Category Archives: HPLC

Amino Acid Analysis, HPLC, pickering laboratories, pinnacle pcx, post-column derivatization, Trione

Amino Acid Analysis of Monoclonal Antibodies

Method Abstract 373, Amino Acid Analysis of Monoclonal Antibodies

The peptide and protein based pharmaceuticals are a rapidly expanding class of therapeutical agents that are used to treat a wide variety of health conditions, including cancer, metabolic and auto-immune diseases, HIV and more. Biologic drugs, such as monoclonal antibodies, are derived from living organisms and are usually very expensive. As many biologics are coming off of patents, the market is ready for cost-saving biogenerics. But all proteins, including monoclonal antibodies, have complex structures that determine their function. Differences in structure would alter biological activity leading to changes in safety and efficacy of the drug.

ICH Q6B is a guidance document that provides a set of internationally accepted specifications for biotechnological and biological products to support new marketing applications. It establishes the set of criteria to which a drug substance, drug product or material should conform to be considered acceptable for intended use.

aaa chromatograms from MA373Determining Amino Acid composition following hydrolysis is listed in ICH Q6B as a way to characterize the protein and to confirm its identity by comparing with Amino Acid composition deduced from the gene sequence of the desired product. Amino Acid Analysis data is also used to accurately determine the protein content.

The Amino Acids Analysis with post-column derivatization is a very sensitive, reproducible and rugged method and it has been a preferred approach for laboratories running biological samples, protein, peptides and foods analysis. Pickering Laboratories Inc. offers many Amino Acids Analysis products including post-column derivatization instruments, columns, eluants, reagents and standards. All products are designed to work together to deliver optimum results for any chosen sample.

METHOD
Analytical conditions
Column: High-efficiency Sodium cation-exchange column, 4.6 x 110 mm, P/N 1154110T
Flow Rate: 0.6 mL/min
Mobile Phase: See method in Table 1

Post-Column Conditionsgradient table for AAA monoclonal AB
Post-column System: Pinnacle PCX
Reactor Volume: 0.5 mL
Reactor Temperature: 130 °C
Flow Rate: 0.3 mL/min
Detection: UV/VIS 570 nm for primary amino acids, 440 nm for secondary amino acids
Injection Volume: 10-50 uL

You can download this abstract, as well as our product catalog and other notes from our website: www.pickeringlabs.com

 

aflatoxin, Amino Acid Analysis, AOAC, clean-up, deoxynivalenol, freestyle, HPLC, immunoaffinity cleanup, mycotoxin, ochratoxin

AOAC International Meeting

fountainThe AOAC International held their annual meeting in Chicago, Illinois from August 25-27, 2013.

Pickering Laboratories were on hand to share our latest methods and instruments.  We also gave an Exhibitor Presentation titled “An Automated Highly Sensitive Method for Aflatoxin B/G or Ochratoxin A Clean-up and Analysis – A Novel Approach in Mycotoxin Analysis”

The purpose of the talk was to showcase our newest method and instrument:

Building on the FREESTYLE Automated Sample Preparation Instrument, the new ThermELUTE module and columns enable rapid, sensitive analysis of Aflatoxin or Ochratoxin A. The ThermELUTE module uses a thermal denaturation technique to release jellybean4toxin from a new column format requiring only 20% of usual solvent volumes, and FREESTYLE quantitatively transfers the eluate into the HPLC. In combination with online injection, analytical sensitivity can be increased without manual manipulation. The increase in sensitivity provides a reduction of sample load, while maintaining excellent chromatographic results. The increase of sensitivity allow analysis of mycotoxin levels well below regulatory limits.

The ThermELUTE takes the separate steps of cleanup and analysis, and combines them into one automated system. Using a FREESTYLE robotic system equipped with the

FREESTYLE Automated Sample Preparation System
FREESTYLE Automated Sample Preparation System

ThermELUTE module, we can now clean up a sample for mycotoxins, and then directly inject the cleaned sample onto the HPLC column for analysis!

Automating Mycotoxin Analyses from Cleanup to Detection:

  • Results 3 times faster
  • 80% less solvent
  • Direct connect to HPLC

The brochure for ThermELUTE can be found HERE. To view the slides from our presentation, click HERE.

 

AOAC, HPLC, pickering laboratories, pinnacle pcx, post-column derivatization

Polyether Antibiotics in Animal Feed

Our updated Method Abstract 264 describes the analysis of Monensin, Salinomycin, Narasin, & Lasalocid in Animal Feeds.

Polyether Antibiotics are commonly used for preventing coccidiosis and other infections in poultry and for improving feed efficiency for beef cattle and swine. The use of Polyether Antibiotics is strictly regulated, with only specific ionophores approved for use in feeds intended for different animals.

Analysis of Polyether Antibiotics by HPLC with post-column derivatization and UV/Vis detection has been proven to successfully identify and quantify Monensin, Narasin and Salinomycin in medicated feeds, supplements and premixes as well as to determine trace contamination levels in non-medicated feeds [1, 2].

Post-column derivatization of Polyether Antibiotics is done using highly acidic Vanillin or DMAB reagents. The Pinnacle PCX derivatization system (Pickering Laboratories, Inc.) has an inert flow path and automated system wash capabilities that make it uniquely suitable for handling corrosive reagents. The two-pump system is recommended to extend reagent stability, but the single-pump system for this application is also available.

Adding a Fluorescence detector to the instrumentation allows for using the same extraction procedure and HPLC conditions to also determine Lasalocid, which doesn’t require post-column derivatization.

Chromatograms of Polyether AntibioticsMETHOD
Sample Preparation
To 25 g of finely ground feed sample, add 100 mL of extraction solution (90% Methanol – 10% water). Shake for 1 hour at high speed using a mechanical shaker. Let the solids settle and filter an aliquot of the extract for injection. Dilute with extraction solution if needed to fit the calibration curve. Use a 2.5 g portion when testing premixes.

Analytical conditions
Analytical Column: Polyether Column, C18, 4.6 x 250 mm, Catalog No 2381750
Temperature: 40 ºC
Flow Rate: 0.7 mL/min
Mobile Phase: 90% Methanol, 10% of 5% Acetic Acid solution in water, isocratic
Injection volume: 20 μL

Post-Column Conditions
Post-column System: Pinnacle PCX
Reactor Volume: 1.4 mL
Reactor Temperature: 90 °C
Reagent 1: Concentrated Sulfuric acid / Methanol (4:96 v/v)
Reagent 2: 60 g of Vanillin in 950 mL of Methanol
Reagent Flow Rate: 0.3 mL/min
Detection: UV/VIS 570 nm (for Lasalocid – FLD, Ex. 322 nm, Em. 370 nm)

Conclusion
Analysis of Polyether Antibiotics by HPLC with Post-column derivatization is a robust and sensitive method that utilizes standard equipment and could easily be adopted by testing laboratories. It allows for testing of different ionophores at wide range of concentrations,
including at trace levels. Using Pinnacle PCX post-column derivatization system, factory configured for the analysis, guarantees stable and reproducible results.

The full application note as well as our complete Product Catalog can be downloaded from our website: www.pickeringlabs.com

table for polyether antibiotics

AcceCLEAN, aflatoxin, amino acid, Amino Acid Analysis, amino acids, antioxidants, AOAC, AOAC's Laboratory of the Year Award, artificial perspiration, carbamate, Chromatography, clean-up, column, deoxynivalenol, detector, dietary supplements, DON, Dr. Pickering, elisa, eluvac, ergot alkaloid, freestyle, GARD, glyphosate, GPC, Guard, HPLC, immunoaffinity cleanup, Multi-residue Mycotoxin Analysis, mycotoxin, natural toxins, New Products, Ninhydrin, ochratoxin, paralytic shellfish toxin, Physiological Fluids, pickering laboratories, pinnacle, pinnacle pcx, Post-column, post-column derivatization, psp, sweat, syringe pump, Trione, UVE, vector

Pickering Laboratories in 2013

1 Comment

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/

aflatoxin, Amino Acid Analysis, carbamate, chromatographic specialties, Chromatography, clean-up, eluvac, freestyle, glyphosate, GPC, herbal products, HPLC, immunoaffinity cleanup, lctech, Multi-residue Mycotoxin Analysis, mycotoxin, natural products, natural toxins, New Products, Ninhydrin, ochratoxin, pickering laboratories, pinnacle pcx, Pittcon, post-column derivatization, spe

Pickering Exhibits at Pittcon 2013

This year’s Pittcon was held in Philadelphia, Pennsylvania from March 17-21, 2013.

view of philly from the museum On display were Pinnacle PCX, EluVAC, and a video of the FREESTYLE Automated Sample Preparation Instrument.  The video was a new technique for us, and it was well received.

The video in our booth was a combination of the separate videos for each module of the FREESTYLE Automated Sample Preparation Instrument. You can view the originals here (they are very informative, and we highly recommend them): FREESTYLE VIDEOS

Wendy, Mike and David enjoyed visiting with existing and future customers, as well as our distributors from around the world, including our Canadian distributor, Chromatographic Specialties.

Before the show opened, we had the opportunity to explore the city.  Philadelphia is one of our oldest cities and is full of history and landmarks. Some of this editor’s favorites include Independence Hall and the surrounding neighborhoods, and the Art Museum (see more below).  On this trip, we explored Reading Terminal Market, and sampled that famous cuisine, the Philly Cheese steak Sandwich. I’m told it is not truly a Philly Steak sandwich unless you have it with Cheez Whiz. Any type of real cheese doesn’t count. I made that mistake on my first trip to Philadelphia, and the man behind the counter let me know it!  I wasn’t about to make that mistake again:

philly cheesesteak

The Philadelphia Museum of Art is known for its amazing collection of artwork (viewed on a previous visit), but is also made famous by the scene of Sylvester Stallone running up the stairs in the movie Rocky.  Since the author of this entry happened to be training for a race, she decided that a little cold wasn’t going to stop her. So she put on her iPod (Rocky Theme playing, of course), and ran from the hotel to the top of the steps and back (total distance: about 2.8miles).

view from the bottom

wendy philly top of steps

wendy rockyOn the return to the hotel, I came across a version of the LOVE sculpture, by American Artist Robert Indiana. I always thought it was was much bigger…

love sculpture

 

For those of you interested in learning more about the sculpture, click HERE

If you’re ever in Philadelphia, the Philadelphia Museum of Art is definitely worth a visit.

But if you can’t make the trip to Philadelphia, you could also stay home and watch Rocky…

aflatoxin, deoxynivalenol, DON, HPLC, immunoaffinity cleanup, Multi-residue Mycotoxin Analysis, mycotoxin, ochratoxin, pinnacle pcx, post-column derivatization, UVE, vomitoxin

On-Demand Webinar: Rugged and Sensitive Analysis of Mycotoxins in Food Matrices

If you missed the live webinar last summer, you can still hear the Webinar On-Demand! Just click on the links below to launch the presentation.
On August 14th, 2012, in collaboration with Thermo Scientific, Pickering Laboratories participated in a free webinar titled: Rugged and Sensitive Analysis of Mycotoxins in Food Matrices
 
The webinar was hosted by LC/GC Magazine and Thermo Scientific.
 
Live Webcast: Tuesday, August, 14, 2012 at 8:00 am PDT; 11:00 am EDT; 15:00 GMT

Register Free at http://www.chromatographyonline.com/foodmatrices
EVENT OVERVIEW
Mycotoxins are found in a staggering range of matrices, from grains to fruits, spices and nuts to animal feed. They are of major concern for their negative impact on the worldwide economy from product revenue losses, their toxicity to animals and humans, and their persistence throughout multiple food processing steps.Learn how Pickering Laboratories addresses this challenge and meets guidelines set by the FDA using sensitive and rugged Thermo Scientific UHPLC methods to effectively detect Aflatoxins as well as other key classes of Mycotoxins. These easy-to-use methods utilize immunoaffinity clean-up and post-column derivatization.

Also join us to discover the benefits of a Thermo Scientific LC-MS/MS method using automated Turbulent Flow sample extraction to separate Mycotoxins from complex food matrices. A highly efficient solution used in high throughput laboratories that eliminates manual sample preparation, improves method reliability and drives down cost per sample.

Who Should Attend
•
Analytical Chemists
•
Lab Managers
•
Food Science Researchers
Presenters
Wendy E. Rasmussen
Wendy E. Rasmussen
Technical Expert
Food, Agricultural and Environmental Markets
Pickering Laboratories
Markus M. Martin
Markus M. Martin
Solutions Manager LC/MS
Thermo Fisher Scientific
Dr. Yang Shi
Dr. Yang Shi
Senior Applications Scientist
LC/MS Applications
Thermo Fisher Scientific
Laura Bush
Moderator:
Laura Bush
Editorial Director
LCGC North America
Presented by
Sponsored by
LC/GC Logo
Thermo Logo
 
amino acid, Amino Acid Analysis, column, GARD, Guard, HPLC, pickering

New! GARD: Column Protection System

A new Column protection system for Cation-exchange HPLC applications will be unveiled exclusively at Pittcon this year.

The new GARD™ manufactured by Pickering Laboratories is a substantial improvement over the standard packed guards. The new GARD™ adds little pressure, is invisible to the chromatography, and has substantially more capacity for strongly retained compounds that can foul the analytical column.

The new GARD Column Protection System significantly prolongs column life without band spreading or added pressure. We will have a poster at Pittcon demonstrating, by means of a performance comparison for Amino Acid Analysis, that the use of a GARD will protect the analytical column more effectively than traditional guard cartridges, is more cost-effective for the laboratory, is easy to change, and most importantly has zero band spreading.