Research
This web pg is under construction! 7/5/15
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PROFILING OF CONTAMINATES IN DOG FOODS USING ACCURATE MASS TECHNOLOGY AND THE EFFECTS ON DOGS

In 2014 Sue D'Antonio, employee of Agilent Technologies TX, purchased a Red Standard Poodle puppy "Ranger" from me. Her interest in my dog food research awakened her interest to bring it further to her company, and group, due to her job in chromatography (see glossary for definitions).

Every year chromatography companies come together with projects they have chosen to work within their companies/groups to place into an annual contest to further research and support their companies and careers. Among an average 1,000 entries in their industry, 10 are picked for posters with their annual chosen (pet ;o) project.  The research here on dog food contamination is by Agilent Technologies team co-authored by Sue D'Antonio and Tes Ingebritson using Agilent's chromatography equipment.

My own personal and dog food research was the starting story to the poster. These individuals are PHD's in their fields, I can never match that, so here I do my best to bring the information to you in hopeful easy language. A dictionary, and google, at hand will certainly help.

I provided my information to Sue where she worked with her group to test/evaluate over 12 different dog foods. Among the 1,000 entries we were nominated for the poster, the top 10! I can't create a good visual of the poster for you to see it in it's whole, so I will be breaking it down for you on this page with the written information.


Please have a dictionary at hand to help along.

Glossary: 

*(starred) is my added discussion beyond the poster

Chromotagraphy; the separation of mixtures into their constituents by preferential adsorption by a solid, as a column of silica (column chromatography) or a strip of filter paper (paper chromatography) or by a gel.

*Chromotagraphy equipment divides products, foods, etc. to know what is found in them by separating its composition. You will find all major businesses owning one of these machines to know what is found in their products. 

*In working for Agilent, Sue requested to do research in dog food contamination with the companies equipment. THIS WAS PHENOMENAL to help see what is happening to dog foods. Their team broke down over 12 various dry dog food brands found on this page to evaluate what contaminates would be found. Read on to know the conclusion...

Ions; an electrically charged atom or group of atoms formed by the loss or gain of one or more electrons, as a cation (positive ion) which is created by electron loss and is attracted to the cathode in electrolysis, or as an anion (negative ion) which is created by an electron gain and is attracted to the anode. The valence of an ion is equal to the number of electrons lost or gained and is indicated by a plus sign for cations and a minus sign for anions, thus: Na +, Cl−, Ca ++, S =.



Poster Data (working on figure inclusion)

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Introduction

by Tes Ingebritson

Snap, Crackle, Pop! Some dog foods snap life from our pets! As a standard poodle breeder since 2004, I receive communication from around the world on how a pet died from cancer, lymphoma, and ailments like those inflicting mankind. I crackle from grief stricken owners, and then begin research asking what kind of dog food they fed their pet or chemicals used in the environment. Research continued after my husband died in 2014 from cancer. How can we improve our health along with our pets? I had begun eating raw organic foods years before his passing. Feeding my poodles treats of raw organic apple bits, carrots and vegetables et al became a part of their diet.

My viewpoint looked to finding a grain free dog food. The understanding is by eliminating concentrated chemicals in our pet’s food (most are dry) we can decrease the toxins. How? By subtracting grains, which is compounded by grains being ingested by the chickens, beef, et al (the meats) would certainly help. Professors at canine symposiums and seminars shared grains are bulk for dog foods, viewpoints of having benefits, some no benefit. In nature, canines don’t eat grains, so why does man include them? Thus in selling A Perfect World standard poodle at RoyalStandardPoodles.com, I ask customers to research and find a dog food that is grain free, or the healthiest possible food for their pet that causes no ear infections and little flatulence. I have some poodles on food with grains as they get ear infections from grain free. Yet most of my poodles are healthy on grain free. We LOVE our pets! It is hopeful the research found here will help pop awareness into what is in pet foods. Let us snap, crackle, and pop to make a difference in how we feed our pets!

*I have moved the conclusion here that one can capsule as much as possible before taking in the details which I am working on the figures to hopefully be able to bring on.

Conclusions

Conclusion by Tes Ingebritson:

In research to understand what happens to the plant, human or animal body when pesticides, chemicals, and drugs are ingested, digested, or assimilated, this testing shows toxins do not flush out of an organism. It stays in the cells of that organism, then continuing to be ingested, digested, assimilated into the next plant, animal or human, compounding as it cycles through organisms.

The local farmer who allowed petroleum to be knifed into his field because he was paid well to do so, finds his crops dwarf compared to his neighbors. Unable to grow, showing definite plant health issues. What happens then to the animals that feed off of that land? The results can be found in this research. The more organic or holistic has “fewer” toxins in them, yet, are there.

The organic/holistic products here, are still unable to be clear of any toxins due to the fact one can never eliminate, in this day and age, the variables of what happens to a farmers crops, an animals health care from the vet, even though a manufacturer sees to do his best, the ramifications spiral from ignorance to the science that we now know can show irrefutably THE TRANSFERENCE DOES OCCUR IN FOODS AND IS NOT ABLE TO BE ELIMINATED from the food chain! 

 *It is important we all do our best to research the dog food we choose to feed our pets. I have been honored by Agilent's Sue D'Antonio and group for the work they have done on this, free to help us become aware...and their research doesn't stop here...it continues in areas that they are able to further investigate...

a HUGE THANK YOU!

This website helps us evaluate the many dog foods out there:

 http://www.dogfoodadvisor.com/

Agilent Conclusion by Sue D'Antonio:

The Agilent Time of Flight MS combined with our unique software tools allows for confirmation by accurate mass, chromatographic retention time, isotopic spacing and pattern.  In addition the All Ions methodology, adds compound confirmation using fragmentation patterns. This allows us to eliminate false positives in screening methods. We are able to use existing MS/MS data bases for compound confirmation. In addition, the Time of Flight with the fast scan speed and full spectra  is the MS most compatible with UHPLC  fast chromatography.

  This was a qualitative  screening experiment for pesticides and vet drugs. Further quantitative work will be needed to determine actual levels.                  

                                  

 

Results and Discussion

All Ions Concept

The concept of All Ions was used in this series of experiments. Four Voltages were the used in this experiment. The red circle in figure 1A represents where the fragmentor voltage is created. Fragmentor voltage is a value which is comprised  of the voltages at the end of the capillary and the skimmer. These voltage produce no fragmentation of the ions at low voltage and varying fragmentation at higher voltages. This means lower values will show the precursor ion and high values will give product ions using Time of Flight instruments compounds are typically confirmed in four ways. These four confirmations are accurate mass, chromatographic retention time, isotopic abundance and pattern. By using all Ions methodology, a MS/MS like spectral pattern is generated. With this spectral pattern it is possible to search spectral data bases for pattern matching.

70, 120, 170 & 270 were the voltages used in this experiment.

To come with hopeful translation to internet pictures:

Figure 1A, Figure 2, Figure 3, Figure 4a-b, Figure 5a-b, Figure 6a, Figure 6b, Figure 7a-d, Figure 8a-d

Experimental

Thirteen different types of dog foods were analyzed from different suppliers.  All samples were extracted with the QuEChERS protocol.   An Agilent standard containing 14 common pesticides was utilized.  All samples and the pesticide standard were analyzed (n=3) using reversed phase liquid chromatography coupled to a time-of-flight mass spectrometer G6230B.

Figure 3 data:

Sample dog food (see brands used below)- Grain Free - Grain if present - notes - USA - stars

1 - yes - none - sweet potato, chicken meal, chicken fat,chicken - not given - 5

2 - yes - none - real chicken and sweet patato recipe - USA - 5

3 - no - brown rice - Chicken Brown Rice Flax Seed Flavor - USA - 5

4 - no - corn, whole grain sorghum, - chicken - not given - 3

5 - no - barley,oatmeal,brown rice, rye - deboned chicken and oatmeal recipe, tomato - not given - 5

6 - yes - none - chicken recipe - not given - 5

7 - no - corn, soy wheat - Chicken flavor - USA - 0

8 - no - yellow corn, rice, wheat - pork and bone meal preserved poultry fat - USA - 0

9 - no - yellow corn,soy,rice - meat and bone meal, poultry by product meal - not given - 0

10 - no - yellow corn, soy, wheat, rice - chicken, meat and bone meal, - not given - 4

11 - yes - none - cage free chickens and chickpeas, pumpkins and fruits - USA - 5

12 - yes - none - USA - 5

13 - yes - none - USA - 5


Samples List Name Brands used (chicken ingredients focus) that rated 5 star are posted:

1 Natural Balance 

2 Merrick

3 Organix

5 Wellness 

6 Blue Wilderness

8   

10 

11 Dogs well Live Free Dry Dog Foods

12 Health Extension Vets Choice

13 Life's Abundance


In addition, a sample was spiked with the pesticide standard to verify there were no matrix effects.  The data was processed using the Mass Hunter software algorithm capable of reducing the data set into unique compounds found throughout the chromatographic run.  The list of compounds was then filtered by the Agilent PCC database. All the chromatographic conditions, optimized  ToF settings, data analysis method and the Pesticide data base were from the Agilent G6854AA kit.


*Daniel and I went to speak May 6, 2015 at the Minneapolis Chromatography conference with the poster on display. We were welcomed with an astounding filled room to speak to on the results. Daniel came with, and of course, was a big hit accommodating all the petting and acknowledgements of his presence :o)