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1.
Maillard products, such as Nepsilon-carboxymethyllysine (CML), are readily formed during the manufacturing of infant formulas. Little has been known, however, about the presence of CML in human breast milk and about the uptake of CML by infants. In this study, CML was measured in the serum and breast milk of 32 healthy mothers by ELISA. CML concentrations in breast milk (137 +/- 82.7 ng/mL) were significantly lower than in the serum (399 +/- 67.8 ng/mL, p < 0.001) and on average 35-fold lower than in infant formulas (4754 +/- 4299.5 ng/mL). CML was also measured in the urine of 21 infants, which were fed with breast milk or formulas. Although there was a tendency toward higher urinary CML excretion in infants fed with hypoallergenic formulas compared to breast-fed ones, the differences were not significant. Neonates that were delivered by vaginal birth had significantly higher concentrations of CML compared to those delivered by caesarean section (1306 +/- 653 vs 601 +/- 220 ng/mL, p = 0.012). It is concluded that CML passes from the serum into the breast milk, but the levels are by far lower than in infant formulas. In very young neonates (< or =3 days), the mode of delivery has a greater influence on urinary CML excretion than the nutrition.  相似文献   

2.
A liquid chromatographic (LC) method is described for the determination of the plant estrogens diadzein, formononetin, and coumestrol and the estrogenically active metabolite equol in bovine blood plasma and urine. The blood and urine samples are incubated overnight with and without beta-glucuronidase/sulfatase for analysis of both free and conjugated forms of estrogens. Samples are applied to Extrelut columns, extracted with ethyl acetate, and evaporated to dryness. Residues from urine samples are dissolved in methanol, diluted with water, acidified with HCl, and purified by injection through a Sep-Pak C18 cartridge. This eluate is used for LC analysis. Residues from blood samples are dissolved in benzene-petroleum ether (1 + 1), extracted with ammonium hydroxide, acidified with glacial acetic acid, and extracted with ethyl acetate. The ethyl acetate extract is evaporated, dissolved in 80% methanol, injected onto a LC reverse-phase column, and separated in a linear gradient system between 40 and 80% methanol in phosphate buffer. Quantitation is performed by means of UV and fluorescence responses. The method was sensitive enough to determine 0.4 ng/mL of daidzein and formononetin and 0.1 and 13 ng/mL of coumestrol and equol, respectively, in blood, and 130, 80, and 7 ng/mL of daidzein, formononetin, and coumestrol, respectively, and 4 micrograms/mL of equol in urine. The applicability of the method was checked by the determination of total and free plant estrogens in blood samples from a dairy cow fed a normal diet.  相似文献   

3.
In this study, the residue depletion of nitrovin in chicken was studied after feeding the birds with dietary feeds containing 10 mg/kg of nitrovin for 7 consecutive days. Tissues (muscle, fat, kidney, and liver) and plasma were collected at different withdrawal periods and determined by a high-performance liquid chromatography-ultraviolet (HPLC-UV) method. The limit of detection for nitrovin in tissue and plasma samples was 0.1 ng/(g or mL), and the inter- and intrarecoveries from the blank fortified samples were in the range of 71.1-85.7%. At the withdrawal period of 0 days, the residue concentration of nitrovin in plasma was the highest (average of 84.98 ng/mL) compared to those in muscle, fat, liver, and kidney (average of 21.04, 61.18, 24.04, and 68.28 ng/g, respectively). At the withdrawal period of 28 days, the residue levels of nitrovin in muscle, fat, liver, and plasma were all higher than 1.0 ng/(g or mL) and the highest concentration was in liver (average of 5.8 ng/g). These data are in support of the ban of nitrovin as a feed additive in food-producing animals.  相似文献   

4.
A monoclonal antibody-based ractopamine immunoassay has been applied to incurred samples from sheep and cattle. Results obtained by immunoassay were compared with those from high-performance liquid chromatography (HPLC). Three sets of sample extracts containing primarily unmetabolized ractopamine were analyzed. Correlation of HPLC with enzyme-linked immunosorbent assay (ELISA) for beef liver samples gave an r(2) = 0.98 despite rather low ractopamine concentrations (range 1.1-13.4 ng/mL, n = 6). Ractopamine concentrations in cow urine samples treated by solid phase extraction, to remove ractopamine metabolites, also showed a high correlation between the HPLC and the ELISA results (r(2) = 0.95, range 1.0-275 ng/mL, n = 61). In contrast, HPLC and ELISA analyses of ractopamine in sheep urine were not well-correlated (r(2) = 0.58, range 0.85-51 ng/mL, n = 34). When ractopamine conjugates in urine samples were hydrolyzed with hydrolytic enzymes, ELISA and HPLC methods were highly correlated [r(2) = 0.94 for sheep (range 123-10 554 ppb, n = 60) and an r(2) = 0.98 for cattle (range 14-8159 ppb, n = 62)]. Tissues contained only minute amounts of ractopamine, and after 7-day withdrawal periods, less than 1 ppb of free ractopamine was detected. Ractopamine was rapidly metabolized in both cattle and sheep. The difference in ractopamine concentration of urine samples before and after hydrolysis indicated that only 1-5% of ractopamine was excreted unmetabolized. Results from this study indicate that the monoclonal antibody-based ELISA could be useful for a sensitive, quantitative, or qualitative ractopamine screening assay.  相似文献   

5.
Ractopamine hydrochloride is a beta-adrenergic leanness-enhancing agent approved for use in swine in the United States. Depletion of ractopamine and its metabolites from animal tissues, urine, and serum is of interest for the detection of illegal use. The objectives of this study were to measure the residues of ractopamine in swine incurred samples after treatment with dietary ractopamine for 28 consecutive days. An efficient and sensitive analytical method was developed for the detection of parent ractopamine and its metabolites in swine tissues, urine, and serum by HPLC-FLD. After extraction, enzymatic digestion, and solid-phase cleanup of the samples, ractopamine residues were determined by liquid chromatography (LC) with fluorescence detector. The limits of detection (LOD) for tissues, urine, and serum were 1 ng g(-1), 0.5 ng mL(-1), and 0.5 ng mL(-1), respectively. Recoveries ranged from 70.5 to 94.5% for samples fortified at 1-50 ng g(-1) or ng mL(-1). Sixty pigs were fed twice daily for 28 consecutive days with feeds containing 18 mg kg(-1) ractopamine HCl. The residue concentrations in urine, liver, and kidney were 650.06 ng mL(-1), 46.09 ng g(-1), and 169.27 ng g(-1), respectively, compared with those in muscle, fat, and serum (4.94 ng g(-1), 3.28 ng g(-1), and 7.48 ng mL(-1), respectively) at the feeding period of 7 days. The residue concentrations at withdrawal period of 0 days in all edible tissues were lower than tolerance values established by the FDA and MRL values listed by the JECFA. These data support the withdrawal time of 0 days established by the FDA for ractopamine used as feed additive in swine.  相似文献   

6.
Perchlorate exposure and potential effects were evaluated in large mammals by monitoring heifer calves placed on a site with access to streamwater fed by a perchlorate-contaminated groundwater spring ( approximately 25 ng/mL). Blood was collected from the two calves on the site (and two control calves from an uncontaminated site) approximately every 2 weeks for analysis of perchlorate residues and thyroid hormones. During the 14 week study, perchlorate was detected (detection limit = 13.7 ng/mL) in blood plasma twice (15 ng/mL and 22 ng/mL) in one of the heifer calves drinking perchlorate-contaminated water on consecutive sampling periods 4 and 6 weeks after the beginning of perchlorate exposure. Constant exposure to 25 ppb perchlorate in drinking water had no effect on circulating thyroid hormones (T(3) and T(4)) in the heifer calves.  相似文献   

7.
4-chloro-androst-4-ene-3,17-dione (CLAD) and 4-chlorotestosterone (clostebol, beta-CLT or CLT) were made immunogenic by coupling to protein carriers via the 3 and 17 positions, respectively. These immunogens were used to elicit polyclonal and monoclonal antibodies to CLAD and to clostebol. The antibodies were characterized in an enzyme immunoassay for sensitivity and specificity. Polyclonal antisera generated through position 17 reacted preferentially with 4-chlorotestosterone-17-acetate (clostebol acetate, CLTA), 4-chloro-epitestosterone (epi-clostebol, 17alpha-clostebol, 17alpha-CLT), and clostebol, whereas polyclonal antisera generated through the 3 position almost did not react with these derivatives. Interestingly, the monoclonal antibody generated through the 3 position recognized (35%) epi-clostebol. These results suggest that polyclonal antisera generated through the 17 position have a broad specificity profile and can be used to analyze by immunoassay methods urinary metabolites of clostebol acetate and thereby detect the illegal use of clostebol acetate in livestock farming.  相似文献   

8.
The glycosides of flavonoid, anthocyanins and A type proanthocyanidins in cranberry concentrate were characterized and quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Cranberry concentrate (1 g/body weight) was orally gavaged to Fischer-344 rats (n = 6), and blood and urine samples were collected over 24 h periods. Quercetin, 3'-O-methylquercetin (isorhamnetin), myricetin, kaempferol, and proanthocyanidin dimer A2, together with thirteen conjugated metabolites of quercetin and methylquercetin and intact peonidin 3-O-galactoside and cyanidin 3-O-galactoside were identified in the rat urine after cranberry treatment. Very low levels of isorhamnetin (0.48 ± 0.09 ng/mL) and proanthocyanidin dimer A2 (0.541 ± 0.10 ng/mL) were found in plasma samples after 1 h of cranberry administration. Although no quercetin was detected in plasma, MRM analysis of the methanolic extract of urinary bladder showed that chronic administration of cranberry concentrate to rats resulted in accumulation of quercetin and isorhamnetin in the bladder. These results demonstrate that cranberry components undergo rapid metabolism and elimination into the urine of rats and are present in the urinary bladder tissue potentially allowing them to inhibit urinary bladder carcinogenesis.  相似文献   

9.
To develop an enzyme-linked immunosorbent assay for the fungicide fenarimol, two synthesized haptens, haptens-1 and -2, and the purchased 4,4'-DDA were conjugated to carrier proteins (BSA, KLH, and OVA). Polyclonal antibodies raised against hapten-1,2-KLH conjugates in rabbits and the coating antigens of hapten-1,2-BSA conjugates, hapten-2-OVA conjugate, and 4,4'-DDA-BSA conjugate were screened and selected for the homologous and/or heterologous ELISA formats. Two competitive indirect ELISAs were selected: assays I and II. The optimized ciELISAs of assays I and II showed average IC(50) values of fenarimol of 5.4 and 9.4 ng/mL, detection ranges of 1.1-25.9 and 1.1-82.7 ng/mL, and lowest detection limits of 0.3 and 0.3 ng/mL, respectively. The cross-reactivities with several structurally related compounds indicated the importance of the steric fitness in the antigen-antibody interaction. Recoveries of fenarimol from apple and pear samples spiked with the analyte by assay I were in the range of 93-113% by simple extraction, concentration, and dilution. This assay could be a convenient and supplemental analytical tool for monitoring fenarimol residues in environmental and agricultural samples.  相似文献   

10.
gamma-Terpinene was mixed in artificial diet at a concentration of 1 mg/g of diet, and the diet was fed to the last instar larvae of common cutworm (Spodoptera litura). Metabolites were recovered from frass and analyzed spectroscopically. gamma-Terpinene was transformed mainly to p-mentha-1,4-dien-7-oic acid and p-cymen-7-oic acid (cumic acid). Similarly, (-)-alpha-phellandrene was transformed mainly to (4R)-p-mentha-1,5-dien-7-oic acid and p-cymen-7-oic acid (cumic acid). The C-7 position (allylic methyl group) of gamma-terpinene and (-)-alpha-phellandrene was preferentially oxidized.  相似文献   

11.
A simple method was developed for the determination of free and/or total isoflavones daidzein, genistein, and their respective 4'-methoxy derivatives biochanin A and formononetin (biochanin B) at low levels in human urine. A solid-phase extraction on octadecyl silica (C(18)) columns was used for the isolation of the phytoestrogens from the matrix. An extraction on a ChemElut 1010 column connected on-line to a Florisil cartridge by a Teflon stopcock was used for effective eluate purification. A mixture of dichloromethane and ethyl acetate was used for elution of the isoflavones from the columns in tandem. The isoflavones were determined as trimethylsilyl (TMS) ethers using GC/MS-SIM after separation on an HP-5MS fused silica column. TMS ethers were obtained by using BSTFA containing 1% of TMCS. For the determination of free isoflavones 6-hydroxyflavone was used as internal standard, whereas robigenin was used in the case of total isoflavone determination. Recoveries for free isoflavones under study varied from 63.5 to 89.6% at the 25 ng mL(-)(1) level and from 63.5 to 89. 2% at the 5 ng mL(-)(1) level in urine. Analytical curves were linear between 5 and 25 ng mL(-)(1). Detection limits varied from 1 ng mL(-)(1) for formononetin to 2.3 ng mL(-)(1) for daidzein. Recoveries for total isoflavone determination after enzymatic hydrolysis with glucuronidase from Helix pomatia ranged from 56.5 to 77.1% at the 25 ng mL(-1) level.  相似文献   

12.
Doramectin (DRM) is a broad spectrum macrocyclic lactone antiparasitic drug not approved for use in dairy animals. However, DRM and other endectocide compounds are widely used extra-label to control endo- and ectoparasites in dairy sheep. The plasma disposition kinetics and the pattern of DRM excretion in milk were characterized following its subcutaneous administration to lactating dairy sheep. DRM concentration profiles were measured in plasma and milk samples after validation of a specific HPLC-based methodology. DRM was detected between 1 h and 30 days post-treatment. DRM concentrations of 0.48 ng.mL(-1) (plasma) and 1.03 ng.mL(-1) (milk) were measured at 30 days post-treatment. DRM was extensively distributed from the bloodstream to the mammary gland, and large concentrations were excreted in milk. The peak concentrations and total amount of DRM recovered in milk (expressed as area under the concentration versus time curve) were 3-fold higher than those measured in plasma; 2.44% of the total DRM dose was excreted in milk. The long persistence of DRM milk residues should be seriously considered before its extra-label use in dairy animals is recommended.  相似文献   

13.
A competitive indirect enzyme-linked immunosorbent assay (ciELISA) for the organophosphorus insecticide acephate, O,S-dimethyl acetylphosphoramidothioate, was developed using a polyclonal antibody. Five different haptens mimicking the analyte were synthesized and conjugated with the carrier proteins bovine serum albumin (BSA) and keyhole limpet hemocyanin (KLH) by the N-hydroxysuccinimide active ester and diazotization methods. Polyclonal antibodies raised against hapten-KLH conjugates in rabbits and hapten-BSA conjugates as coating antigens were screened and selected for the assay in the homologous and heterologous ELISA systems. The effects of various assay conditions such as detergent, organic solvents, pH, and preincubation of the mixture of the polyclonal antibody and the analyte on the sensitivity were evaluated. The IC(50) value for acephate was 25 ng/mL in an optimized heterologous system using hapten-4-BSA as a coating antigen and a polyclonal antibody no. 8377 against hapten-1-KLH, showing the detection range of 5-140 ng/mL and the lowest detection limit of 2 ng/mL. The cross-reactivities of the structurally related organophosphorus insecticides, including the major metabolite of the analyte, methamidophos, were less than 1%. Recoveries from the analyte-fortified tap water, mulberry leaves, and lettuce samples in the assay were in the range of 72-121% by simple extraction, concentration, and dilution. These results indicate that the ELISA could be a convenient and supplemental analytical tool for monitoring acephate residues in environmental and agricultural samples.  相似文献   

14.
Six new compounds, trans-3-isobutyl-4-[4-(3-methyl-2-butenyloxy)phenyl]pyrrolidine-2,5-dione (1), trans-1-hydroxy-3-(4-hydroxyphenyl)-4-isobutylpyrrolidine-2,5-dione (2), cis-3-(4-hydroxyphenyl)-4-isobutyldihydrofuran-2,5-dione (3), 3-(4-hydroxyphenyl)-4-isobutyl-1H-pyrrole-2,5-dione (4), 3-(4-hydroxyphenyl)-4-isobutylfuran-2,5-dione (5), and dimethyl 2-(4-hydroxyphenyl)-3-isobutylmaleate (6), together with one known compound, 3-isobutyl-4-[4-(3-methyl-2-butenyloxy)phenyl]furan-2,5-dione (7), were isolated from the fruiting bodies of Antrodia camphorata. The structures of the compounds were elucidated by analysis of their spectroscopic data. To investigate the immunomodulatory potential of the compounds, RAW264.7 macrophage cells were treated with the compounds. Compound 1 significantly increased spontaneous TNF-alpha secretion from unstimulated RAW264.7 cells but suppressed IL-6 production [50% inhibition concentration value (IC50) = 10 microg/mL] in LPS-stimulated cells. Compounds 3, 4, and 6 also suppressed IL-6 production with IC50 values of 17, 18, and 25 microg/mL, respectively, suggesting that these four compounds may have an anti-inflammatory effect on macrophage-mediated responses. Of the six compounds, compound 1 was the most effective, exerting both immunostimulatory and anti-inflammatory effects.  相似文献   

15.
硼砂对绵羊氮代谢及营养物质消化的影响   总被引:1,自引:3,他引:1  
刘大森  单安山  张鹏 《核农学报》2006,20(5):438-440
本文研究了硼砂对绵羊营养物质消化率及氮代谢的影响。选择绵羊4只,采用4×4拉丁方设计,预饲期10d,试验期7d。在处理组的日粮中分别添加1g硼砂/kg、2g硼砂/kg和25mg乙酰氧肟酸/kg,结果表明,添加硼砂对日粮干物质、有机物和粗蛋白质消化率及对氮和尿素代谢均未产生影响。1g/kg日粮的硼砂添加量促进了瘤胃微生物氮合成。试验结果提示绵羊对脲酶抑制剂硼砂也存在适应性。  相似文献   

16.
A simple, sensitive, and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determining urinary creatinine was developed and used to evaluate 24 h urine samples collected during an exposure study. Urine (1 microL) was diluted with methanol and then directly applied to LC-MS/MS. Under electrospray ionization (ESI) conditions, the transition molecules of creatinine and creatinine- d3 were observed at m/ z 114 > 44 and m/ z 117 > 47, respectively. The retention time of creatinine was 0.59 min. The linear range was 1-2000 ng/mL, with a detection limit in urine of 1 ng/mL. LC-MS/MS and colorimetric end-point methods were significantly associated ( R2 = 0.8785, p < 0.0001). The LC-MS/MS method to determine creatinine in 24 h urine samples had shorter retention times, was more sensitive, reliable, reproducible, simple, selective, and used a smaller sample size than other LC-MS/MS or commercial methods.  相似文献   

17.
The distribution of ivermectin in buffalo plasma and milk after administration of a single subcutaneous dose (0.2 mg kg(-)(1) b.w.) was studied. Ivermectin reached the maximal concentration in plasma (28.5 +/- 1.7 ng mL(-)(1)) and milk (23.6 +/- 2.6 ng mL(-)(1)) after 2.4 +/- 0.32 and 2.8 +/- 0.44 days, respectively. The drug showed a parallel disposition in milk and plasma, with a ratio of 1.12 +/- 0.16. Ivermectin concentrations were detected in mozzarella cheese obtained from milk collected on days 1, 3, 4, and 20 following administration. The highest values (81.4 +/- 3.26 ng g(-)(1)) were found in the cheese produced on day 3 and were 4-fold higher than those present in the milk.  相似文献   

18.
Previous research has demonstrated transmission of zearalenone and alpha- and beta-zearalenols into the milk of cows and other animals. Since human intake of zearalenone and its metabolites via milk is an unknown factor in risk assessment of zearalenone and because appropriate methodology for their determination in milk is not available, a rapid and sensitive analytical method has been developed. Essentially, the method includes extraction with basic acetonitrile, acidification, partition into methylene chloride on a hydrophilic matrix, cleanup on an aminopropyl solid phase extraction column, and reverse-phase liquid chromatography with fluorescence detection. Recoveries from milk averaged 84% for zearalenone, 93% for alpha-zearalenol, and 90% for beta-zearalenol at spiking levels of 0.5 to 20 ng/mL. As little as 0.2 ng/mL of zearalenone and alpha-zearalenol and 2 ng/mL of beta-zearalenol can be detected in milk. These 3 compounds are stable in refrigerated milk for at least 2 weeks and in milk brought to boiling. Enzymes (beta-glucuronidase and aryl sulfatase) may be added to milk prior to extraction to hydrolyze any conjugates.  相似文献   

19.
A biosensor method, using the surface plasmon resonance (SPR) principle, was developed for the determination of ractopamine in cattle and sheep urine. A monoclonal antibody was used to compete with ractopamine in the sample and ractopamine immobilized on the sensor chip. Addition of bovine serum albumin (BSA, 1 mg/mL) as an antibody stabilizer to the incubation buffer was required to achieve a stable biosensor response throughout each sample set. The calibration curve gave a mean IC(50) of 4.7 +/- 0.21 ng/mL (n = 7). Over sample concentrations from 2.5 to 10 ng/mL recoveries were typically approximately 100-110%, whereas inter- and intra-assay reproducibilities (% CV) were usually less than 10 and 6%, respectively. Comparison of biosensor results with results obtained from high-performance liquid chromatography (HPLC) and enzyme-linked immunosorbent assays (ELISA) using enzyme-hydrolyzed urine (to convert ractopamine conjugates to free ractopamine) gave correlation coefficients of 0.94 for sheep and 0.86 for cattle. Slopes of the lines, with zero intercepts, equaled 0.80 for sheep and 0.74 for cattle. For untreated (nonhydrolyzed) urine samples, the correlations between biosensor and HPLC results were 0.95 for sheep and 0.72 for cattle with slopes of 1.18 (sheep) and 1.69 (cattle). The slopes greater than unity indicate that the biosensor responded to ractopamine metabolites in addition to free ractopamine. The biosensor assay is an excellent analytical tool to screen ractopamine residues in sheep or cattle urine, and the results should be extendible to other species with suitable validation.  相似文献   

20.
trans-Caftaric acid is the most abundant nonflavonoid phenolic compound in grapes and wines. It occurs in chicory and is one of the bioactive components of Echinacea purpurea. In order to fill the gap of knowledge about its bioavailability in mammals, we investigated its absorption, tissue distribution, and metabolism in rats. Assuming that the stomach is a relevant site of absorption of dietary polyphenols, a solution of trans-caftaric acid was maintained in the ligated stomach of anaesthetized rats for 20 min. Intact trans-caftaric acid was detected in rat plasma at both 10 and 20 min (293 +/- 45 and 334 +/- 49 ng/mL, respectively), along with its O-methylated derivative trans-fertaric acid, whose concentration rose over time (from 92 +/- 12 to 185 +/- 24 ng/mL). At 20 min, both trans-caftaric acid and trans-fertaric acid were detected in the kidney (443 +/- 78 and 2506 +/- 514 ng/g, respectively) but not in the liver. Only trans-fertaric acid was found in the urine (33.3 +/- 12.8 microg/mL). In some rats, trans-caftaric acid was detected in the brain (180 +/- 20 ng/g).  相似文献   

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