Category Archives: Amino Acid Analysis

amino acid, Amino Acid Analysis, Chromatography Quiz, Uncategorized

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, Chromatography Quiz, Michael Pickering, Ninhydrin, Trione, Uncategorized

Chromatography Quiz #16

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Chromatography Quiz #15 Results

We would like to congratulate our grand prize winners of our last newsletter’s Amino Acids Analysis Chromatography Quiz: Tom Schneider from Suffolk County Water Authority, Jim Balk from DHHS Public Health Environmental Laboratory, and Narjes Ghafoori from LA County Environmental Toxicology Laboratory!

They have each won and will shortly be receiving: a $100 gift card for Best Buy, which can be used online or in the store!!!

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

The correct answer for the Amino Acids troubleshooting chromatogram: the baseline noise was caused by a dirty flow cell. As we have seen through several Chromatography Quizzes, baseline noise can have a variety of causes, both mechanical and chemical. In this chromatogram, the noise we are seeing is not regular or repeatable, steering us away from the HPLC pump, a trapped bubble, or the back-pressure regulator.

To learn more about baseline noise, please review Maria’s article entitled “The Art of Noise” (http://pickeringlabs-retentiontimes.com/?p=58) from a previous newsletter.

Thank you!

Pickering Labs

Chromatography Quiz #16:

We are doing something a little bit different with this quiz! Identify what’s wrong with the picture below and win a prize! Simply email your answer as well as your full contact information to Rebecca at rlsmith@pickeringlabs.com by July 1st, 2014 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 your submission).

A blast from the past: What is wrong with this picture?!

As you can probably tell from the photo, we’ve dug into Michael’s past for our quiz this quarter! Look carefully and see if you can figure out what’s wrong with this picture! If you have any questions, please feel free to email Rebecca at rlsmith@pickeringlabs.com for further guidance.

Michael_Trione Ad

Amino Acid Analysis, Chromatography Quiz

Chromatograpy Quiz No. 15

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Chromatography Quiz #14 Results

We would like to congratulate our grand prize winners of our last newsletter’s Carbamate/Glyphosate Crossword Puzzle Chromatography Quiz: Jim Balk from DHHS Public Health Environmental Laboratory, Amanda Comando and Dan Raphaely from SCWA, June Black from the Pennsylvania Department of Environmental Protection, Narjes Ghafoori from LA County Environmental Toxicology Laboratory, Helene Lachance from Shur-Gain Nutreco, Holger Franz from Thermo Fisher Scientific, Matthew Hartz from Underwriters Laboratories, Hossein Hajipour from Texas Department of State Health Services, and Daniel Durham from Georgia Department of Natural Resources!!!

rum cake picThey have each won and will shortly be receiving: a Tortuga To Go Suitcase Gift Pack from tortugarumcakes.com!  This “vacation in a box” includes three rum cakes and an 8oz package of coffee, shipped directly from the Cayman Islands!

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

The correct answers for the Carbamate/Glyphosate crossword puzzle:

Across:

1. Pickering column protection system: GARD
6. Heated or Ambient: REACTOR
10. ChlorAC buffer for sample ________: PRESERVATION
11. Over-pressure ________ valve: RELIEF
12. Internal Standard: BDMC
13. Faulty check valves yield a noisy ________: BASELINE
14. Solution to late, broad Glyphosate and AMPA peaks: RESTORE

Down:

1. Glyphosate oxidized by hypochlorite: GLYCINE
2. Pickering Catalog number 3700-2000: THIOFLUOR
3. Reagent one: HYDROLYSIS
4. Fourth peak: METHOMYL
5. Glyphosate herbicide: ROUNDUP
7. _______ detection: FLUORESCENCE
8. o-phthaladehyde: OPA
9. Gradient, water and _______ : METHANOL

Thank you!
Pickering Labs

Chromatography Quiz #15:

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 May 1, 2014 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 – Baseline Noise

Pinnacle PCX post-column instrument is being used, in a traditional HPLC setup as recommended by Pickering Labs.  The chromatogram is expanded to show the baseline noise.  The quiz question is: what is causing this problem?

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

Problem Chromatogram:

problem chromatogram
Please use our website to view any reference Amino Acid chromatogram – the problem for this quiz is not application-specific, so any of our AAA methods could be applicable.  If you have any questions, please don’t hesitate to email Rebecca at rlsmith@pickeringlabs.com.

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.

 

Amino Acid Analysis, AOAC, china, Ninhydrin, pickering laboratories, pinnacle pcx, post-column derivatization, Trione, Whey proteing

Calculation of Whey Protein Fraction in Milk-Based Infant Formula

A method for the calculation of whey protein fraction in milk-based formula products was developed and validated by a group of researchers from Abbott Nutrition, Covance Laboratories and AOAC International. The method is based on calculating the ratio of (Asx + Ala)/(Pro+Phe) determined from the amino acid profile of the hydrolyzed sample.

The method was approved as AOAC Official First Action Method 2012.07 and is appropriate for accessing compliance with whey content requirements of GB 10765-2010 (National Food Safety Standard Infant Formula)*. It is applicable to finished products containing both intact and partially hydrolyzed whey proteins.

According to this method, the Amino Acid Profile of hydrolyzed samples is determined using cation-exchange chromatography with post-column derivatization and UV/Vis detection. The researchers used Pickering Laboratories post-column derivatization system, our patented Trione Ninhydrin reagent and Pickering Sodium analytical column and buffers.

The method performance was evaluated using non-fat dry milk and products with whey levels from 32% to 63%, including NIST Infant Formula Standard Reference Material. The repeatability ranged between 0.3 and 2.5 %, while intermediate precision were between 2.6 and 3.4%. Average recoveries ranged from 97 to 100%.

The unmatched accuracy and precision of amino acid analysis by HPLC with post-column derivatization allowed the researches to develop a robust mathematical model for calculating whey protein fraction that can be successfully applied to a wide range of products. This proved once again that post-column derivatization analysis of amino acids is still a method of choice for laboratories that require highly accurate results.

Pickering Laboratories offers a total solution for amino acids analysis, including post-column derivatization systems, reagents, columns, buffers and standards.

The reference to the published study is below. The First Action Official Method 2012.07 can also be found online at http://www.eoma.aoac.org/.

Calculation of Whey Protein Fraction in Milk-Based Infant Formula: First Action 2012.07

Authors: Wesley Jacobs, Paul Johns, Phillip Haselberger, Joseph J. Thompson, Darryl Sullivan, Steve Baugh.

Source: Journal of AOAC International, Volume 96, No. 3, 2013, pp. 502-507

 

(*) this is a Chinese Standard