We will be displaying our equipment at this year’s Pittcon in Chicago, Illinois from March 3-6, 2014. Be sure to stop by: Booth 2955
We will have our Pinnacle PCX post-column derivatization instrument on display as well as our Mycotoxin Analysis Products. In addition, we will some new videos to highlight the latest in our Automated Sample Preparation instrument the FREESTYLE SPE system.
The new FREESTYLE system is the most adaptable robotic equipment for sample preparation currently available on the market. With its nearly universally applicable system range, numerous processing steps can be elegantly automated. The FREESTYLE system takes on daily routine tasks in the laboratory, but also offers the user the unique opportunity to combine specific working steps that were previously carried out individually.
FREESTYLE SPE Module: The FREESTYLE SPE system is suitable for the fully automated processing of the most diverse column formats. The technology is unique:
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FREESTYLE EVAporation Module:
The FREESTYLE EVAporation system is suitable for the fully automated evaporation of many samples – if you wish overnight or over the weekend. The novel FREESTYLE technology:
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FREESTYLE GPC Module: The FREESTYLE GPC module facilitates the upgrade to a fully automated chromatographic sample preparation system (gel permeation chromatography), as is used for example for clean-up of food, animal feed and environmental samples. It complies in all areas with the requirements of general methods, e.g. DFG S19, EPA 3640A, AOAC 984.21, EN 1528 and more. The module consists of a preparative LC-double piston pump with all its advantages e.g. minimizing pulsation. With a flow rate of say 5 mL/min, the preparative pump incurs only a small load and is consequently particularly low maintenance.
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Be sure to visit www.freestyle-robotic.com for more details, configurator, and applications
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.
Determining 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 Conditions
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
The 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 
toxin 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
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:
The brochure for ThermELUTE can be found HERE. To view the slides from our presentation, click HERE.
A Story with Bearing: Cholesterol
By Michael Pickering
Years ago our friend Peggy told us that her doctor had prescribed a diet devoid in cholesterol, as her blood test indicated worrisome numbers. The doctor recommended that all of the usual suspects be excluded from her diet, such as egg yolks and butter, but it was all to no avail. Regardless of how long she maintained the exclusive diet, her blood numbers did not budge. So, she decided to experiment with her own dietary exclusions. One of her first experiments was targeted at a food everyone who knows Peggy is well aware that she is addicted to – milk chocolate. It was only then that her blood cholesterol numbers improved.
While it is true that one’s diet is an important factor in the level of cholesterol in one’s blood, the amount of cholesterol in one’s diet is not germane. Unlike the essential amino acids and minerals which must be harvested from the diet, the cholesterol in our blood is synthesized inside our bodies from smaller synthons (many acetates, a popular biosynthetic mode, see flavonoids).
So the issue isn’t whether cholesterol is in one’s diet, but rather how cholesterol is behaving in one’s blood.
The key link between the importance of diet and the behavior of cholesterol in one’s blood is the amount and type of fat you ingest. Highly saturated fats have the most negative impact on the solubility of cholesterol in the blood.
Blood chemistry is necessarily dominated by water soluble processes. Magnesium, sodium, citrate, and all manner of water soluble nutrients must course around freely. However, cholesterol is not water soluble, even though it must move as freely through our veins. The body’s solution is to cloak the cholesterol with a hydrophobic interior (cozy coat) with a hydrophilic exterior (sort of like a 1960’s Bill Blass coat with the mink on the inside and the satin on the outside).
These water taxis are called LDL and HDL: low density lipoproteins and high density lipoproteins. Our body considers the HDL to be better than the LDL because, among other things, it’s easier to void. Tom Scheve’s description of the reason for this in his article for Discovery Health, entitled “What’s the difference between LDL and HDL Cholesterol” eloquently expresses my own musings:
When the lipoprotein has more protein than cholesterol [HDL], it resembles a Ferrari, gunning through your body without stopping until the cholesterol arrives at your liver, where it’s converted into bile acids. […] When the lipoprotein has more cholesterol than protein [LDL], however, this makes for a rickety ride, and that jalopy doesn’t get too far. Cells have special receptors that bind tightly to these lipoproteins as they pass. This LDL sputters down the road, careening off the arteries, running into things and leaving bits all over the place. While the HDL Ferrari sees a pileup and nimbly speeds around it, the LDL jalopy crashes right into it, adding to the jumble of tangled fenders and tailpipes (or platelets and plaque).
The overall solubility of cholesterol in the blood is governed by a ternary phase diagram. 
Maintaining these three components in the proper ratio crates a zone of solubility in the triangle. If the diet (the source of phospholipids and fats) biases the ratio out of the soluble zone, the cholesterol precipitates with the fenders and tailpipes. And like all solids in a moving fluid, they deposit in the zones of slowest flow. In a vascular system the slowest flow is in the arteries.
The lipids (fats) in our diet can be broadly sorted into two categories: 1) naturally occurring, and 2) man-made. Obviously the naturally occurring fats and oils are derived from plants and animals. The man-made fats are partially hydrogenated vegetable oils. The saturation level determines the melting point and viscosity regardless of the source. So highly unsaturated lipids like sesame oil have a low melting point and viscosity and so are inappropriate for frying, whereas poly-saturated lard and butter have a high melting point and viscosity, and are well suited for frying. Similarly, an award-winning pie crust can be made with lard or butter, but not with unsaturated oil. Partial hydrogenation thus controls the melting point of the fat and establishes its suitability for any particular application.
However, a side reaction also occurs during the hydrogenation: isomerization. Natural unsaturation tends to be cis-configuration but hydrogenation isomerizes the bonds to trans-configuration.
While the hydrogenation controls the melting point precisely (which is essential for processed foods), the resulting fat is not recognized by the body as food.
Our wild type diet is clearly designed around whole grains as the staple, a source uniquely rich in unsaturated (cis-) fats, phospholipids, and protein. The goal is to manage the trace chemistry in our blood, the hydrophobic components. So the lesson is: eat as little saturated (mostly animal) fat as you can tolerate, eat whole grains and exclude all partially hydrogenated vegetable oil. Read the label!
So while the amount of cholesterol present in a milk chocolate bar (24 g) is comparable to that of a tablespoon of butter (30 g), the two had very different effects on Peggy’s blood work. Cocoa fat is among the most saturated of the vegetable oils. The melting point is so high that the bars are wax-like at room temperature. The chocolate was in effect creating an excess of LDL jalopy wreckage in Peggy’s blood stream, by causing the LDL and HDL levels to get out of whack.
The cholesterol had no bearing.
Editor’s Note: Always consult a physician first. The views presented herein are strictly editorial in nature.
Chromatography Quiz #13 Results
We would like to congratulate our grand prize winners of our last newsletter’s Carbamate Analysis Chromatography Quiz: Irene Taylor from Orange County Utilities, Jim Balk from DHHS Public Health Environmental Laboratory, Narjes Ghafoori from LA County Environmental Toxicology Laboratory, Helene Lachance from Shur-Gain Nutreco, and Jamie Palmer and Matthew Hartz from Underwriters Laboratories!!!
They have each won and will be receiving: a Laser Board Game from 
Sharper Image! (This strategy-based board game is similar to chess and features an 80-square Egyptian themed grid. The goal is to protect your game pieces while eliminating your opponent’s by bouncing eye-safe lasers at them through the maze of mirrors you’ve constructed.)
We would like to thank all of you for your submissions!
The correct answer for the modified Carbamate chromatogram: Reagent #2 (CB910, OPA, and Thiofluor) was partially oxidized. The peak heights of Carbaryl and 1-Naphthol have significant diagnostic importance in regard to the OPA reagent system. Since 1-Naphthol naturally fluoresces, we can eliminate the fluorescence detector as the problem. Though smaller peak sizes can reflect problems with Reagent #1, since the Carbaryl peak is full-sized, we can deduce that hydrolysis has occurred (Carbaryl naturally fluoresces after hydrolysis).
Particularly astute observation yields additional details about our troubleshooting chromatogram – slightly shifting retention times towards the end of the run could indicate a problem with HPLC hardware, such as a proportioning valve.
Thank you!
Pickering Labs
Chromatography Quiz #14:
Complete the cross word puzzle below and win a prize! Simply email your answer (copy of completed puzzle or list answers with clues) as well as your full contact information to Rebecca at rlsmith@pickeringlabs.com by December 15, 2013 in order to win. You will receive email confirmation that your submission has been received. The answer to the puzzle and winner congratulations will be published in the next issue (to be anonymous, please notify Rebecca in submission).
Carbamate/Glyphosate Analysis: Cross Word Puzzle
By Wendy Rasmussen
Before the America’s Cup came to San Francisco, I never paid much attention to the event. It doesn’t get much press here in the US, and being a native of a land-locked state, sailing was not something I grew up with.
But after living in San Francisco for many years, I learned to sail and gradually my awareness of the America’s Cup increased, and I gained an appreciation of the event and the hard work that goes into designing and sailing the boats.
Those who wish to race for the cup become Challengers. If multiple teams are challenging, they must duel it out to decide who will then face the Defender in the America’s Cup Finals. This series of races became known as the Luis Vuitton Cup. From the America’s Cup Website:
(Be sure to check out www.americascup.com for information, videos, history, race results and more).
This year, Emirates Team New Zealand (ETNZ) 
won the Luis Vuitton Cup to become the Challenger, sailing against the Defender, Oracle Team USA (OTUSA).
Living in San Francisco, I had the wonderful opportunity to watch several of the races, to meet many of the sailors, and to watch the teams ready themselves for the “big day”. The teams were in town for several months before the regattas to practice. The AC72s were (are) an amazing site to see, both from land and from the water.
They are truly a sight to behold, especially when foiling at speeds of up to 45kts*
The 34th America’s Cup finals proved to be the longest in history due to several postponements due to wind conditions. In fact, the official end date for the regatta was September 21; the final race was on September 25th. But perhaps the most amazing feat of this regatta was the comeback made by OTUSA:
In order to win the Cup, a team must earn nine points. Ordinarily, this would mean winning 9 races (1pt per win). But before the regatta began, Oracle were penalized two points due to an infraction in an earlier series. And so while ETNZ needed to win 9 races, OTUSA had to 11.
The comeback began on Sept 19, 2013. ETNZ were ONE race away from winning the Cup. OTUSA still need to win EIGHT. For days on end, we all said, “Well, this is it. Kiwi’s are going to take home the cup.” But then OTUSA continuously surprised us all. In the end, they won an unprecedented EIGHT RACES IN A ROW! in order to come back and win the regatta. Click here for the Results.
I won’t speculate here as to why OTUSA made such an amazing comeback, or why ETNZ lost in the end. I will only say that it was so exiting to be a part of that historic moment, in an amazing city with an amazing natural venue for watching the races. And most importantly for me, the members of both teams handled themselves with an incredible amount of dignity and sportsmanship. The members of both teams were all under an incredible amount of pressure, and yet they still managed to greet and sign a few autographs for their fans — yes, this fan included.
*45kts = 51.8mph. By comparison, our sailboat at it’s fastest will do about 12kts, or 13.8mph
