Source: OXFORD BIOMEDICAL RESEARCH, INC. submitted to NRP
DEVELOPMENT OF RAPID, POINT-OF-USE DRY CHEMISTRY DIP-STICK ASSAYS FOR FOOD QUALITY.
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1012542
Grant No.
2017-33610-26653
Cumulative Award Amt.
$99,872.00
Proposal No.
2017-00398
Multistate No.
(N/A)
Project Start Date
Jun 1, 2017
Project End Date
May 31, 2018
Grant Year
2017
Program Code
[8.5]- Food Science & Nutrition
Recipient Organization
OXFORD BIOMEDICAL RESEARCH, INC.
4600 GARDNER RD
METAMORA,MI 48455
Performing Department
Research and Development
Non Technical Summary
Oils and fats are essential nutrients for humans, providing fatty acids necessary for good health as well as being concentrated sources of calories. Fats enhance the taste, texture, and mouthfeel of many foods, and are functionally required in preparing a variety of food products. Frying in oil has been used as a cooking method for over 3,500 years, and has remained a popular way to prepare meals and snacks. However, as the populace becomes more educated and aware of the food they consume, fried foods have come under scrutiny due to their fat content and deleterious effects on health. With normal usage, frying oil degrades to form products that affect the taste and texture of the food, ultimately being discarded when it is "spent" and no longer suitable for cooking.We have developed a proprietary colorimetric technology that visually indicates the degradation of frying oil, changing from blue/black to yellow as the amount of total polar compounds (TPC) increases. These compounds are formed during the frying process, with a limit of 25% TPC being adopted in several countries. A high level of TPCs can cause the fried food to cook unevenly, absorb unwanted oil, and has shown to be deleterious to health. The level of TPCs in frying oil is considered to be the best indicator of its lifetime.Currently, the gold standard for measuring total polar compounds in frying fats is the American Oil Chemists' Society Official Method Cd 20-91 "Determination of Polar Compounds in Frying Fats". This is a laboratory-based method which requires special facilities, training, equipment, and takes approximately three hours to perform. Several hand-held conductivity tests have been developed to measure TPC, but are subject to interferences due to differences in types of fats and cost hundreds of dollars.We propose to complete the development of an inexpensive, easy-to-use dipstick prototype that measures % TPC and undergoes a drastic color change indicating when frying oils and fats should be discarded.Our preliminary work has produced a prototype dipstick which gives a very noticeable color change from 0% to 25% TPC. The work performed in Phase I will optimize this color change by evaluating several dye matrices, the effect of oil type, and by modifying the dye formulation. The objective of this work is to develop a convenient, inexpensive, and easy-to-use frying oil test which requires minimal training and undergoes a drastic color change with oils that are at the end of their usable life. This test will offer the fry operator a method for monitoring oil quality to improve taste, health, and alsoGoal 1: Evaluate several commercially-available glass fiber and/or polymer matrices to obtain the most robust formulation that will hold up to handling during manufacturing and also display the maximum color change when used in the assay.Goal 2: Modify the dye formulation, excipients, and synthesize new dye congener.Goal 3 Using a series of new and used oil samples standardized using the AOCS method, produce color charts correlating % TPC to end-point color for four commodity frying fats.Long-Term Objective: Create a suite of point-of-use tests which, with the aid of a portable cell-phone based instrument, measures the freshness and quality of food ingredients using chemical and/or immunosorbent assays converted to dipstick format.
Animal Health Component
30%
Research Effort Categories
Basic
(N/A)
Applied
30%
Developmental
70%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
71118992000100%
Goals / Objectives
Oils and fats are essential nutrients for humans, providing fatty acids necessary for good health as well as being concentrated sources of calories. Fats enhance the taste, texture, and mouthfeel of many foods, and are functionally required in preparing a variety of food products. Frying in oil has been used as a cooking method for over 3,500 years, and has remained a popular way to prepare meals and snacks. However, as the populace becomes more educated and aware of the food they consume, fried foods have come under scrutiny due to their fat content and deleterious effects on health. With normal usage, frying oil degrades to form products that affect the taste and texture of the food, ultimately being discarded when it is "spent" and no longer suitable for cooking.We have developed a proprietary colorimetric technology that visually indicates the degradation of frying oil, changing from blue/black to yellow as the amount of total polar compounds (TPC) increases. These compounds are formed during the frying process, with a limit of 25% TPC being adopted in several countries. A high level of TPCs can cause the fried food to cook unevenly, absorb unwanted oil, and has shown to be deleterious to health. The level of TPCs in frying oil is considered to be the best indicator of its lifetime.Currently, the gold standard for measuring total polar compounds in frying fats is the American Oil Chemists' Society Official Method Cd 20-91 "Determination of Polar Compounds in Frying Fats". This is a laboratory-based method which requires special facilities, training, equipment, and takes approximately three hours to perform. Several hand-held conductivity tests have been developed to measure TPC, but are subject to interferences due to differences in types of fats and cost hundreds of dollars.We propose to complete the development of an inexpensive, easy-to-use dipstick prototype that measures % TPC and undergoes a drastic color change indicating when frying oils and fats should be discarded. Our preliminary work has produced a prototype dipstick which gives a very noticeable color change from 0% to 25% TPC. The work performed in Phase I will optimize this color change by evaluating several dye matrices, the effect of oil type, and by modifying the dye formulation. The objective of this work is to develop a convenient, inexpensive, and easy-to-use frying oil test which requires minimal training and undergoes a drastic color change with oils that are at the end of their usable life. This test will offer the fry operator a method for monitoring oil quality to improve taste, health, and alsoGoal 1: Evaluate several commercially-available glass fiber and/or polymer matrices to obtain the most robust formulation that will hold up to handling during manufacturing and also display the maximum color change when used in the assay.Goal 2: Modify the dye formulation, excipients, and synthesize new dye congener.Goal 3 Using a series of new and used oil samples standardized using the AOCS method, produce color charts correlating % TPC to end-point color for four commodity frying fats.Long-Term Objective: Create a suite of point-of-use tests which, with the aid of a portable cell-phone based instrument, measures the freshness and quality of food ingredients using chemical and/or immunosorbent assays converted to dipstick format.
Project Methods
To complete the development of the current TPC dipstick prototype into a viable commercial product, it is critical to prepare a formulation that is mechanically and chemically robust and capable of consistent performance in spite of environmental differences in real world frying situations. The following Technical Objectives will be addressed during Phase I of this project:Objective 1: Attain maximal pad color development in less than two minutes.The current prototype undergoes a significant color change achieving, in 3-5 minutes a stable pseuo-plateau, with color stability for over an hour afterward. Based on feedback from food suppliers and restaurants, the ideal time from dip to a stable color ready to read is less than two minutes. The rate of color development can be accelerated by adjusting the concentrations of excipient, dye, and matrix pad material and/or adding an additional related congener of one of the present proprietary dyes which will be synthesized in-house.Objective 2: Obtain a non-interfering and visually-homogenous pad matrix.Simple cellulosic test pads have multiple hydroxyl groups that are not suitable for use for TPC testing. The current prototype uses a specific glass fiber matrix that does not have a chemical binder, which can interfere with the assay. However, due to the manufacturing process glass fiber matrices have a "stippled" appearance that can cause uneven absorption of the chemical formulation and thus some unevenness of the color upon exposure to oil samples.Further, glass fiber matrices without binders are more fragile, presenting difficulties for high throughput manufacturing methods, and are also less-than-ideal with regard to the uniformity of the color change. Several non-cellulosic matrices are available which may provide a more uniform color change. Based on our experience with the development of dry chemistry dipsticks involving many different chemistries, we anticipate that other matrices may help eliminate the occasional mottled color changes that we have thus far observed.Objective 3: Correlate AOCS %TPC to test pad color for various oil types.A wide range of plant materials are used as to produce for frying oils (see Figure 5). We have thus far tested our TPC dipstick prototypes on a limited number of frying oils and blends, including canola, commodity soy and high oleic soy oils. Although the selected dye mixture was selected to be responsive to polar functional groups, and has thus far shown a very strong correlation to the AOCS reference method, we recognize the possibility that the test method may exhibit some differences in color end-points or the progression of color change for some different frying oils and/or commercial frying oils or blends. Further, to reduce foaming and improve other characteristics, commercial cooking oils often include additives. These are typically present at parts per million levels, and are not expected to interfere with the color reaction. Nevertheless, it is important to evaluate and address these possibilities, especially for the oils, blends, and additives that are most commonly used for frying foods, Therefore, working with collaborators at Crystal Filtration, DuPont-Pioneer and others, we will obtain fresh oils as well as oils sampled at different stages in the frying process. We will measure the %TPC of each oil type or blend using the AOCS method, and will comparing the color of the test strip to other oils with the same %TPC value.If, for certain types of oils, blends or additives, differences are observed between the rates of change or end point values obtained using the dry chemistry dipstick vs. the AOCS method, then we will make adjustment the formulation to eliminate such variations. If significant differences are observed that cannot be addressed by reformulation, then we will identify groups of oils that perform similarly, and will be able to formulate two or more products for use with groups of oil types.Objective 4: Compare pad color change with various oil types.Differences in fatty acid composition lend diverse properties to each oil type, with coconut being a solid wax at room temperature and soy being a liquid. We hypothesize that these variations may cause different oils with the same TPC content to react differently with the dry chemistry components of our dipstick technology, either with a different color end-point or with a different rate of color change. During this phase of the project, four oils with significantly different physical/chemical properties will be evaluated for the performance of the TPC dipstick. Three calibrators, one new at 0% TPC, the second with 15% TPC and the third at 30% TPC by AOCS will be used, and the color response for each oil will be compared both visually and instrumentally using our Spectrocam® or requested Ocean Optics® reflectance spectrometers.Objective 5: Evaluate shelf life and stability after manufacture and during use:Upon deciding on a final formulation, several hundred dipsticks will be manufactured and placed into opaque plastic vials with oxygen scavengers and desiccants. While a majority of these will be placed at -20C for future use, 20 to 30 vials at 30 strips per vial will be placed at 50?C, 70?C and be measured on a weekly basis with known calibrators. The color response will be measured with the Spectrocam® or Ocean Optics® reflectance spectrometers.Objective 6: Compile data from Objectives 1 thru 4:This task will involve the compilation of the technical data from Tasks 4 and 5 to create a product manufacturing protocol (PMP) and a standard operating procedure (SOP). These two documents will be used to expand the dipstick manufacturing from hundreds at lab bench-scale to small batch runs numbering in the thousands.

Progress 06/01/17 to 05/31/18

Outputs
Target Audience: Nothing Reported Changes/Problems:Problems Encountered and Addressed: A novel dye with a similar structure but was synthesized to determine if its electronic characteristics might improve the color change range of the assay, but it only reacted with low polarity oil and was abandoned. Theproprietary blue dyewhich we have used in the formulation since the project's inception was found to give an improved color change at twice its original concentration, but further increases in concentration did not alter the response appreciably. The oxygen scavenging packet used to store previous lots of finished dipsticks were found to be detrimental to the pads, possibly due to degassing of a reactive component and will be eliminated from future lots. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?Planned Activities: One additional, structurally-similar dye is currently being evaluated to see if it can further enhance the color-change of the pad, and if this is not successful, then the current formulation will focus only on the blue dye. The other additives in the formulation will be evaluated in a similar systematic fashion to find their optimal concentrations. In addition to high oleic acid oils presently being used as calibrators, commodity soy and canola oils are going to be concentrated and calibrator sets prepared to complete the evaluation of four of the most common fry oils.

Impacts
What was accomplished under these goals? Achievements: Project Summary: The total polar compound (TPC) dipstick formulation for assessing frying oils has been modified to improve performance and is near finalization. The test pad matrix material was improved by switching to a different glass fiber paper, while final product storage conditions were improved by removing the oxygen scavenging packet. The dipsticks were found to be sensitive to the oxygen scavenging packet used to store the finished product, and are also sensitive to bright light. A thermal stability study using dipsticks prepared before the beginning of this grant was initiated. Background:The goal of this Phase I project is to finalize the development of a dipstick test that changes color as cooking/frying oil ages and nears the end of its lifetime. The work performed during this grant is a continuation of several years worth of development of a formulation containing two dyes that change color based on the polarity of the solvent that they are exposed to; in this case, used frying oil. The color change of the dyes in the dipstick is proportional to the total polar content (TPC) of the used oil, a test that will help restaurants as well as manufacturers of fried food products determine its discard point. The formulation resulting from this work gives a distinct color change from new and used oils, but the change is not as visually distinct as other similar tests, such as pH strips. The formulation and finished dipstick will be evaluated for storage at elevated temperatures, and tested with various types of oil to determine if a single formulation works for various oil compositions, or requires a different scale for various oil compositions. Process:To optimize the color change of the dipstick pad, the concentrations of the dyes were varied systematically to find their optimal concentrations. These changes were measured both visually and using a reflectance spectrometer. Formulations were prepared which contained only one dye to show its contribution to the overall color change, resulting in the elimination of one of the two dye components from the formulation. A thermal stability study using a pilot lot of dipsticks prepared prior to this grant is on-going and will be described in the final report. Findings and Conclusions:After evaluating several pad materials, the prototype formulation was modified to utilize a glass fiber matrix that is more physically robust than the B-85 material used in previous formulations. The red dye used in all previous formulations will no longer be evaluated as it does not contribute significantly to the overall color-change of the assay. Note: The full report in PDF format will be submitted tosbir@nifa.usda.gov.

Publications


    Progress 06/01/17 to 05/31/18

    Outputs
    Target Audience:The target audience for this project is the restaurant owner, either small or large chain, that fries foods on a daily basis. While the Phase I portion of this project did not produce a final product for sale, we exhibited a semi-final prototypeat the National Restaurant Association meeting in Chicago and received signficant positive feedback from over 150 potential customers ranging from owner/operators to representatives of mutiple restaurant chains in the US and from many other countries. At this meeting, we demonstrated our FryCheck oil test strips and advertized them as "Coming Soon". Changes/Problems:A three-month extension was granted to allow for the completion of the stability study after two final formulations were prepared. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

    Impacts
    What was accomplished under these goals? Project Summary: Proof-of-concept has been established for the dipstick assay for measuring total polar compounds in frying oil. Two formulations have been optimized over the past year of this Phase I grant; one for high oleic- and one for commodity soybean oil. During this time, several key changes were made to obtain two final formulations: the test pad matrix material was changed to a different glass fiber paper which greatly improved its handling properties during manufacturing and after packaging; the dipsticks were found to be sensitive to the oxygen scavenging packet used to store the finished product, which was removed from further study. An accelerated stability study using dipsticks optimized for high oleic soybean oil prepared at the end of the grant were found to maintain their usefulness when stored at room temperature over the course of 50 days. Several oil calibrator sets were prepared from used oils to prepare reference solutions to use for future studies. The following Objectives were addressed and completed: Objective 1: Attain maximal pad color development in less than two minutes. Finalized Formulations for Commodity and High Oleic Soybean Oils. By examining the individual dyes separately in dipstick formulations, it was discovered that the red dye contributed very little to the visual color-change observed when measuring oils with higher total polar compound content than 10%. Also, it was found that the blue dye could be doubled in concentration to give a much more pronounced visual change while simplifying the formulation. A Design of Experiment statistical program was then used to guide us in optimizing the concentration of all formula components to obtain maximal color change. Two formulations were ultimately selected to attain optimal performance for commodity soy and for high oleic soy oils. These formulations will now be taken to the next step of manufacturing (ca. 100,000 dipsticks or more) in beta test lots. Objective 2: Obtain a non-interfering and visually-homogenous pad matrix. Improved Pad Matrix. Initial formulations used a glass fiber paper to make the pad for the dipstick. While very inert towards reacting with the dyes and other excipients in the formulation, the pad material, I.W. Tremont B-85, was very difficult to handle as it was produced without a binder. Hence, multiple matrix materials were evaluated and a glass pad material with superior wet handling properties and inertness towards the assay chemistry, Grade F from the same manufacturer, was identified and selected for all future dipstick work. This matrix material also gave a better color change both visually and using the Ocean Optics reflectometer. Objective 3: Correlate AOCS %TPC to test pad color for various oil types. Oil Calibrator Sets. Our prior studies relied on relatively small samples of fresh and used frying oils for which %TPC was calibrated using the AOCS method. Unfortunately, the small volumes of used oils with known total polar values were quickly exhausted. Therefore, calibrator sets comprised of four samples with total polar values obtained using the AOCS-method were created for high oleic soybean and high oleic canola oils. Using silica gel and a modification of the American Oil Chemists' Society method for determining total polar compounds, two large samples of used frying oil were concentrated and separated into a low and high polarity samples. Together with unused oil and used oil that was not processed, this provided large volumes (ca. 100 mL) of four calibrators with defined TPC content. Visual and Instrumental Comparison of Color Change. The reflectance measurements and visual comparisons of many different variations of the dipstick formulation resulted in two finalized formulation with maximal color change for commodity and high-oleic soybean oil. Objective 4: Compare pad color change with various oil types. Reviewers of our Phase I application stated that the amount of work initially proposed was overly-ambitious. Therefore, in accordance with their suggestions, we focused on a limited the number of different oil types for Phase I studies: high-oleic and commodity soybean oils. Should the Phase II grant application be funded, additional oil and shortening types will be studied in accordance with their commercial usage. Objective 5: Evaluate shelf life and stability after manufacture and during use: Streamlined Dipstick Storage Conditions: A heated stability study showed that dipsticks stored at freezer and room temperatures did not significantly degrade over the course of several weeks. The dipsticks were found to be sensitive to intense white light, mandating the use of opaque vials for future dipstick lots. Using a lot of dipsticks prepared before this grant was awarded, the Ocean Optics reflectometer was used to determine that an oxygen-scavenger packet added to vials containing a TPC dipstick prototype did not promote stability, but caused the formulation to degrade. This resulted in removal of the oxygen-scavenging packet from further lots, and use of only a clay desiccant as a preservative. Objective 6: Compile data from Objectives 1 thru 5: This final report was generated and the Phase II grant application was submitted. Business-Related Achievements The goal of this project is to create a product that will improve the health and safety of fried foods, and create a profitable product for Oxford Biomedical Research (OBR). With the two finalized formulations in hand, OBR launched a new division called Food Quality Testing Corp. to market and distribute our new frying oil tests, which we are branding as FryCheck™. These unique names have been registered with the State of Michigan corporation's division, and a new website has been developed that is focused on food quality testing products. In the middle of May, we will be introducing our initial product, FryCheck™ for commodity and high oleic soybean oils, at the National Restaurant Association in Chicago. Since we still need to manufacture a large production lot with the appropriate quality assurance testing, we will label FryCheck™ as "Coming Soon" and use the opportunity to obtain the voice of many potential customers and - if needed - modify our future design, packaging and marketing efforts to best meet the needs of the marketplace.

    Publications


      Progress 06/01/17 to 01/31/18

      Outputs
      Target Audience: Nothing Reported Changes/Problems:Problems Encountered and Addressed: A novel dye with a similar structure but was synthesized to determine if its electronic characteristics might improve the color change range of the assay, but it only reacted with low polarity oil and was abandoned. Theproprietary blue dyewhich we have used in the formulation since the project's inception was found to give an improved color change at twice its original concentration, but further increases in concentration did not alter the response appreciably. The oxygen scavenging packet used to store previous lots of finished dipsticks were found to be detrimental to the pads, possibly due to degassing of a reactive component and will be eliminated from future lots. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?Planned Activities: One additional, structurally-similar dye is currently being evaluated to see if it can further enhance the color-change of the pad, and if this is not successful, then the current formulation will focus only on the blue dye. The other additives in the formulation will be evaluated in a similar systematic fashion to find their optimal concentrations. In addition to high oleic acid oils presently being used as calibrators, commodity soy and canola oils are going to be concentrated and calibrator sets prepared to complete the evaluation of four of the most common fry oils.

      Impacts
      What was accomplished under these goals? Achievements: Project Summary: The total polar compound (TPC) dipstick formulation for assessing frying oils has been modified to improve performance and is near finalization. The test pad matrix material was improved by switching to a different glass fiber paper, while final product storage conditions were improved by removing the oxygen scavenging packet. The dipsticks were found to be sensitive to the oxygen scavenging packet used to store the finished product, and are also sensitive to bright light. A thermal stability study using dipsticks prepared before the beginning of this grant was initiated. Background:The goal of this Phase I project is to finalize the development of a dipstick test that changes color as cooking/frying oil ages and nears the end of its lifetime. The work performed during this grant is a continuation of several years worth of development of a formulation containing two dyes that change color based on the polarity of the solvent that they are exposed to; in this case, used frying oil. The color change of the dyes in the dipstick is proportional to the total polar content (TPC) of the used oil, a test that will help restaurants as well as manufacturers of fried food products determine its discard point. The formulation resulting from this work gives a distinct color change from new and used oils, but the change is not as visually distinct as other similar tests, such as pH strips. The formulation and finished dipstick will be evaluated for storage at elevated temperatures, and tested with various types of oil to determine if a single formulation works for various oil compositions, or requires a different scale for various oil compositions. Process:To optimize the color change of the dipstick pad, the concentrations of the dyes were varied systematically to find their optimal concentrations. These changes were measured both visually and using a reflectance spectrometer. Formulations were prepared which contained only one dye to show its contribution to the overall color change, resulting in the elimination of one of the two dye components from the formulation. A thermal stability study using a pilot lot of dipsticks prepared prior to this grant is on-going and will be described in the final report. Findings and Conclusions:After evaluating several pad materials, the prototype formulation was modified to utilize a glass fiber matrix that is more physically robust than the B-85 material used in previous formulations. The red dye used in all previous formulations will no longer be evaluated as it does not contribute significantly to the overall color-change of the assay. Note: The full report in PDF format will be submitted tosbir@nifa.usda.gov.

      Publications