highly polar and unpolar metabolites is possible in a single chromatographic run using gradient elution and reversed phase materials. This technique is also well-suited for prepara­tive isolation and purification of metabolites for subsequent structure elucidation. Various metabolite profiles of drugs labelled with carbon-14 or tritium are shown. Metabolites of the following drugs are presented: norfenefrine, etozolin, thymoxamine, naloxone, and levobunolol. We review the general methodology and report our experience with this technique. In principle, this technique may be useful for all biological systems in which tracer techniques are applied.

Analysis of Pandanus Extracts by Reversed Phase High Performance Liquid Chromatography

2016 ◽  
Vol 13 (2) ◽  
pp. 1
Author(s):  
Hannis Fadzillah Mohsin ◽  
Ratni Suriyani Jalal ◽  
Ibtisam Abdul Wahab ◽  
Abdul Rasyid Zulkifli ◽  
Muhammad Faxrurrazy Sadiran
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Ultra Violet ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Automated System ◽  
Elution Time ◽  
Research Areas ◽  
Alcoholic Extraction

The Pandanus species (Pandanaceae family) was investigated, in order to set up a library of their chromatographic profiles. From a literature review, the most common Pandanus plant that was greatly examined was P. amaryllifolius (the fragrant screw pine), followed by P. sanderi. Here, the chromatographic data from an extract of a relatively unfamiliar Pandanus is also presented. The leaves of P. monotheca were extracted by using methanol. Later, the extract was filtered, prior to a Reversed-Phase High Performance Liquid Chromatography (RP-HPLC) experiments. The automated system was set to run by gradient elution (acetonitrile:water). The flow rate was maintained at 1 ml/min, otherwise stated in a number of attempts. The ultra-violet absorbance of the Pandanus’ constituents was measured via Multiple Wavelength Detector. From the chromatogram, the major peaks for Pandanus’ compounds were eluted (λ = 220, 260 and 360 nm) and the retention times were recorded. The lower absorbance readings of other minor peaks could be due to various minor secondary metabolites, such as the furolignan. The different elution time depends on the polarity of the composition. In short, Pandanus’ phytochemicals could be obtained via alcoholic extraction. The utilisation of modern chromatographic applications enables the scientists to discover and subsequently, purify single component of the complex natural mixtures. This finding can be beneficial to the research areas in botanical study, pharmacognostical assessment and biotechnology industry. It is aimed that more Pandanus species could be learnt for the future.

scholarly journals Analysis of Pandanus Extracts by Reversed Phase High Performance Liquid Chromatography

2016 ◽  
Vol 13 (2) ◽  
pp. 1
Author(s):  
Hannis Fadzillah Mohsin ◽  
Ratni Suriyani Jalal ◽  
Ibtisam Abdul Wahab ◽  
Abdul Rasyid Zulkifli ◽  
Muhammad Faxrurrazy Sadiran
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Ultra Violet ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Automated System ◽  
Elution Time ◽  
Research Areas ◽  
Alcoholic Extraction

The Pandanus species (Pandanaceae family) was investigated, in order to set up a library of their chromatographic profiles. From a literature review, the most common Pandanus plant that was greatly examined was P. amaryllifolius (the fragrant screw pine), followed by P. sanderi. Here, the chromatographic data from an extract of a relatively unfamiliar Pandanus is also presented. The leaves of P. monotheca were extracted by using methanol. Later, the extract was filtered, prior to a Reversed-Phase High Performance Liquid Chromatography (RP-HPLC) experiments. The automated system was set to run by gradient elution (acetonitrile:water). The flow rate was maintained at 1 ml/min, otherwise stated in a number of attempts. The ultra-violet absorbance of the Pandanus’ constituents was measured via Multiple Wavelength Detector. From the chromatogram, the major peaks for Pandanus’ compounds were eluted (λ = 220, 260 and 360 nm) and the retention times were recorded. The lower absorbance readings of other minor peaks could be due to various minor secondary metabolites, such as the furolignan. The different elution time depends on the polarity of the composition. In short, Pandanus’ phytochemicals could be obtained via alcoholic extraction. The utilisation of modern chromatographic applications enables the scientists to discover and subsequently, purify single component of the complex natural mixtures. This finding can be beneficial to the research areas in botanical study, pharmacognostical assessment and biotechnology industry. It is aimed that more Pandanus species could be learnt for the future. 

EVIDENCE FOR BIIIARY EXCRETION OF PHENPROCOUMON AND ITS METABOLITES IN HUMANS; IDENTIFICATION BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY AND GAS CHROMATOGRAPHY-MASS SPECTROMETRY

1987 ◽  
Author(s):  
J X de Vries ◽  
R Raedsch ◽  
A Stiehl ◽  
U Voelker ◽  
I Walter-Sack ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Gas Chromatography Mass Spectrometry ◽  
Phase Gradient ◽  
Chromatography Mass Spectrometry

Recently it has been shown that in man the oral couma-rin anticoagulant phenprocoumon is eliminated up to 60-70 % in urine and 30-40 % in faeces; in urine phenprocoumon (PH) and its metabolites 7-hydroxy-(7-OH),6-hydroxy-(6-OH) and 4'-hydroxy-(4'-OH) phenprocoumon are present mainly as conjugates. No data so far were available on the biliary excretion of these compounds.We examined bile obtained from four in-patients during PH treatment; bile samples were aspirated in the duodenum at the papilla during routine diagnostic endoscopy and immediately deep frozen before analysis. Samples were extracted both untreated as well as after hydrolysis with 6-glucuronidase/aryl sulfatase and separated by reversed phase gradient elution high performance liquid chromatography (HPLC) with fluorescence detection; for confirmation, the same extracts were methylated and analysed by gas chromatography-mass spectrometry (CG-MS) (J.X.de Vries et al J Chromatogr., 338 (1985) 325). PH, 7-OH, 6-OH and 4'-OH were identified by comparison with synthetic authentic samples'''''''

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