Recipient Organization
CRAFT TECHNOLOGIES, INC.
4344 FRANK PRICE CHURCH RD.
WILSON,NC 27893
Performing Department
(N/A)
Non Technical Summary
Internationally there are 250 million children whose health is compromised in varying degrees due to the deficiency of vitamin A (VAD) in their diets. One of the most accepted methods for sustained remediation of VAD has been through food fortification. Currently more than 40 countries have implemented VA fortification of foods. The effectiveness of fortification programs and the stability of fortified foods in various environments must be measured. If this does not occur the VA may not be added uniformly or at all; or it may become inactive before the food is consumed. There is a need for a rapid economical means to measure the fortification of foods with vitamin A. The purpose of this project is to develop a rapid field-test (Craft Rapid vitamin A Field Test) to measure vitamin A in four types of fortified foods (sugar, rice, flour, and dairy products) based on the use of a modified portable fluorometer, the CRAFTi.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Goals / Objectives
The overall objective for this proposal is to prepare the CRAFTi, a portable fluorometer, for commercial production including finalized extraction methods and kits for fortified sugar, rice, flour, and milk. This will include accessories, packaging and a marketing/distribution plan. Specific objectives for this proposal include: 1)Refined modifications to the CRAFTi fluorometer. a)Evaluate type and source of lamps, power supply and filters to insure ease of manufacture. b)Test the utility of multiple power options including solar, AC, and DC. c)Modify software or install shutter to eliminate the flaring due to stray light. d)Modify software to test correctly during instrument initialization. 2)Finalization of extraction methods for sugar, rice, flour. a)Finalize sugar extraction using selected solvents and measuring utensils. b)Refine rice extraction and test with more replicates of actual samples. c)Refine flour extraction and test with more replicates of actual
samples. d)Refine corn meal extraction and test with more replicates of actual samples. 3)Development of an extraction method for milk. Spend concerted effort to extract VA from milk without the presence of fluorescent interferences. 4)Analyze at least 25 samples of each matrix obtained from countries fortifying that matrix for statistical comparison of CRAFTi results to HPLC results 5)For each matrix, to analyze the same sample ten consecutive times on the same day using the CRAFTi to determine within-day reproducibility. For each matrix, to analyze the same sample in duplicate on ten consecutive working days using the CRAFTi to determine between-day reproducibility. 6)If the power supply should be modified, recertification of instrument safety using the altered power supply. 7)Identification/development of accessories for measuring, mixing etc. 8)Identification of vendors to supply parts for the instrument modification. 9)Identification of vendors for the solvents and consumable
kits. 10)Identification of packaging materials and marketing strategies. 11)Production of documentation in the form of written manuals and training videos.
Project Methods
Instrument Modification An instrument measuring 6" x 13" x 5" and weighing less than 15 pounds has been modified and laboratory-tested for the measurement of VA. During a field-test, exposure of the sample compartment to bright sunlight resulted in oversaturation of the diodes. We propose to correct this problem by modifying the software or introducing a shutter to block light from the optics. The instrument must be operable with a rechargeable battery power supply. Present modifications have made it operable using a 12-volt battery in addition to AC current. Since the original power supply automatically senses 110 or 220 V AC, it is capable of being used worldwide for AC operation. We will test the use of a voltage inverter that converts DC to AC current. The inverter would only be used during battery operation. Field-Friendly Extractions Extracts of each matrix will be prepared using saponification and measured by HPLC and the CRAFTi. A portion of the FF extract
measured on the CRAFTi will be analyzed by HPLC. Using this comparison between methods and sample preparation, we will be able to choose the approach with the highest extraction efficiency. The measurement of FF extracts using HPLC with fluorescent detection will allow us to determine if fluorescent interferences contribute to the CRAFTi readings. If food matrix components are found to interfere, methods to exclude the matrix fluorescence will be developed. The samples may require filtration or solid phase extraction (SPE) to exclude interfering components. These can both be accomplished using disposable syringes and filters or SPE discs. Sample preparation should be designed to eliminate refrigeration and centrifugation. We have identified a cuvette-shaped block embedded with a very stable component that fluoresces under the same conditions as VA. This makes the calibration simple and reproducible. Simulated field tests will be used to mimic adverse conditions. Measuring Dairy
Products The ability to measure VA in dairy products will greatly increase the market for the CRAFTi. Preliminary tests indicate that the triglyceride content may interfere with VA fluorescence. We will test ionic and nonionic detergents, enzymes, and SPE to obtain an acceptable sample clean-up. Since this test would occur at the dairy processing plant, many of our concerns regarding field use are not essential for this application. They are essential, however, if this test is to be used to measure VA in breast milk of VAD women in developing countries. Testing Field Samples We have already demonstrated the CRAFTi?s utility with fortified sugar from Nicaragua. We will request samples of fortified flour from the Philippines or Africa, corn meal from Mexico, rice from Asia, and milk from the USA. In exchange for the samples, we will perform the analysis at no cost to them and provide results using the two methods. In this way, we will supplement our proof of CRAFTi accuracy. First,
these products will be analyzed by HPLC to determine sample size. Then, analysis of the same samples will be done on the CRAFTi using the sample size determined as optimal by HPLC.