Category Archives: amino acid

Chromatography Quiz #29

Chromatography Quiz #28: 35th Anniversary Word Puzzle – Results

Pickering Labs would like to congratulate all of our winners for our previous newsletter’s 35th Anniversary Word Puzzle: Jim Balk from Nebraska DHHS Public Health Environmental Laboratory, Tom Schneider from Suffolk County Water Authority, Karissa Scroggins from North Coast Laboratories, Narjes Ghafoori from LA County Environmental Toxicology Lab, Joy Gottlieb from New Mexico Department of Health Scientific Lab Division, Hossein Hajipour from Texas Dept. of State Health Services Laboratories, and Widchuda Meeim from Thailand Bureau of Quality Control of Livestock Products.

Winners will soon receive a Packing Organizer Set from the Container Store! This colorful set of 6 packing cubes are durable and clearly labeled to help you organize your suitcase! Just in time as you plan for those summer getaways!

Congratulations to our quiz winners and happy packing!

Thank you all for your submissions! 


             

The correct answers for the Anniversary Word Puzzle are as follows:

Chromatography Quiz #29 – Amino Acids Elevated Baseline:

Simply email your answer as well as your full contact information to Rebecca at rlsmith@pickeringlabs.com by July 15, 2018 in order to win.  You will receive email confirmation that your submission has been received.  The answer to the quiz and winner congratulations will be published in the next issue (to be anonymous, please notify Rebecca in submission). 

Maroon: Good chromatogram

Blue: Elevated baseline

What could be contributing to the elevated baseline?

   

   

   

Chromatography Quiz #21

Chromatography Quiz #20 Results

We would like to congratulate our grand prize winners of our last newsletter’s word problem quiz: Helene Lachance from Shur-Gain Nutreco, Narjes Ghafoori from LA County Environmental Toxicology Lab, and Tom Schneider from Suffolk County Water Authority!  

They have each won and will shortly be receiving: Whimsical Turkey Jumbo Caramel Apple Gift Sets from Mrs. Prindables Gourmet Caramel Apples!

prindables-gourmet-caramel-applesFrom Mrs. Prindables website, “Two fine apples come topped with a fair pair of fowl… sure to be gobbled up with delight. One Triple Chocolate Jumbo apple and one Milk Chocolate Walnut Pecan Jumbo apple topped with our whimsical new turkey ornaments. Make them the centerpiece of your Thanksgiving celebration. Sure to brighten your Fall gathering! Each apple weighs Approx. 1.25-1.5 lbs. Serves 8-10.

Additionally, we will be sending our winners a $100 gift card for Williams-Sonoma to aid in their Thanksgiving Day preparations!

We would like to thank all of you for your submissions! 

Thank you! 

Pickering Labs 

 
  

The correct answers to the Michael Pickering Scavenger Hunt: 

 Question One:

Through what organization did Michael first meet Laszlo Torma, our recently retired Director of Technical Relations?

            Answer: AOAC

Question Two:

To whom is the famous quote “Chance favors the prepared mind” attributed?

            Answer: Louis Pasteur

Question Three:

How long was Michael’s train ride from Anchorage to Fairbanks?

            Answer: Eight hours

Question Four:

What phthalate caused Michael so much consternation both during school and later in a customer’s laboratory?

            Answer: Di-octylphthalate

Question Five:

Michael suggests recipe using saffron – what is he cooking?

            Answer: Poached white fish with saffron infused lime sauce
  

Chromatography Quiz #20: Amino Acids Analysis

Identify the error made when running the Amino Acids chromatogram below and win a prize!  Simply email your answer as well as your full contact information to Rebecca at rlsmith@pickeringlabs.com by December 15th, 2015 in order to win.  You will receive email confirmation that your submission has been received.  The answer to the quiz and winner congratulations will be published in the next issue (to be anonymous, please notify Rebecca in submission). 

Amino Acid Analysis – Reduced Peak Areas

Pinnacle PCX post-column instrument is being used, in a traditional HPLC setup as recommended by Pickering Laboratories.  The reference chromatogram and troubleshooting chromatogram are both shown.  The quiz question: what is causing the reduced peak areas problem? 

Hint: Please assume the same Trione reagent is being used for both chromatograms.

Normal/reference post-column conditions for amino acid analysis:
Reagent: Trione
Reactor: 130 °C, 0.5 mL
Reagent flow rate: 0.3 mL/min

Detection: UV-Vis Detector   = 570 nm for primary amino acids,  = 440 nm for secondary amino acids

quiz-21a
  
 

quiz-21b
  

Amino Acid Analysis of Cell Culture Media

We have a new Method Abstract! MA371, Amino Acid Analysis of Cell Culture Media

Cell cultures are widely used to produce biopharmaceuticals and other biologically active compounds. The composition of the cell culture media affects the yield and structure of the desired products and must be carefully optimized. Cell culture media is typically composed of mixtures of amino acids, vitamins, carbohydrates, inorganic salts as well as different peptides, proteins and other compounds. As the cells grow, they consume nutrients and release target biopharmaceuticals as well as waste products.

Amino Acids serve as the building blocks of proteins, as well as intermediates in many metabolic pathways. Amino Acids are typically added to cell culture media to provide nutritional requirements for the cells. Monitoring and adjusting Amino Acid composition is an essential part of optimizing the manufacturing process to ensure high quality and optimum yield of the final product.

Amino Acid Analysis using cation-exchange chromatography with post-column Ninhydrin derivatization allows for easy determination of Amino Acid concentrations in many complex matrices, including cell culture media. The post-column method is very sensitive, reproducible and rugged. It has been and continues to be a method of choice for laboratories running biological samples, protein, peptides and foods analysis. Most chemical compounds present in the media do not interfere with analysis, so the majority of samples only need diluting with citric buffer and filtering before analysis. If serum is added to the media, then the proteins need to be precipitated using either Seraprep™ solution or ultrafiltration.

Pickering Laboratories, Inc. offers the complete solution for Amino Acid Analysis, including post-column derivatization instruments, columns, eluants, reagents and standards. The Pinnacle PCX derivatization system has a programmable column oven to allow for shorter run times and easy method optimization.

YChromatograms of Cell Culture Mediaou can download this application note, and many others from our website: www.pickeringlabs.com

METHOD
Analytical conditions
Column: High-efficiency Lithium cation-exchange column, 4.6 x 75 mm, Catalog Number 0354675T
Flow Rate: 0.55 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

gradient table for AAA

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/

Chromatography Quiz #9

Chromatography Quiz #8 Results
We would like to congratulate the grand prize winners of our last newsletter’s Carbamates Analysis Chromatography Quiz: Mary Benzinger from North Coast Laboratories, and Keena Njoroge and Matthew Hartz from Underwriters Laboratories!!!
They have won, and will shortly be receiving: Wine Hourglasses courtesy of Uncommongoods.com! Additionally, all participants will be receiving a $20 gift card from Jamba Juice!  Again, we would like to thank all of you for your submissions.  

The correct answer for the modified Carbamates chromatogram: the noisy baseline is due to the HPLC pump having excessively large pulsations (or pressure variations).  Post-column systems containing reciprocating pumps can also cause noisy pulsations in the baseline, and the frequency of the sine waves should be evaluated to determine which pump is causing the noise.

This problem is most often caused by one of the following: bad pump seals, bad check valves, scratched or damaged pump pistons, faulty pulse dampener, or problems with connections to the pump. 

This problem can also be caused by dirty reagents or eluents, however in this case the pulsations would be accompanied by the elevation of the background signal. 

We had a lot of excellent responses to this quiz! 

Thank you! 
Pickering Labs

Chromatography Quiz #9:

Identify the error made when running the Amino Acids chromatogram below and win a prize!  Simply email your answer as well as your full contact information to Rebecca at rlsmith@pickeringlabs.comby May 15th in order to win.  You will receive email confirmation that your submission has been received.  The troubleshooting answer and winner congratulations will be published in the next issue (to be anonymous, please notify Rebecca in submission). 

Amino Acid Analysis of Physiological Fluid

Pickering Standard: 1700-0180 Native Sample Standard, acidics and neutrals, in 0.1 N HCl, 2.5 µmole/mL, 10 µL injection

Pickering Column: 0354100T High Efficiency Lithium Cation-exchange Column, 4.0 x 100 mm

Normal Operating Conditions: (for reference only, condition changes may be reflected in chromatogram)
Column Temperature: 36 °C

Flow rate: 0.35 mL/min

Eluent Gradient:

TIME
Li275 %
Li750 %
RG003 %
0
100
0
0
12
100
0
0
48
65
35
0
90
0
100
0
95
0
100
0
120
0
94
6
122
0
94
6
122.1
100
0
0
140
100
0
0
Post-column conditions for amino acid analysis:
Reagent 1: Trione
Reactor 1: 130 °C, 0.5 mL
Reagent flow rate: 0.3 mL/min

Detection: UV-Vis Detector: 570 nm for primary amino acids,  440 nm for secondary amino acids
An example of a good chromatogram can be found here: Standard Lithium Chromatogram

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.