scholarly journals Covariation of Ergot Severity and Alkaloid Content Measured by HPLC and One ELISA Method in Inoculated Winter Rye across Three Isolates and Three European Countries

Toxins ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 676
Author(s):  
Anna Kodisch ◽  
Michael Oberforster ◽  
Armin Raditschnig ◽  
Bernd Rodemann ◽  
Anna Tratwal ◽  
...  

Ergot caused by Claviceps purpurea is a problem for food and feed security in rye due to the occurrence of toxic ergot alkaloids (EAs). For grain elevators and breeders, a quick, easy-to-handle, and cheap screening assay would have a high economic impact. The study was performed to reveal (1) the covariation of ergot severity (= percentage of sclerotia in harvested grain) and the content of 12 EAs determined by high performance liquid chromatography (HPLC) and (2) the covariation between these traits and results of one commercial enzyme linked immunosorbent assays (ELISA). In total, 372 winter rye samples consisting of a diverse set of genotypes, locations from Germany, Austria, and Poland over two years, and three isolates were analyzed. Ergocornine and α-ergocryptine were detected as major EAs. Ergocristinine occurred as a minor component. Claviceps isolates from different countries showed a similar EA spectrum, but different quantities of individual EAs. A moderate, positive covariation between ergot severity and EA content determined by HPLC was observed across two years (r = 0.53, p < 0.01), but large deviation from the regression was detected. ELISA values did neither correlate with the HPLC results nor with ergot severity. In conclusion, a reliable prediction of the EA content based on ergot severity is, at present, not possible.

1980 ◽  
Vol 26 (6) ◽  
pp. 707-711 ◽  
Author(s):  
T D Schlabach ◽  
J A Fulton ◽  
P B Mockridge ◽  
E C Toren

Abstract We observed nonenzymic peaks when serum isoenzymes of lactate dehydrogenase (EC 1.1.1.27; LD) and creatine kinase (EC 2.7.3.2.; CK) were separated by “high-performance” liquid chromatography and detected by continuously monitoring the column effluent for enzyme activity. Such background peaks were particularly apparent in CK isoenzyme profiles obtained from human sera. We observed two nonenzymic peaks with fluorescence detection, one in the CK-MB region, the other in the CK-BB region. Serum albumin was a major component in the artifactual CK-MB peak, with lipoprotein as a minor component. We present evidence that the material responsible for the other peak fluoresced quite strongly and is mostly pre-albumin.


2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Reem H. Obaydo ◽  
Duaa J. Al Zakri ◽  
Amir Alhaj Sakur ◽  
Hayam M. Lotfy

Abstract Background The pharmaceutical industry and the National Regulatory Authorities are now focusing on the quantification of multi-component drugs for quality control testing. Main body Recently, the utilization of the ultraviolet spectrophotometric methods has become vital for the analytical studies for the routine analysis of different fixed-dose combination products either in the presence of a minor component in their combination or not. This article reviewed several published methods to those that have been applied to quantify some pharmaceutical combinations containing minor components through the last two decades. Conclusion The applied spectrophotometric methods are labor, time-saving, and also considered a cheap substitute for the overpriced high-performance liquid chromatographic technique.


1986 ◽  
Vol 240 (3) ◽  
pp. 625-632 ◽  
Author(s):  
K G Jacobsson ◽  
U Lindahl ◽  
A A Horner

Rat skin heparin proteoglycan labelled biosynthetically with 35S was fractionated on a column of antithrombin-Sepharose into fractions with varying degrees of affinity for antithrombin. These were treated with NaOH to release heparin chains (Mr 60,000-100,000), by beta-elimination or incubated with serum to produce fragments of the same order of size as commercial heparin (Mr 5000-30,000), by endoglycosidase cleavage. Chains and fragments were then fractionated on antithrombin-Sepharose. The various fractions were deaminated with HNO2 at pH 1.5 followed by reduction with NaB3H4. Approx 90% of the incorporated 3H was associated with disaccharides. These were fractionated by high-performance ion-exchange chromatography. A unique minor component corresponding to the sequence glucuronosyl-N-sulphoglucosaminyl (3,6-di-O-sulphate) in the polysaccharide was found only in fractions with high affinity for antithrombin. The glucosamine residue linked to C-4 of this glucuronosyl unit was predominantly (or exclusively) N-sulphated rather than N-acetylated, pointing to a structural difference between the antithrombin-binding region of rat heparin and that of pig mucosal heparin. Calculations based on the distribution of the glucosaminyl 3-O-sulphate group showed that approximately two-thirds of the total antithrombin-binding regions present in the unfractionated material were accommodated by only 20% of the proteoglycan molecules, and by 10% of the polysaccharide chains. While most of the proteoglycan molecules thus lacked such regions (and hence affinity for antithrombin) a minor proportion of the polysaccharide chains contained on the average three binding regions per molecule. These findings support by direct chemical analysis an earlier proposal, based on anticoagulant activities of similar rat skin heparin fractions, that the distribution of antithrombin-binding sites in intact heparin proteoglycans is markedly non-random.


1980 ◽  
Vol 26 (6) ◽  
pp. 707-711
Author(s):  
T D Schlabach ◽  
J A Fulton ◽  
P B Mockridge ◽  
E C Toren

Abstract We observed nonenzymic peaks when serum isoenzymes of lactate dehydrogenase (EC 1.1.1.27; LD) and creatine kinase (EC 2.7.3.2.; CK) were separated by “high-performance” liquid chromatography and detected by continuously monitoring the column effluent for enzyme activity. Such background peaks were particularly apparent in CK isoenzyme profiles obtained from human sera. We observed two nonenzymic peaks with fluorescence detection, one in the CK-MB region, the other in the CK-BB region. Serum albumin was a major component in the artifactual CK-MB peak, with lipoprotein as a minor component. We present evidence that the material responsible for the other peak fluoresced quite strongly and is mostly pre-albumin.


2009 ◽  
Vol 74 (10) ◽  
pp. 1543-1557 ◽  
Author(s):  
Herman P. Van Leeuwen ◽  
Raewyn M. Town

The degree of (de)protonation of aqueous metal species has significant consequences for the kinetics of complex formation/dissociation. All protonated forms of both the ligand and the hydrated central metal ion contribute to the rate of complex formation to an extent weighted by the pertaining outer-sphere stabilities. Likewise, the lifetime of the uncomplexed metal is determined by all the various protonated ligand species. Therefore, the interfacial reaction layer thickness, μ, and the ensuing kinetic flux, Jkin, are more involved than in the conventional case. All inner-sphere complexes contribute to the overall rate of dissociation, as weighted by their respective rate constants for dissociation, kd. The presence of inner-sphere deprotonated H2O, or of outer-sphere protonated ligand, generally has a great impact on kd of the inner-sphere complex. Consequently, the overall flux can be dominated by a species that is a minor component of the bulk speciation. The concepts are shown to provide a good description of experimental stripping chronopotentiometric data for several protonated metal–ligand systems.


2021 ◽  
Vol 1 (1) ◽  
pp. 11-23
Author(s):  
Arjun Bhusal ◽  
Peter M. Muriana

In the US, sodium nitrate is used as a preservative and curing agent in processed meats and is therefore a regulated ingredient. Nitrate reducing bacteria (NRB) can convert vegetable nitrate into nitrite allowing green/clean label status in the US as per the USDA-FSIS definition of ‘natural nitrite’. The current ‘in-liquid’ test tube assay for detecting nitrite is not suitable for screening mixtures of bacteria nor is commercial nitrate broth suitable for growth of many Gram (+) bacteria. M17 broth was therefore used to develop M17-nitrate broth to be inclusive of Gram (+) bacteria. An ‘on-agar’ colony-screening assay was developed to detect the conversion of nitrate to nitrite on agar plates and could detect one NRB+ colony among ~300–500 colonies on a single plate. Samples that might have NRB were spread-plated on M17 agar plates, sandwiched with nitrate agar, and after incubation followed with sequential agar overlays containing the reagents used in the nitrate reduction assay; the appearance of red color zones above colonies indicated the presence of nitrite. NRB derived from various samples were confirmed for nitrate conversion and both nitrate and nitrite were quantified by C8 reversed-phase (RP) ion-pairing high performance liquid chromatography (HPLC) analysis (1 ppm limit of detection). Staphylococcus carnosus, a strain commonly used for nitrate reduction, was able to convert 1100 ppm M17-nitrate broth to 917 ppm nitrite. Staphylococcus caprae and Panteoa agglomerans, NRB isolated using the M17-nitrate agar assay, were also able to ferment the same broth to 916 ppm and 867 ppm nitrite, respectively. This is the first report of an on-agar colony screening assay for the detection and isolation of nitrite reducing bacteria allowing NRB to be readily isolated. This may allow for the identification of new bacteria that may have a more efficient process to generate nitrite, and possibly concomitant with production of additional natural antimicrobials, as vegetable nitrite becomes more widely used to prevent spore germination.


1994 ◽  
Vol 6 (4) ◽  
pp. 517-527 ◽  
Author(s):  
Duncan Pirrie

Late Cretaceous sedimentary rocks assigned to the Santa Marta (Herbert Sound Member) and López de Bertodano (Cape Lamb and Sandwich Bluff members) formations of the Marambio Group, crop out on Cape Lamb, Vega Island. Although previous studies have recognized that these sedimentary rocks were derived from the northern Antarctic Peninsula region, the work presented here allows the provenance and palaeogeographical evolution of the region to be described in detail. On the basis of both sandstone petrography and clay mineralogy, the Herbert Sound and Cape Lamb members reflect sediment input from a low relief source area, with sand grade sediment sourced from low grade metasediments, and clay grade sediment ultimately derived from the weathering of an andesitic source area. In contrast, the Sandwich Bluff Member reflects a switch to a predominantly andesitic volcaniclastic source. However, this sediment was largely derived from older volcanic suites due to renewed source area uplift, with only a minor component from coeval volcanism. Regional uplift of both the arc terrane and the western margin of the James Ross Basin was likely during the Maastrichtian.


1988 ◽  
Vol 254 (1) ◽  
pp. 67-71 ◽  
Author(s):  
B Rüstow ◽  
Y Nakagawa ◽  
H Rabe ◽  
K Waku ◽  
D Kunze

1. Phosphatidylinositol (PI) is a minor component of lung surfactant which may be able to replace the functionally important phosphatidylglycerol (PG) [Beppu, Clements & Goerke (1983) J. Appl. Physiol. 55, 496-502] without disturbing lung function. The dipalmitoyl species is one of the main species for both PI (14.4%) and PG (16.9%). Besides the C16:0--C16:0 species, the C16:0--C18:0, C16:0--C18:1, C16:0--C18:2 and C18:0--C18:1 species showed comparable proportions in the PG and PI fractions. These similarities of the species patterns and the acidic character of both phospholipids could explain why surfactant PG may be replaced by PI. 2. PI and PG were radiolabelled by incubation of microsomal fractions with [14C]glycerol 3-phosphate (Gro3P). For 11 out of 14 molecular species of PI and PG we measured comparable proportions of radioactivity. The radioactivity of these 11 species accounted together for more than 80% of the total. The addition of inositol to the incubation system decreased the incorporation in vitro of Gro3P into PG and CDP-DG (diacylglycerol) of lung microsomes (microsomal fractions), but did not change the distribution of radioactivity among the molecular species of PG. These results supported the idea that both acidic surfactant phospholipids may be synthesized de novo from a common CDP-DG pool in lung microsomes.


Sign in / Sign up

Export Citation Format

Share Document