MICROTRACERS F - Testing for "Cross Contamination" in Medicated Feeds.

Please also refer to Literature Items "L" Quality Assurance With Microtracers F,
 "N" Microtracer "Rotary Detector", "O" Microtracers F- Quantitative Assays" and 
"MM" The Use of Microtracers F in 
Coding the Presence of Coccidiostats in Poultry Feeds: Practical Implications. .

Preface: Medicated animal, poultry and aquatic feeds are often manufactured at
feedmills making feeds for many species or if not at least many formulations for
 one species. Contamination of medications to non-target feeds is inevitable 
but quantifiable and controllable. Medicated feed assays, unfortunately, often 
do not offer a viable control mechanism because: they are expensive, they 
cannot be performed immediately and they are often inaccurate at 
contamination levels (i.e. 1 % or less of formulated levels).

Microtracers F (colored uniformly sized iron particles) offer a vehicle for locating
 and quantifying "cross contamination" of medicated feeds. Instead of running expensive
 and inaccurate drug assays, one uses one or more Microtracers F as indicators for the
coded medication. The tracer is first mixed into the medicated feed premix, then the premix 
is added to the feed and mixed. Many samples are taken either only at truck loading or at
 many locations, depending upon the scope of the study. It is critical that feeds 
not formulated with the Microtracer follow the identical route at the feedmill as the 
"target" batch containing the Microtracer. Samples yielding significant levels of the
 tracer may then be analyzed for the medication to confirm (or refute) the
 preliminary results provided by the tracer.

Addition of the Tracer: The Microtracer is generally added to the medicated 
premix to yield 50 grams of tracer per metric tonne of final feed. Since the
 tracer has an expected count of 25,000 particles/gram, this means 
50 X 25,000 or 1,250,000 colored iron particles are added to a one metric ton
 batch with 1,250 particles expected per kilogram of feed or 125 per 100 grams, 
assuming the feed is completely mixed, the tracer is stable and it meets its specified count.

Sampling: One should take a minimum of four and preferably ten or more "grab" 
samples from the target batch formulated with the Microtracer. Samples of the
 target batch should weigh at least 200 grams and ideally be taken at several points
 in the feed production system. If one analyzes 80 gram "grab" sub-samples of each
 target sample, one will expect to find 100 tracer particles per sample.

One should then take samples from one or more following batches not formulated with the
 medication or the tracer. These samples should weigh at least 1-kilo each. If one analyzes
800 grams of a feed supposed to contain no tracer and finds 100 tracer particles, then if
 total tracer recovery is 100% of theory one may estimate 10% of the total tracer 
(and thereby the drug) was found as contamination in that sample. By analyzing
 very large samples of feeds supposed to contain no tracer, the "sensitivity" 
(ability of the tracer to allow accurate detection of very low level contamination) 
may be improved by a factor of 10 or more. Tracer contamination at 0.01 is possible!

Page 2 Microtracers F- Testing for "Cross Contamination" of Medicated Feeds

Individual batches of feed must be isolated and their flow controlled. People must be
 ready at each sampling location to take samples as the individual batches pass. 
All samples must be carefully identified and detailed records of all aspects of the test should be made.

The basic flow of feed at a feedmill includes the following:

1. feed ingredients after being ground to mash with a hammermill (if necessary)
 are gravity fed into a mixer and mixed.
 2. the mixed feed is then dumped into a surge bin.
 3. the feed is then discharged from the surge bin using a screw conveyer.
4. the feed is then elevated in the feedmill using a bucket elevator system.
5. the feed is then gravity dumped into a holding bin over a pellet mill.
6. the feed is then pelleted and cooled.
7. the feed is then elevated again using a bucket elevator or pneumatic system.
8. the feed is then distributed to holding bins. 
9. the feed is then loaded into trucks. 
10. the feed is then carried to farms where it is discharged into bins. 
11. the feed is then distributed from the on-farm bins to the animals, poultry or fish for consumption.

Medications are usually added as a microingredient into the mixer, sometimes by hand 
but often utilizing computer controlled micro-bin systems. "Cross Contamination" 
of the medication may occur in the micro-bin system even before the medication reaches the mixer.

"Cross contamination" to prior batches may also occur, as when mixer discharge gates leak 
and an earlier batch remains in the surge bin.

As an initial investigation, one may take samples of the "non-target" following feed
 only at truck loadout taking care to be certain the feed takes the identical route
 as the medicated feed with Microtracer. If one finds very little or no tracer from
 the loadout samples, this indicates there may be very little "cross contamination"
occurring anywhere in the system. If one finds Microtracers in the "non-target" 
following feed, however, much more extensive testing is required to diagnose and 
then have a basis for correcting the problem.

Portions of the medication may be left as "contamination" to non-target batches 
at all the above referenced production points. To reliably diagnose the potential for
 "cross contamination" of medications to non-target batches of feed, it is critical to
 take multiple "grab" samples at multiple locations of both the target batch
(with medication) as well as one or more following batches. It must be re-emphasized
 that it is critical the following batches take the identical route through the feedmill
 as the initial medicated batch formulated with Microtracer.


Page 3 Microtracers F - Testing for "Cross Contamination" of Medicated Feeds

Analysis of Samples: Please refer to Microtracer literature items "L" and "O".

Results from Actual Tests

Test # 1. Premix Plant manufacturing Powdered Sulafethazine premix. Microtracer  
F-Red was used to estimate contamination of powdered sulfamethazine to flushing materials
 and to subsequent premix batches. The premix plant had been "red flagged" by the
 US-FDA because it found more than 5 ppm sulfamethazine in premix supposed to contain none
 The premix plant had instituted rigorous flushing cleanout procedures 
 and wanted to confirm these were efficacious.


Premix Formulation- 1 % sulfamethazine (10,000 ppm) and Microtracer F-Red at 0.8% (8,000 ppm).
 Batch size 2,500-lbs.

Batch #1-                            Microtracer F-Red Counts                    Sulfamethazine Assay
Sample  #1- 0.41 grams =                  66
              #2- 0.40   "                          73
              #3- 0.47   "                          64
              #4- 0.38   "                          56
Total 1.66 grams =                           259                             Composite Assay 8,800 ppm
Percentage of Specification              77%                                             88%
(25,000 X 1.66 grams X 0.8% = 332 expected count - 259/332 = 77%)

Flush #1. 300-lbs. of ground limestone.
Sample #1- 20.0 grams =                 85
            #2- 
        "                                        86
            #3-      
"                                      104
            #4-       "         
                           88
Total            80.0 grams =              363                               Composite Assay  331 ppm
Percentage of Specification            2.3%                                          3.3%
(25,000 X 80 X 0.8% = 16,000 expected count - 363/16,000= 2.3%)

Flush #2. 300-lbs. of ground limestone.
Sample  #1- 200 grams =             106
             #2-       "                           90
             #3-      
"                                 109
             #4-       "                         101
Total            800 grams =             406                             Composite Assay   82 ppm
Percentage of Specification         0.25%                                     0.82%


Page 4 Microtracers F - Determining "Cross Contamination" of Medicated Feeds

Flush #3. 300-lbs. of ground limestone.
                                      Microtracer F-Red Counts                  Sulfamethazine Assay
Sample  #1          200 grams = 16
            #2- 
             ''                    30
            #3-              "              14
           #4-              
"                   27
Total                   800 grams    87                                      Composite Assay  65 ppm
Percentage of Specification  0.054%                                               0.65%

Flush #4. 1,000-lbs. of ground corn.
Sample #1-        200 grams     28
            #2-           "
                       22
            #3-           "           
        22
            #4-           " 
                     23
Total                800 grams       95                                     Composite Assay     32 ppm
Percentage of Specification 0.059%                                                0.32%

Flush #5. 1,000-lbs. of ground corn.
Sample # 1-     200 grams     2
            #2-            "
                   2
            #3-            "    
              5
            #4-          
"                  11
Total             800 grams       20                                        Composite Assay      10.6 ppm
Percentage of Specification 0.0125%                                        0.106%

Followup Batch #1. 2,500-lbs.
Sample #1- 200 grams      4
            #2-    " 
                       2
            #3-    "                  1
           #4-     "                  2
Total           800 grams      9                                              Composite Assay     1.7 ppm
Percentage of Specification 0.0056%                                                       0.017%


Page 5 Microtracers F- Determining "Cross Contamination" of Medicated Feeds

Conclusion: The various "flushes" did serve a useful purpose in reducing the
 amount of tracer (and sulfamethazine) reaching premixes supposed to contain none
 Contamination was not uniform but rather erratic when present at very low levels. 
It appeared possible to keep sulfamethazine contamination 
of following premixes below the objective of 5 ppm.

"Cross Contamination" of the Microtracer F-Red from the premix 
formulated with the tracer to subsequent flushes and premix batches
was less than for the powdered sulfamethazine. It should be possible to make 
a curvilinear regression to accurately estimate contamination of the drug 
based upon tracer results. The results also may support the advantage 
of granulated sulfamethazine over the powdered product 
as relates to their potential for cross contamination.


Total recovery of the Microtracer F-Red was about 80% of specification
and total recovery of the sulfamethazine was about 90%. Most probably, 
the Microtracer was lower than its specified count.The test continued through
 four more production batches of premix. Microtracer was found in all 
batches and sulfamethazine assays of the final batches was 2.4ppm
 and 2.5ppm (0.024% and 0.025% of the specified level).

Test #2. Using Microtracers F to Design a Feedmill Identifying Locations 
Where the Greatest Contamination Occurs. A major designer/supplier 
of "turnkey" feedmills wanted to minimize batch to batch contamination in their new mills.

Microtracer F-Blue was formulated in "Batch #1" at 50 grams per 2,000-lbs. 
of feed into 3 ton batch (150 grams of tracer mixed in ground corn added to batch).
 Microtracer F-Red was formulated following the same procedures into Batch #2
 Samples were then taken at the Surge Bin, Conveyer and Top of the Bucket Elevator
 for five following batches.


Microtracer F-Blue Contamination to Subsequent Batches.

Batch (2,000 grams)          Surge Bin            Conveyer             Elevator
         #2- sample 1                  2                         1                      17
                           2                  1                         0                      98
                           3                  1                         3                    112
                           4                  0                         3                     58
Total                                       4                         7                    285
Percentage of Tracer Spec. 0.035%              0.065%              2.59%


Page 6 Microtracers F- Determining "Cross Contamination" of Medicated Feeds

Batch                                Surge Bin              Conveyer               Elevator
#3-       sample 1                    0                          3                          10
                        2                    2                          0                            9
                        3                    0                          0                           23
                        4                    0                          1                           44
Total                                      2                          4                           86
Percentage of Tracer Spec. 0.018%             0.035%                    0.78%

#4-         sample 1                 2                           0                            2
                          2                 0                           0                            0
                          3                 0                           0                            0
                          4                 0                           0                          15
Total                                     2                           0                          17
Percentage of Tracer Spec. 0.018%                  0%                     0.155%

#5- sample 1                        0                            0                            0
                  2                        1                            0                            0
                  3                        0                            0                            1
                  4                        0                           1                             5
Total                                    1                           1                             6
Percentage of Tracer Spec. 0.009%             0.009%                 0.055%

                                         Surge Bin              Conveyer              Elevator
#6- sample 1                            0                        0                         1
                  2                            0                        0                         1
                  3                            0                        0                         1
                  4                            0                        2                         6
Total                                        0                        2                         9
Percentage of Tracer Spec.      0%              0.018%              0.082%

#7- sample  1                            1                       1                        0
                   2                            1                       0                        0
                   3                            0                       0                     129
                  4                             0                       0                         0
Total                                          2                      1                     129
Percentage of Tracer Spec. 0.018%            0.009%               1.17%

Conclusion: Comparatively little contamination of the Microtracer occurred 
at the Surge Bin and Conveyer locations. The amount of contamination was
 much greater at the top of the bucket Elevator. One sample in Batch #7
 contained a "slug" of tracer illustrating that contamination is
not uniform and thee best chance to find problems is to take and 
analyze many large samples. The amount of contamination increases
 as the feed flows through the mill.


Page 7 Microtracers F- Determining "Cross Contamination" of Medicated Feeds

In detailed studies at three poultry feed mills (ref. Item "MM"), the following results were obtained:

Recovery of Microtracer F-Red from samples taken from the mixers - 95.7% of specified count

Recovery of Microtracer from pelleted feed samples at truck loading- Feedmill # 1- formula one 82.8%, 
formula two- 89.3%; Feedmill #2- 75.4%.

Percentage of tracer appearing in non-target feeds- Feedmill # 1- 3.0%, Feedmill #2- 0.26%. 
The medicated premix at Feedmill # 1 was formulated in 60% of the mills feeds whereas the
 medication at feedmill #2 was formulated in only 10% of its production.

Qualitatively, the Microtracer results reflected feed medicated assays well,
 though quantitatively results did not match as well.
This was because of variability inherent in the particle statistics used to interpret
 Microtracer spot counts and problems with the medicated feed assays.

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