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1.
A HPLC method with UV/diode array detection for the determination of deoxynivalenol (DON) and deepoxy-deoxynivalenol (DOM-1) in milk was developed. Milk was incubated with β-glucuronidase and then defatted. After purification by immunoaffinity chromatography, DON and DOM-1 were separated on a C18 reversed phase column with acetonitril/water (10/90) as the mobile phase and detected at 218 nm. Limits of quantification were 1 μg/l for both toxins, with mean recoveries (1–10 μg/l) of 97% (DON) and 84% (DOM-1), respectively. Milk samples (pasteurized, UHT; n=32) from German retail shops were analysed by this method. Neither DON/DOM-1 nor their glucuronides were found in any sample. These results are consistent with published studies indicating that in lactating cows, DON and DOM-1 are mostly eliminated through urine, and that the carry-over into milk is negligible. 相似文献
2.
Carry-over of deoxynivalenol (DON) into eggs was investigated within the scope of a 16-week experiment with laying hens, in
which the birds were fed a maize-based diet containing DON at 11.9 mg/kg dry matter. Eggs were collected during weeks 2, 4,
8, and 16. DON and its metabolite deepoxy-DON were analysed separately in freeze-dried yolk and albumen. Yolk was extracted
with water and the extract was purified using an immunoaffinity column (IAC). Albumen was extracted with acetonitrile-water
and the extract was pre-cleaned before applying an IAC. All albumen and some yolk samples were incubated with β-glucuronidase
prior to extraction. DON and de-epoxy-DON were determined by high performance liquid chromatography (HPLC) with diode array
detection (DAD). The detection limits of both toxins were 20 ng/g and 15 ng/g in freezedried yolk and albumen, respectively,
corresponding to approximately 10 ng/g and 2 ng/g in fresh samples. The recovery of DON/de-epoxy-DON in spiked samples (50–200
ng/g) was 87/83% (yolk) and 87/77% (albumen) with coefficients of variation of 4–15%. Neither DON nor de-epoxy-DON were detected
in any of the samples. In order to achieve lower detection limits, the methods are currently optimized. However, these preliminary
results indicate that eggs do not contribute significantly to the dietary DON intake of the consumer.
Presented at the 26th Mykotoxin-Workshop in Herrsching, Germany, May 17–19, 2004 相似文献
3.
Methods to determine zearalenone (ZEA), deoxynivalenol (DON), aflatoxins (AF) and their metabolites in pig urine were developed
as biomarkers for pig exposure to the mycotoxins in feed. Urine samples were incubated with β-glucuronidase to cleave conjugates,
extracted and cleaned-up with solid phase and immunoaffinity columns, followed by HPLC with UV and fluorescence detection.
Good recoveries (83–130%), low variation (2–10%), and low detection limits (0.3–9.9 ng/ml) were obtained. The results of controlled
AFB1 feeding trials found no difference in urine concentrations of AFB1 or AFM1 from pigs fed three different levels (127, 227, 327 μg/kg) of AFB1 in diets. The excretion of AFB1 and AFM1 in urine was on average 30% of the oral dose and the ratio AFB1 to AFM1 was around 23%. The analysis of 15 Vietnamese pig urine samples indicate a relatively high exposure of ZEA, DON and AF, which
were found as toxin or metabolites in 47, 73, and 80% of the urine samples, respectively. 相似文献
4.
Concentrations of deoxynivalenol (DON) and deepoxy deoxynivalenol (DOM-1) in animal blood are important parameters for studies
in toxicology and biological detoxification of DON. Clean-up methods, using either immunoaffinity chromatography (IAC) or
solid phase extraction (SPE), were compared in order to determine the free form of DON or DOM-1 and the sum amount (free form
plus glucuronide conjugated form of DON or DOM-1), respectively, in swine serum. Detection was achieved by high performance
liquid chromatography with ultraviolet detection (HPLC-UV). Compared with the SPE-HPLC method, the IAC-HPLC method provided
lower quantitation limit (DON: 18 vs 42 ng/ml; DOM-1: 21 vs 30 ng/ml) and higher recoveries (DON: 93.4–102.7% vs 63.7–85.3%;
DOM-1: 85.5–91.1% vs 68.0–82.6%). Compared with previously published methods, the developed IAC-HPLC method removed analytical
interferences from swine serum in one quick and easy step, and eliminated steps of extraction with organic solvent and/or
pre-purification using SPE cartridges. This IAC-HPLC method was used to analyze swine serum samples collected from pigs that
were evaluated in a feeding trial of a microbiological detoxification of DON. No DON or DOM-1 were detected in serum samples
from pigs given a toxin-free diet or a microbial control diet. In serum samples from pigs given a DON diet (5 mg/kg of DON),
free form DON and sum free DON + conjugated DON were 38.8 ± 13.7 and 49.8 ± 14.1 ng/ml, respectively. In serum samples from
those given a detoxified-DON diet (DON was transformed to DOM-1), free form DOM-1 was detected but not quantified, and the
sum DOM-1 was found as 47.5 ± 6.3 ng/ml. 相似文献
5.
Simple method for isolation of 4-deoxynivalenol from rice inoculated with Fusarium graminearum. 总被引:5,自引:5,他引:0 下载免费PDF全文
A new method for preparative isolation of 4-deoxynivalenol (DON) is presented. This method avoids the loss of material during purification on silica gel by column chromatography. DON and 3-acetyldeoxynivalenol in crude extracts of rice inoculated with Fusarium graminearum were converted to triacetyldeoxynivalenol; the acetylated product was easier to purify by silica gel chromatography than DON is. After hydrolysis and further purification on a charcoal-alumina column, the 71% pure DON was recovered in yields as high as 450 mg of DON per kg of rice. Subsequent separation on a Sephadex LH20 column yielded DON that was greater than 90% pure. 相似文献
6.
Fusarium toxins are secondary metabolites produced byfungi of these genera in many commodities under certain conditions. A study was carried out to investigate the co-occurrence of
zearalenone (ZEN), deoxynivalenol (DON) and fumonisins (FB1 and FB2) in 52 samples of mixed-feed for poultry contaminated withFusarium verticillioides. The zearalenone and deoxynivalenol were checked using immunoaffinity column and the extraction of fumonisin was performed
by strong anion exchange (SAX) solid phase column. Detection and quantification were determined by high performance liquid
chromatography (HPLC). The limit of detection was 5 μg/kg for ZEN, 100 μg/kg for DON and 50 and 100 μg/kg for FB1 and FB2 respectively.Fusarium toxins were detected in 20 samples. Sixteen samples were positive for ZEN (30.7%) presenting levels that ranged from 7.4
μg/kg to 61.4 μg/kg (mean=27.0 μg/kg). 13.5% of the samples presented contaminations of DON, with levels ranging from 100.0
μg/kg to 253 μg/kg (mean=l18.07 μg/kg). FB1 was detected in 19.2% of samples, with levels ranging from 50.0 μg/kg to 110.0 μg/kg (mean=73.6 μg/kg). FB2 was not detected
in any sample. In positive samples simultaneously contamination with two or three mycotoxins were detected in 9 of them (17.3%). 相似文献
7.
Adsorption characteristics of an anti-peptide antibody, obtained by immunization of eight amino acids in the C-terminal region
of chimeric α-amylase of rice α-amylase isozymes, were studied by use of the chimeric enzyme and the peptide used for immunization.
This anti-peptide antibody adsorbed the enzyme, as well as the peptide antigen, with sufficient affinity for immunoaffinity
purification and was used for purification of the enzyme secreted from yeast cells. Chimeric α-amylase was purified by immunoaffinity
chromatography to high purity in one step from the fermentation broth. One-third of the secreted enzyme was not adsorbed by
the column of anti-peptide antibody because of processing in the C-terminal region. 相似文献
8.
A total of 92 samples — 23 winter wheat, 12 summer barley, 5 oats and 52 mixed feed — were collected from a state factory
in Kaunas, Lithuania and were analysed for the presence of trichothecenes, zearalenone (ZEN) and ochratoxin A (OA) using gas
chromatography with electron capture detection and immunoaffinity column/high performance liquid chromatography with fluorescence
and UV detections. Deoxynivalenol (DON), nivalenol (NIV), T-2 toxin and HT-2 toxin were detected at concentrations above 10
μg/kg in 68%, 48%, 38% and 8% of cereal samples, respectively, and in 98%, 88%, 12% and 8% of samples of mixed feed for swine
and poultry. More than 10 μg/kg of zearalenone and ochratoxin A were found in 58% and 92% of the mixed feed samples, respectively.
The highest concentrations of all analysed trichothecenes in Lithuanian mixed feed and cereal grains, with an exception of
T-2 toxin in one oat lot and one sample of mixed feed and OA in two mixed feed samples, were lower than those reported as
Lithuanian advisory or tolerance limits. 相似文献
9.
Physiological samples of 100 piglets fed diets containing 0.01, 0.06, 0.15, 0.22 and 0.42 mg ZON and 0.2, 0.8, 1.0, 1.9 and
3.9 mg DON per kg over a period of 35±1.5 days were investigated for concentrations of deoxynivalenol (DON) and zearalenone
(ZON) and their metabolites.
DON was detected in serum, bile and urine in increasing concentrations corresponding to the diet contamination. The metabolite
de-epoxy-DON was detected only in urine. The DON contamination of the diet was closely reflected by the serum concentrations
of the piglets.
ZON and its metabolite α-zearalenol were detected in bile fluid, liver and urine, while β-zearalenol was only detected in
bile fluid. In serum neither ZON nor its metabolites were found. The total concentration of ZON plus its metabolites in the
bile fluid corresponded well with the dietary contamination.
For all analyses it has to be noted that toxin residues were detectable even in individual samples of piglets fed the control
diet containing 0.01 mg ZON/kg and 0.2 mg DON/kg.
Presented at the 25th Mykotoxin Workshop in Giessen, Germany, May 19–21, 2003 相似文献
10.
We have purified luteinizing hormone-releasing hormone (LH-RH) from codfish brain and have demonstrated its identity with salmon LH-RH (sLH-RH). An antiserum raised against sLH-RH was used in a specific radioimmunoassay (RIA) to monitor purification and to manufacture an immunoaffinity chromatography column for the initial purification step. The cross-reactivity of the sLH-RH RIA with mammalian LH-RH was 0.1%. Acid extracts of codfish brains were sequentially purified by immunoaffinity chromatography, gel-filtration chromatography, and three steps of reverse-phase HPLC. The purified material and synthetic sLH-RH coeluted on reverse-phase HPLC and exhibited similar biological activity in a dispersed pituitary cell bioassay. Furthermore, the amino acid composition of the purified material was identical to salmon LH-RH. These results suggest that there is structural conservation of LH-RH between these species of teleost fish. 相似文献
11.
A simple method for determination of deoxynivalenol (DON) in cereal samples is described. DON was extracted with methanol,
the solvent evaporated, and the residue redissolved with water. This extract was purified on immunoaffinity columns. DON was
determined by HPLC with UV-detection. The limits of detection (LOD) and quantification (LOQ) were 10 and 50 μg/kg, respectively.
Presented at the 25th Mykotoxin Workshop in Giessen, Germany, May 19–21, 2003 相似文献
12.
Changes in the Levels of Abscisic Acid and Its Metabolites in Excised Leaf Blades of Xanthium strumarium during and after Water Stress 总被引:15,自引:11,他引:4 下载免费PDF全文
Zeevaart JA 《Plant physiology》1980,66(4):672-678
The time course of abscisic acid (ABA) accumulation during water stress and of degradation following rehydration was investigated by analyzing the levels of ABA and its metabolites phaseic acid (PA) and alkalihydrolyzable conjugated ABA in excised leaf blades of Xanthium strumarium. Initial purification was by reverse-phase, preparative, high performance liquid chromatography (HPLC) which did not require prior partitioning. ABA and PA were purified further by analytical HPLC with a μBondapak-NH2 column, and quantified by GLC with an electron capture detector. 相似文献
13.
Li Y Wang Z De Saeger S Shi W Li C Zhang S Cao X Shen J 《Methods (San Diego, Calif.)》2012,56(2):192-197
An immunoaffinity column (IAC) was prepared with a new deoxynivalenol (DON) monoclonal antibody and used as a clean-up tool before ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) analysis of DON in cereals. The developed IAC clean-up method showed high recoveries for DON. They ranged from 61% to 103% in wheat, rice, and millet with intra-day and inter-day variations below 19% and 17%, respectively. The column capacity was 2.86μg DON per mL of gel, and it maintained above 0.68μg/mL of gel after 10 cycles of usage at 2 days intervals. The limit of detection (LOD) and limit of quantification (LOQ) were 0.3 and 0.8μg/kg, respectively. Twenty-one out of 40 analyzed commercial cereal samples were positive at DON concentrations from 7 to 534μg/kg. 相似文献
14.
Efficient large-scale purification of restriction fragments by solute-displacement ion-exchange HPLC. 下载免费PDF全文
Extreme overloading of HPLC columns with sample can create a condition of binding site saturation causing competition and displacement among solutes during column elution. This has been termed solute-displacement chromatography (SD-HPLC). We present an example of this phenomenon for the preparative fractionation and purification of restriction fragments of almost identical size (1337 and 1388 bp) which cannot be resolved by agarose gel electrophoresis. Standard analytical ion-exchange HPLC chromatography failed to separate these fragments from each other and from an unexpectedly early eluting pUC-derived vector fragment of 2.7 kbp. We demonstrate that by intentional overloading of the small (4.6 x 35 mm) non-porous TSK-DEAE HPLC column, hundreds of micrograms of DNA restriction fragments could be resolved and purified in a single HPLC run of less than 30 minutes. 相似文献
15.
Ulvskov Peter; Marcussen Jan; Rajagopal Ranganatha; Prinsen Els; Rudelsheim Patrick; Onckelen Henri Van 《Plant & cell physiology》1987,28(5):937-945
A single-step immunoaffinity purification procedure using monoclonalantibodies was developed to isolate indole-3-acetamide fromplant extracts. Antibodies from a selected clone, raised againstIAA-Cl'-BSA, with pronounced ability to recognize indole-3-acetamide(IAM) were used to prepare an immunoaffinity absorbent. Antibodiespurified by thiophilic interaction chromatography were immobilizedon divinylsulfoneactivated agarose. This column shows a veryhigh selectivity towards IAM compared to IAA. This single stepof immunoaffinity purification gave plant extracts of sufficientpurity for direct quantification by on-line spectrofluorimetryafter an analytical ionsuppression-HPLC run. Successive approximationby a second analytical ion-pairing-HPLC run confirmed the validityof this analytical technique. (Received November 15, 1986; Accepted May 7, 1987) 相似文献
16.
Josef Böhm Lisa Koinig Ebrahim Razzazi-Fazeli Anja Blajet-Kosicka Magda Twaruzek Jan Grajewski Christiane Lang 《Mycotoxin Research》2010,26(3):147-153
The aim of the present study was to investigate the occurrence of mycotoxins in commercial dog food, as a basis to estimate
the risk of adverse effects. Seventy-six dry dog food samples from 27 producers were purchased from retail shops, supermarkets,
and specialized pet food shops in Vienna, Austria. The frequency and levels of deoxynivalenol (DON), zearalenone (ZEA), fumonisins
(FUM), ochratoxin A (OTA). and aflatoxins (AF) in dry dog food were determined. Mycotoxin analysis were performed by commercial
enzyme-linked immunosorbent assay (ELISA) test kits. Confirmatory analyses were done for DON, ZEA, and FUM by high performance
liquid chromatography (HPLC) after extract clean-up with immunoaffinity columns. The correlations between ELISA and HPLC results
for DON and ZEA were acceptable and indicated that ELISA could be a simple, low cost, and sensitive screening tool for mycotoxins
detection, contributing to quality and safety of pet food. DON was the mycotoxin most frequently found (83% positives; median
308 μg/kg, maximum 1,390 μg/kg). ZEA (47% positives, median 51 μg/kg and maximum 298 μg/kg) and FUM (42% positives, median
122 μg/kg and maximum 568 μg/kg) were also frequently detected in dog food. OTA was less frequently found (5%, median 3.6 μg/kg,
maximum 4.7 μg/kg. AF were not detected (<0.5 μg/kg) in any sample. The results show that dry dog food marketed in Vienna
are frequently contaminated with mycotoxins (DON > ZEA > FUM > OTA) in low concentrations, but do not contain AF. The high
frequency of Fusarium toxins DON, ZEA, and FUM indicates the need for intensive control measures to prevent mycotoxins in dog foods. The mycotoxin
levels found in dry dog food are considered as safe in aspects of acute mycotoxicoses. However, repeated and long-time exposure
of dogs to low levels of mycotoxins may pose a health risk. 相似文献
17.
Cherukury Madhu Zoltan Gregus Curtis D. Klaassen 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1995,674(2)
A simple HPLC method has been described to quantify diquat in biological fluids and tissues. This method permits separation and quantification of diquat from blood, bile, urine, liver and kidney. It does not require special pretreatment of the samples prior to analysis, nor a specially prepared analytical column. Various concentrations of diquat were added (10–300 nmol/ml or g) to fluids or tissues. Analysis of blank samples revealed no substrates that interfere with diquat elution. Excellent recovery (95–105%) was obtained. Diquat (120 μmol/kg, i.v.) was injected to rats and quantified in bile, blood and liver. Concentration of diquat was higher in blood and bile than liver. Therefore, this method is applicable for quantification of diquat in toxicological samples, and may be used to determine structurally similar compounds such as paraquat. 相似文献
18.
Inactive renin was purified to apparent homogeneity from human plasma by ion exchange, gel filtration, Affi-Gel blue, immunoaffinity chromatography on profragment-specific IgG coupled to Sepharose, and preparative HPLC. By this method, a 460000-fold purification was obtained. The purified renin was totally inactive and was activated by trypsin. 相似文献
19.
Minsun Baek Young-Soo Rho Dong-Hyun Kim 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1999,732(2):95
A column-switching high-performance liquid chromatography (HPLC) method is described for the determination of asiaticoside in rat plasma and bile using column-switching and ultraviolet (UV) absorbance detection. Plasma was simply deproteinated with acetonitrile prior to injection and bile was directly injected onto the HPLC system consisting of a clean-up column, a concentrating column, and an analytical column, which were connected with two six-port switching valves. Detection of asiaticoside was accurate and repeatable, with a limit of quantification of 0.125 μg/ml in plasma and 1 μg/ml in bile. The calibration curves were linear in a concentration range of 0.125–2.5 μg/ml and 1–20 μg/ml for asiaticoside in rat plasma and bile, respectively. This method has been successfully applied to determine the level of asiaticoside in rat plasma and bile samples from pharmacokinetics and biliary excretion studies. 相似文献
20.
Determination of zearalenone and its metabolites in urine, plasma and faeces of horses by HPLC-APCI-MS 总被引:5,自引:0,他引:5
Songsermsakul P Sontag G Cichna-Markl M Zentek J Razzazi-Fazeli E 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2006,843(2):252-261
The paper describes a method for the sensitive and selective determination of zearalenone and its metabolites in urine, plasma and faeces of horses by high performance liquid chromatography and atmospheric pressure chemical ionisation (APCI) mass spectrometry (MS). While only one step sample clean-up by an immunoaffinity column (IAC) was sufficient for plasma samples, urine and faeces samples had to be prepared by a combination of a solid-phase extraction (SPE) and an immunoaffinity column. The method allows the simultaneous determination of zearalenone and all of its metabolites; alpha-zearalenol, beta-zearalenol, alpha-zearalanol, beta-zearalanol and zearalanone. Dideuterated zearalanone was used as internal standard for quantification and the study of the matrix effect. Recovery rates between 56 and slightly above 100% were achieved in urine samples, and more than 80% in plasma and faeces samples. The limits of detection ranged from 0.1-0.5 microg/l or microg/kg, the limits of quantification from 0.5-1.0 microg/l or microg/kg. The practical use of the method is demonstrated by the analysis of spiked and naturally contaminated urine, plasma and faeces of horses. 相似文献