Category Archives: elisa

Analysis of Total Ergot Alkaloid Amount in Various Grain Samples by ELISA

By Dr. Frederik Wuppermann and Dr. Uwe Aulwurm

Introduction:

Ergot alkaloids are mycotoxins produced by fungi (Claviceps sp.) on various grain species with a highly toxic impact on the health of consumers (Figure 1). The toxins provoke symptoms summarized as ergotism.

ergot wheat

Figure 1: Sclerotia grown on crop

The toxins are characterized by two different isomers, which occur under different pH conditions, they are named –ines and –inines. The main toxins found are ergocristine, -inine, ergometrine, -inine, ergosine, -inine, ergocryptine, -inine, ergotamine, -inine, ergocornine, -inine. The assessment of toxin amount becomes necessary due to the toxic effects of ergot alkaloids. For this purpose an ELISA was developed to detect all ergot alkaloids in various crop material and the detected amounts of ergot alkaloids were correlated with LC-MS data.

 

 

Method

ergoread quick method
Figure 2.
ELISA procedure

The sample is extracted by 40/60 methanol / 0.25% phosphoric acid as described by Ware et al. 2000. After centrifugation the sample was diluted with sample dilution buffer and filtered again. The filtered sample was applied in the ELISA plate and compared with a standard solution (enclosed in the ErgoREAD kit). The enzyme immune reaction is performed as depicted in Figure 2.

 

 

Results:

Analysis of wheat samples:

analysis of wheat

 Analysis of Rye Samples:

analysis of rye

Comparative Analysis of Naturally Contaminated Samples:

The samples showed good correlation between 0 to 10 ppm, indicating that the ELISA is suitable for the screening of various crop material for the presence of ergot alkaloids. The partially over quantification in the comparative analysis could be due to the cross reactivity of the ELISA against various ergot alkaloids which were not monitored by the LC-MS or HPLC analysis. comparative analysis of naturally contaminated samples

Figure 3 cross reactivity of ErgoREADThe ELISA shows significant cross reactivity against various ergot alkaloids e.g. ergonovin (ergometrine or ergocristine) and can be used to determine the qualitative stuatus of a crop sample concernig the sum of ergot alkaloids.

 Conclusion:

The ErgoREAD ELISA is an appropriate tool for screening wheat, rye and triticale samples for presence of ergot alkaloids. The range of measurement is from 0 to 5 ppm. Due to the readout as the sum of all reactive ergot alkaloids, the cutoff for critical samples is 0.4 ppm. All shown ELISA samples are correlated with HPLC. The total amount of samples which need to undergo LC-MS analysis could be dramatically reduced.

More information about this kit and other Mycotoxin analysis products can be found on LCTech’s Website. The original poster can be downloaded here.

Pickering Laboratories is proud to bet the distributor for LCTech in North America. For more information about our long-time relationship with LCTech, check out our blog entry on Pickering’s Company & Product Offerings in 2013.

 

 

Pickering Laboratories in 2013

By Wendy Rasmussen

In recent years, we have released several new products and applications, and with still more on the horizon, it occurred to me that now would be a great time to summarize the Pickering of today – our mindset and our wide variety of products & applications.

Acai Berries
Acai Berries

No longer are we simply the “Post-Column Company”. We are the “Automated-Sample Antioxidants” company.  Think of us as the new “super fruit.” The Acai berry, or perhaps the new Chia Seeds (incidentally, we do have a post-column application for the identification & quantitation of  Antioxidants in a variety of matrices).

We are still very active, and we as a company plan to be here for many years to come. We are still the company founded on chemistry and a desire to to teach, to spread our technical expertise, to support our customers.

It’s been a few years now since Pickering began distributing and supporting our LCTech Product line.  The products have shown an ever increasing interest here in the US and in Canada (our official Sales Territory for this product line). We are very proud to offer these products and we hope we can develop it further in the future.

Historically, we have provided the back-end of an analysis (post-column derivatization). Nowadays, we can provide the front-end of analysis as well (the sample-cleanup).

In thinking about our product offerings, I realized that a simple list does not effectively show the scope of the products we have to offer in 2013 – primarily because we have a lot of overlap between products and product lines.  We’re not a vertical company in that regard.  I suppose one could say our product offerings are more circular in that many do not fall into a single distinct category.  I am a very visual person, and for me, a Venn diagram and our overall “product scale” really helped to understand and clarify our products:

Venn Diagram of Pickering Laboratories
Venn Diagram of Pickering Laboratories
General "Product Scale" for Pickering Offerings
General “Product Scale” for Pickering Offerings
For those of who like lists, you can find one Here, on Pickering’s website, and on LCTech’s Website

For any Questions, please feel free to contact us:

Pickering Laboratories, Inc.
Mountain View, California
Email: sales@pickeringlabs.com
Phone: (direct) 650-694-6700 or (toll-free) 800-654-3330

 

Image of Acai Berries:  http://acaiberryeducation.com/

Pickering Products

Mycotoxins: Clean-up Columns, ELISA Kits, Post-Column Derivatization Instruments & Methods, SPE manifolds (manual & automated) for the analysis of:

Aflatoxins
Ochratoxin A
DON
Nivalenol
Fumonisin FB1, FB2
Zearalenone
Ergot Alkaloids

Post-Column Derivatization: Pinnacle PCX, Vector PCX, UVE Photochemical Reactor, Columns, Reagents, Eluants for the analysis of:

Antioxidants
Mycotoxins (individual and multi-residue)
Amino Acids
Biogenic Amines
Paralytic Shellfish Toxin
Polyether Antibiotics
Hexavalent Chromium
Vitamins
Sugars
Nitrate/Nitrite
Bromate
Carbamate Pesticides
Glyphosate Herbicide
NDELA
Formaldehyde

Sample Preparation (clean-up) Instruments:

FREESTYLE for SPE, GPC, online-concentration
FREESTYLE ThermELUTE for Aflatoxin analysis (direct inject onto HPLC)
AcceCLEAN for SPE (incl. Immunoaffinity columns)
EluVAC vacuum manifold for SPE (incl. Immunoaffinity columns)
GPC QUATTRO for manual GPC Cleanup
DECS System for the cleanup of samples for Dioxin analysis

Chemistry Products:

Artificial Perspiration
Artificial Saliva
High Purity Water

Stand-Alone OEM Syringe Pump:

For any applications requiring an inert flow path, volumes up to 70mL, and pressures up to 500psi

For More Information visit:

www.pickeringlabs.com

or email: sales@pickeringlabs.com