Quantitative Liquid Chromatographic Method Using Fluorescence Detection for Determining Zearalenone and Its Metabolites in Blood Plasma and Urine

1985 ◽  
Vol 68 (4) ◽  
pp. 632-635 ◽  
Author(s):  
Monica E Olsen ◽  
Hans I Pettersson ◽  
Kerstin A Sandholm ◽  
Karl-Heinz C Kiessling
Keyword(s):  
Blood Plasma ◽  
Mobile Phase ◽  
Chloroform Extract ◽  
Excitation Wavelength ◽  
Fluorescence Detector ◽  
Standard Ml ◽  
Blood And Urine Samples ◽  
Almost All ◽  
Water Saturated ◽  
Liquid Chromatographic

Abstract The liquid chromatographic (LC) method described, suitable for use with both blood plasma and urine, is applicable for determination of zearalenone and α-zearalenol at levels as low as 0.5 ng/mL plasma and 5 ng/mL urine. The sample is incubated overnight β with glucuronidase to analyze for both conjugated and unconjugated forms of zearalenone. The next day, the sample is acidified with H3P04, extracted with chloroform, and evaporated to dryness. The residue is dissolved in toluene and loaded onto a silica gel cartridge which is washed with toluene and eluted with toluene-acetone (88 + 12). The eluate is evaporated, and the residue is dissolved in chloroform, extracted with 0.18M NaOH, neutralized with H3PO4, and re-extracted with chloroform. The chloroform extract is evaporated, dissolved in mobile phase for LC, and injected onto a normal phase column under the following chromatographic conditions: mobile phase of water-saturated dichloromethane containing 2% 1-propanol, and fluorescence detector, excitation wavelength 236 nm, and 418 nm cut-off emission filter. Recoveries of zearalenone and its metabolites from blood plasma and urine are 80-89% in the range 2.0-10 ng standard/mL plasma, and 81-90% in the range 10-30 ng standard/mL urine. This method was used to analyze blood and urine samples from a pig fed zearalenone-contaminated feed (5 mg/kg), corresponding to 80 μg/kg body weight. Zearalenone was rapidly metabolized to a-zearalenol, which appeared in the blood only 30 min after feeding. Almost all zearalenone and αzearalenol was found conjugated with glucuronic acid in both blood plasma and urine.

High Performance Liquid Chromatographic Method Using Fluorescence Detection for Quantitative Analysis of Zearalenone and α-Zearalenol in Blood Plasma

1981 ◽  
Vol 64 (2) ◽  
pp. 302-310
Author(s):  
H Locksley Trenholm ◽  
Robert M Warner ◽  
Edward R Farnworth
Keyword(s):  
Blood Plasma ◽  
High Performance ◽  
Chromatographic Method ◽  
High Performance Liquid Chromatographic ◽  
Excitation Wavelength ◽  
Fluorescence Detector ◽  
Standard Ml ◽  
Liquid Chromatographic Method ◽  
Liquid Chromatographic ◽  
In Blood Plasma

Abstract A sensitive, high performance liquid chromatographic method is described for quantitative determination of zearalenone and α-zearalenol in blood plasma. Blood plasma is extracted with 2-propanol in ether, the extract is evaporated to dryness, and the residue is dissolved in 0.18N NaOH. The aqueous phase is washed with chloroform, dichloromethane, and benzene, neutralized with 0.10M H3PO4, and extracted with benzene. The extract is evaporated, dissolved in methanol, and injected onto a reverse phase column containing LiChrosorb RP-8 under the following conditions: methanol-acetonitrile-water mobile phase, fluorescence detector, excitation wavelength 236 nm, and 418 nm cut-off emission filter. The limit of detectability (twice background) is 0.5 ng standard which is equivalent to 0.6 ng standard/mL blood plasma. Linear standard curves are observed over the range of 0-35 ng of injected zearalenone and α-zearalenol. The recoveries from blood plasma are 76-101% in the range of 1.5-6.0 ng standard/mL blood.

Rapid, Sensitive Liquid Chromatographic Method for Determination of Zearalenone and α- and β-Zearalenol in Wheat

1984 ◽  
Vol 67 (5) ◽  
pp. 968-972
Author(s):  
H Locksley Trenholm ◽  
Robert M Warner ◽  
Dennis W Fitzpatrick
Keyword(s):  
Mobile Phase ◽  
Internal Standard ◽  
Automated Analysis ◽  
Excitation Wavelength ◽  
Uv Detector ◽  
Fluorescence Detector ◽  
Fluorescence Excitation ◽  
Final Residue ◽  
Liquid Chromatographic

Abstract A rapid, sensitive liquid chromatographic (LC) method is described for quantitative determination of zearalenone and α- and β-zearalenol in wheat. The procedure incorporates an internal standard, zearalenone oxime, to facilitate quantitation and automated analysis. A sample, buffered with pH 7.8 phosphate, is extracted with water-ethanolchloroform( 2 + 50 + 75) and cleaned up. The final residue is dissolved in LC mobile phase and injected onto a reverse phase RP-18 column under the following conditions: water-methanol-acetonitrile (5 + 3 + 2) mobile phase; fluorescence (excitation wavelength 236 nm, 418 nm cut-off emission filter) and UV (254 nm, range 0.0025 AU) detectors. The limit of detectability (twice background) is 0.5 ng for zearalenone and α-zearalenol standards on the fluorescence detector and 4 ng for β-zearaIenol on the UV detector, which is equivalent to 20 μg zearalenone and 20 μg α-zearalenol/kg, and 160 μg β-zearalenol/kg feed. Standard curves are linear over the range 0-35 ng zearalenone and α-zearalenol on the fluorescence detector and 0-50 ng β-zearalenol on the UV detector. Recoveries of all compounds are 87.5-101% in the range 0.1-3.0 mg/kg (ppm).

Liquid Chromatographic Determination of Ergot Alkaloids in Wheat

1986 ◽  
Vol 69 (4) ◽  
pp. 697-699
Author(s):  
George M Ware ◽  
Allen S Carman ◽  
Octave J Francis ◽  
Shia S Kuan
Keyword(s):  
Mobile Phase ◽  
Ergot Alkaloids ◽  
Ammonium Phosphate ◽  
Ammonium Hydroxide ◽  
Chromatographic Determination ◽  
Resin Column ◽  
Fluorescence Detector ◽  
Liquid Chromatographic Determination ◽  
Liquid Chromatographic

Abstract A method is described for the determination of individual ergot alkaloids in wheat. The sample is extracted with ethyl acetate-4% ammonium hydroxide (100 + 10), and the extract is cleaned up by liquidliquid partition. The ergot alkaloids are resolved by liquid chromatography (LC), using a porous cross-linked polystyrene-divinylbenzene resin column and a mobile phase consisting of acetonitrile-0.05M dibasic ammonium phosphate (55 + 45) buffered at pH 10.0. The ergot alkaloids ergonovine, ergonovinine, ergotamine, ergotaminine, α-ergocryptine, α-ergocryptinine, ergocristine, and ergocristinine are separated by LC and detected with a fluorescence detector. Recovery of ergot alkaloids added to wheat at levels of 16-760 ng/g averaged 85.6% with a coefficient of variation of 11.1%.

Improved liquid-chromatographic assay of labetalol in plasma.

1987 ◽  
Vol 33 (8) ◽  
pp. 1450-1452 ◽  
Author(s):  
D R Luke ◽  
G R Matzke ◽  
J T Clarkson ◽  
W M Awni
Keyword(s):  
High Performance Liquid Chromatography ◽  
Mobile Phase ◽  
High Performance ◽  
Internal Standard ◽  
Excitation Wavelength ◽  
Plasma Samples ◽  
Standard Curve ◽  
Low Concentrations ◽  
Styrene Divinylbenzene ◽  
Liquid Chromatographic

Abstract This is an assay for labetalol in plasma by "high-performance" liquid chromatography, with 5-(2-[4-(4-chlorophenyl)ethyl]) salicylamide hemihydrate as the internal standard. Plasma samples (500 microL) are extracted with acetonitrile, evaporated under nitrogen, reconstituted in the mobile phase, and injected onto a PRP-1 (Hamilton) column packed with particles of poly(styrene-divinylbenzene) copolymer. Fluorescence, enhanced by post-column introduction of NH4OH, was measured in the effluent (excitation wavelength 340 nm, emission wavelength 418 nm). Retention times for labetalol and the internal standard were 1.99 and 3.32 min, respectively. Inter- and intraday CVs for high and low concentrations of the drug were less than 7.5%. The assay standard curve is linear from 1 to 250 micrograms/L. Some commonly co-administered drugs were tested and did not interfere.

scholarly journals DETERMINATION OF LEVELS OF GLUCOSAMINE HYDROCHLORIDE AND CHONDROITIN SULFATE IN MIXTURES IN TABLET AND CREAM FORMS USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH FLUORESCENCE

2017 ◽  
Vol 9 ◽  
pp. 144
Author(s):  
Harmita Harmita ◽  
Catur Jatmika ◽  
Mufid Idan Nugraha
Keyword(s):  
Chondroitin Sulfate ◽  
Mobile Phase ◽  
High Performance ◽  
Bone Structure ◽  
Excitation Wavelength ◽  
Structural Components ◽  
Fluorescence Detector ◽  
Glucosamine Hydrochloride ◽  
Acetyl Group

Objective: Glucosamine hydrochloride (HCl) and chondroitin sulfate are glycosaminoglycan compounds and major structural components of bonesin the form of proteoglycans. These compounds maintain bone structure by stimulating the synthesis of synovial fluid and inhibiting the degradationof joint cartilage, and they can be used for the treatment of osteoarthritis. The aim of this study was to identify a selective analytical method fordetermining glucosamine HCl and chondroitin sulfate levels in tablet and cream forms.Methods: After derivatization using ortho-phthalaldehyde and 2-mercaptoethanol, the samples were analyzed using high-performance liquidchromatography (HPLC) with a fluorescence detector at an excitation wavelength of 335 nm and an emission wavelength of 445 nm. Deacetylationusing sodium hydroxide was required to break the acetyl group bond. The mobile phase used tetrahydrofuran 0.25% in water-acetonitrile (87:13)with a flow rate of 1.5 ml/minute.Results: The average levels of glucosamine HCl and chondroitin sulfate were 92.76% and 96.11% in tablets and 101.15% and 100.33% in creams,which fulfilled the acceptance criteria.Conclusions: Our validation method for glucosamine HCl and chondroitin sulfate met the acceptance criteria of accuracy, precision, selectivity, andlinearity.

Determination of urinary placental estriol by reversed-phase liquid chromatography with fluorescence detection.

1980 ◽  
Vol 26 (1) ◽  
pp. 130-132
Author(s):  
J T Taylor ◽  
J G Knotts ◽  
G J Schmidt
Keyword(s):  
Reversed Phase ◽  
Excitation Wavelength ◽  
Fluorescence Detector ◽  
Liquid Chromatograph ◽  
Chromatographic Procedure ◽  
Natural Fluorescence ◽  
Phase Liquid ◽  
Near Term ◽  
Far Ultraviolet ◽  
Liquid Chromatographic

Abstract We describe a liquid-chromatographic procedure for determining urinary estriol concentrations. The urine sample, after enzymatic hydrolysis to free the conjugated estrogen, is extracted with ether, and an aliquot of the resulting extraction residue is injected into the liquid chromatograph. Sample components are separated with a reversed-phase C18 column and isocratic elution with an acetonitrile/water mobile phase. Using a far-ultraviolet excitation wavelength, we measure the natural fluorescence of the eluted estrogen with a fluorescence detector. The procedure provides excellent sensitivity for determing near-term pregnancy concentrations of urinary estriol. The selectivity of the method limits the effect of potentially interfering compounds.

Liquid-chromatographic determination of cimetidine, its known metabolites, and creatinine in serum and urine.

1981 ◽  
Vol 27 (2) ◽  
pp. 272-275 ◽  
Author(s):  
J A Ziemniak ◽  
D A Chiarmonte ◽  
J J Schentag
Keyword(s):  
Mobile Phase ◽  
Methylene Chloride ◽  
Internal Standard ◽  
Ammonium Hydroxide ◽  
Isoamyl Alcohol ◽  
Chromatographic Determination ◽  
Water Saturated ◽  
Liquid Chromatographic ◽  
Serum And Urine

Abstract We describe a liquid-chromatographic procedure for determination of cimetidine, its hydroxymethyl-, sulfoxide-, and guanyl urea metabolites, and creatinine in patients serum and urine. SKF 92374 is used as the internal standard. Protein in 0.5 mL of serum or diluted urine is precipitated with 2 mL of acetonitrile, the organic and aqueous phases are separated by adding 0.3-0.5 g of anhydrous K2HPO4. The organic phase is evaporated, and 0.5 mL of 50 mmol/L HCl is added. This solution is washed with 3 mL of water-saturated isoamyl alcohol, the aqueous phase is extracted with 3 mL of methylene chloride and enough K2HPO4 to saturate the solution. The methylene chloride is evaporated, the residue reconstituted with 100 microL of mobile phase, and a 25-microL aliquot injected onto the chromatographic column (Dupont Sil). The mobile phase is acetonitrile/methanol/water/ammonium hydroxide (1000/50/50/2, by vol). The column effluent is monitored at 228 nm. Lower limits of detection ranged from 0.05 mg/L for cimetidine to 0.2 mg/L for guanyl urea. We determined cimetidine and its principal metabolites in the serum of a patient receiving cimetidine for the treatment of Zollinger-Ellison syndrome, and have assessed use of the assay in a clinical setting.

Determination of Ergotamine Tartrate in Tablets by Liquid Chromatography with Fluorescence Detection

1987 ◽  
Vol 70 (3) ◽  
pp. 538-540
Author(s):  
Ugo R Cieri
Keyword(s):  
Mobile Phase ◽  
Small Volume ◽  
Excitation Wavelength ◽  
Reverse Phase ◽  
Volumetric Flask ◽  
Relative Standard ◽  
Liquid Chromatographic ◽  
Ergotamine Tartrate ◽  
Phase Column

Abstract A method is presented for the liquid chromatographic (LC) determination of ergotamine tartrate in tablets that is applicable even in the presence of other ingredients such as phenobarbital, belladonna alkaloids, and caffeine. The sample is transferred to a volumetric flask, a small volume of formic acid is added to dissolve and stabilize the ergotamine, and the solution is diluted to volume with methanol. The solution is mixed and filtered through paper. The LC system consists of a Rheodyne injector fitted with a 20 μL loop and a C,18 reverse phase column; the mobile phase is acetonitrile-water-triethylamine (700 + 300 + 0.5). Ergotamine tartrate is determined fluorometrically at an excitation wavelength of 250 nm and an emission wavelength of 430 nm. Recovery studies were conducted at the 0.3 and 1.0 mg levels. Average recoveries were 99.6 and 100.8%, respectively; relative standard deviations (RSDs) were 1.08 and 2.21%, respectively. Some commercial preparations containing ergotamine tartrate in combination with other ingredients were also analyzed. The RSDs for 5 determinations of each of 2 ground composites were 0.09 and 0.34%.

High Performance Liquid Chromatographic Determination of Aflatoxicol in Milk, Blood, and Liver

1981 ◽  
Vol 64 (5) ◽  
pp. 1083-1087 ◽  
Author(s):  
Mary W Trucksess ◽  
Leonard Stoloff
Keyword(s):  
Mobile Phase ◽  
High Performance ◽  
Raw Milk ◽  
Chromatographic Determination ◽  
High Performance Liquid Chromatographic ◽  
Fluorescence Detector ◽  
Beef Liver ◽  
Liquid Chromatographic Determination ◽  
Liquid Chromatographic

Abstract Samples of milk were extracted with chloroform, the extract was transferred to methanol, and residual interferences were removed by liquid-liquid partition against hexane and by silica gel column chromatography. Aflatoxicol (AFL) in the purified extract was resolved on a μBondapak C18 column, using water - methanol - acetonitrile - tetrahydrofuran (70 + 15 + 20 + 3) as the mobile phase, and measured with a fluorescence detector (excitation 325-385 nm, emission >408 nm). Recoveries of AFL added to samples of whole pasteurized milk at levels ranging from 0.025 to 0.10 ng/mL averaged 92% (range 78- 100%). The method for AFL has also been applied to the analysis of raw milk, whole beef blood, and beef liver, with recoveries of 70-88%.

Determination of urinary placental estriol by reversed-phase liquid chromatography with fluorescence detection.

1980 ◽  
Vol 26 (1) ◽  
pp. 130-132 ◽  
Author(s):  
J T Taylor ◽  
J G Knotts ◽  
G J Schmidt
Keyword(s):  
Reversed Phase ◽  
Excitation Wavelength ◽  
Fluorescence Detector ◽  
Liquid Chromatograph ◽  
Chromatographic Procedure ◽  
Natural Fluorescence ◽  
Phase Liquid ◽  
Near Term ◽  
Far Ultraviolet ◽  
Liquid Chromatographic

Abstract We describe a liquid-chromatographic procedure for determining urinary estriol concentrations. The urine sample, after enzymatic hydrolysis to free the conjugated estrogen, is extracted with ether, and an aliquot of the resulting extraction residue is injected into the liquid chromatograph. Sample components are separated with a reversed-phase C18 column and isocratic elution with an acetonitrile/water mobile phase. Using a far-ultraviolet excitation wavelength, we measure the natural fluorescence of the eluted estrogen with a fluorescence detector. The procedure provides excellent sensitivity for determing near-term pregnancy concentrations of urinary estriol. The selectivity of the method limits the effect of potentially interfering compounds.

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