The method for determing the fat content of meats must meet the following requirements to be feasible both to the packer and the inspector:

112 A S T U D Y OF METHODS O F T E S T I N G AND SAMPLING FOR THE D E T E R M I N A T I O N OF F A T C O N T E N T OF GROUND MEAT1 The determination of the fat content of ground meat is of utmost importance to the meat packers as well as the consumer The packer is desirous of producing a suitable product, but from an economic standpoint, anxious to use as much fat as possible and still be within requirements of the specifications set forth for the item, The consumer wants the ground meat component of his ration to be palatable, and not unduly wasteful The fat content is one of the most important constitutents determining this factor The method for determing the fat content of meats must meet the following requirements to be feasible both to the packer and the inspector: 1 The method of sampling must be such as to be representative of the product as a whole 2 The method of determination must be accurate 3, The results of the tests must be determined rapidly 4 Procedure must be as simple as possible The determination of the fat content of meat and meat products by ether extraction was first recorded by the Association of Official Agricultural chemists (1) in 1901 Wiley (2) states there are some fats both in animal and vegetable substances that are insoluble in ether, but they exist in minute quantities There are a l s o minute quantities of bodies not fat in foods, soluble in ether and these are included in the ether extract These solu3ility deviations may have a slight bearing on the accuracy of the ether extraction method of fat determination as set forth by the Association of Official Agricultural Chemists The method of testing for the amount of fat in ground meat by the armed forces as set forth in TB Med 233 (AFM16041) (3) has slightly modified the method of the Official Agricultural Chemists To meet the need for a rapid methdd of fat analysis, the laboratories of the meat packing plants and other personnel in laboratories interested in food inspection and research have devised several methods Oesting and mufman (4) described a rapid method used for the control of manufacturing operations Swift and Company Research Laboratories (5) devised a meehod of rapid determination of fat in meat and meat products using hydrochloric acid in distilled water for digestion of the meat 1, Contribution No 199, Department of Animal Husbandry, and No, 492, Department of Chemistry, Agricultural Experiment Station, Kansas State College

113 Armour and Company Laboratories (6) described a method used, on a t e n t a t i v e basis, as a control method in t h e i r processing operation In research work conducted by Hall, (7) a rapid method of f a t d e t e r mination of ground meat has been used wherein t h e sample was digested with g l a c i a l a c e t i c a c i d and concentrated sulphuric a c i d and centrifuged i n a Babcock b o t t l e The Army Medical Service Graduate School (8) used a method u t i l i z i n g a Paley Cheese Babcock b o t t l e, The Depot Veterinarian, Chicago Quartermaster Depot ( 9 ), were checking a method i n which t h e ground meat was digested i n a pyrex Erlenmeyer f l a s k and the f a t content measured on a 15 cc tube attached t o t h e f l a s k, without centriming METHODS AND PROCEDURES Three d i f f e r e n t groups of t e s t s were conducted The methods of obtaining and handling t h e samples used i n the t e s t s varied, i n some respects, from group t o group, A l l samples were run i n duplicate, and l a t e r rerun a s a check, Methods of t e s t i n g, a Ether e x t r a c t i o n (Method of O f f i c i a l Analysis of the Association of Agricultural Chemists) (10) b Method f o r f a t determination of ground meat without centrifuging w i n g sulphuric acid (9) &paratus shown i n Figure 1 Tengram samples were placed i n Pyrex e x t r a c t i o n Erlenmeyer f l a s k and thoroughly dispersed i n 10 m l of water (7OoC) with aid of glass s t i r r i n g rod; 5 m l concentrated a c e t i c a c i d was added and then 25 m l concentrated sulphuric The Erlenmeyer flask was gently r o t a t e d and shaken t o a i d i n digestion of t h e ground meat Complete digestion occurred i n two t o t h r e e minutes a f t e r addition of concentrated sulphuric acid Warm water (7OoC) was added t o bring the l e v e l of the contents up even with the lower edge of t h e ground g l a s s neck The measuring tube was placed i n t o p o s i t i o n and t h e surface brought t o t h e top of the s c a l e marked on t h e tube, by f u r t h e r a d d i t i o n of warm water ( 7OoC) The f a t column moved up i n t o t h e measuring tube and was measured by reading from lower l e v e l of top meniscus t o lower l e v e l of top meniscus t o lower level of bottom meniscus Figure 2 The percentage of f a t present in t h e sample was calculated by multiplyt h e number of m i l l i l i t e r s occupied by 895 ml x 895 (sp,gr) x 100 percentage lo g

314 c Rapid method of fat determination without centrifuging using Minnesota reagent (lo), which is ccmposed of, sodium salicylate, potassium cwbonate, sodium hydroxide, and isopropyl alcohol The apparatus and procedure were the same as were previously described for the new method for rapid fat determination of ground meat without centrifuging, except that 25 m l of Mnnesota reagent was used in l i e u of the acetic and sulphuric acid The flask was also heated for 15 to 20 minutes to aid in digestion of the meat d Modified Babcock method of fat determination with centrifuging using sulphuric acid (8) A 9gram sample of ground meat was placed in Paley bottle to which was added 5 m l of water (7OoC) Sample was thoroughly dispersed in water with aid of glass stirring rod and 5 m l of acetic acid was added, then 15 ml of sulphuric acid By mix ing and stirring digestion occurred in about two to three minutes After securing the stopper in the Paley bottle, it was centribottle fuged for five minutes at ap roximately 1000 rpm was place: in water bath (708C) for two minutes, and hot water (70 C) added to bring the surface of the contents up near the top of the graduation on the stem The bottle was centrifuged for two minutes at ap roximately 800 rpm, and again tempered in water bath (708C) for two minutes The percentage of fat in the sample was determined by reading from the bottom of top meniscus to bottom of lower meniscus Figure 3 e Modified Babcock method with centrifuging using Minnesota Reagent The procedure was the same as was previously described for modified Babcock method of fat determination with centrifuging except 15 ml of Minnesota reagent was used in lieu of acetic and sulphuric acid, and sample heated for 15 to 20 minutes to aid digestion Group A The samples used were prepared by using ground beef with which different amounts of fat were incorporated with thorough mixing Series of samples were prepared in duplicate; quarterpound samples were wrapged in wet wax per and frozen at O F until used for the test Methods used: a b C Ether extraction (AOAC) Rapid method without centrifuging using Minnesota reagent Modified Babcock method with centrifuging using Minnesota reagent

Group B Method of sampling was similar to the method used for Group A except some of the meat tested was ground and mixed three times, and a portion ground and mixed five times The grinding and mixing was accomplished by passing through a small hand grinder using varied sizes of plates, blending and mixing af'ter each operation Methods used: a b C Group C Ether extraction (AOAC) Rapid method without centrifuging using sulphuric acid Modified Babcock method with centrifuging using sulphuric acid All of the samples used in this group of tests were seasoned pork sausage Methods used: The ether extraction (AOAC), method and the modified Babcock method with centrifuging (using sulphuric acid), as used for the Group B series of tests OBSERVATION AND DISCUSSION In Tables 1, 2, and 3 is recorded the percent of fat found in the duplicate samples, using different methods of testing, and the results of rerun tests in 8 part of the Group B series More tests were run than shown in the tables Results for the samples excluded were not complete in duplicate and r e m tests, and comparative readings could not be made The tables and statistical analysis of results show a comparison between the standard ether extraction (AOAC) method and other methods of fat determination in ground meat, based on duplicate samples It is desired to state that a m area laboratores, when receiving duplicate samples with the ether extraction method, do not permit duplicate determlnation to exceed 5 percent in variation In fact, such variation,seldom exceeds 2 percent For the purpose of this study a wider variation in duplicate results in the ether extraction method was considered acceptable by the authors In the Group A series of samples when the Minnesota reagent was used the results ehowed the following by statistical analysis as compared with the standard ether extraction (AOAC) method 1 There was a significant difference in 100 percent of tests checked between the ether extraction (AOAC) method and method without centrifuging using Minnesota reagent 2 There was a significant difference in 80 percent of tests using modified Babcock method wlth Minnesota reagent and centrifuging, compared with the ether extraction (AOAC) method

In t h e Group B eerie6 of tests t h e correleticn betweer rerun tests within each method were dztermined ptatisticallyj shoving t h e following: 1 S i g n i f i c a n t diffeyences v e x noted between 92 percent rerun tests i n t h e e t h e r e x t r a c t i o n (AOAC) method, 2 SiSnificant differences existed b e t m e n 75 percent of rerun t e s t s using sulphuric acid, without centx9f uging, 3 Significant differences occurred betweeil r e r m tests i n 25 percent of tests with the modified Babcock method using s u Q h u r l c a c i d and centripaing 21the same grodp of t e s t s, s t a t i s t i c a l am1ysis demonstrated the following differences between methods, using a varied mixing proceduze: 1 Samples mixed f i v e times a There was a s i p i f i c a n t difference i n 91 percent of t e s t s w i n g s u l p b w i c a c i d withoct centrifuging compared with ether e x t r a c t i o n (ACCIC) method b There was a s i g n i f i c a n t dtfference i n O8 percent of t e s t s by t h e modified Dabcock rnztho8 using sulphuric acid and centrifuging compaed with the other e x t r a c t i o n (KIAC!) mzthod, 2 Sanples mixed three times a There wzs s i g n i f i c a n t difference i n 37 percent of tests using s u l t h r i c a c l 3 w i t h m t aentrifuging compared with t h e ether e x t r a c t i o n (AOAC) method 7 + b Taere m s s i g n i f i c a n t difference i n 75 percent of tests by the modified Babcock method, using sul h u r i c a c i d and centrifuging compared w5th t h e e t h e r exkrctctlor (AOAC) metho In G r m q C s e r i e s of tests all flainvles were pozk sausage which had beea sz&soaed, which making t h e reading of t h e f a t column in the Paley t e s t tube d i f f i c u l t t o determine because t h e seasoning c o l l e o t s a t the bottom of t h e f a t i n t h e tube, By s t a t i s t i c a l a n a l y s i s 54 percent of +he tests showed r;5p:',ficsrr: differences betwwn *,ha e t h e r e x t r a c t i o n (AOAC) method and t h e modili ed Baboock me5hod u s i r g sulphvric a c i d and centrifuging S W M AND CONCLUSION I _ On t h e b?s,sis of t h e findings of results, i n the tests conducted, t i e rollowing c o x l u s i o n s ca~zbe mzde: 1, Thers was less s i g n i f i c a n t difference between the r e p e t i t i o n tests using t h e Modified Babcock method of reipid fat determination; with centrifu&in&the samples G%er digestion with sulphuric a c i d than when the e t h e r e x t r a c t i o n (AOAC) method m s used The o v e r a l l rssults obtained by t h e two methods compared closs2y when t h e saaples were w z l l mixed 2 * The rapid methoc? of f e t determination without centrifuging a f t e r dtgestioc with sillphuric a c i d was not e s a t i s f ' a d o q method of fat

117 determination in ground meat The results vary from results obtained by the ether extraction (AOAC) method; a l s o there is a wide variation between correlation tests 3 The use of the Minnesota reagent in the Modified Babcock method with centrifuging and the rapid method of fat determination without centri fuging is not satisfactory because of variable results and inability to bring about complete disintegration of the tissues and release of all the fats 4 Complete mixing and grinding of the samples is essential in order to get as representative a sample as possible for testing, regardless of the method employed 5 Seasoning added to pork sausage interferes with the accuracy of the fat determination by the Modified Babcock method The lower meniscus of the fat column in the neck of the Paley test tube is not distinct for reading 1 Bureau of Chemistry Food analysis United States Department of Agriculture Bulletin 65, Washington, D*Cj 1902 2 3 4 5 Wiley, Harvey W Foods and their adulteration, Philadelphia: P a Blakiston's Son and Company, 1907 Department of the A r q r Technical Bulletin TB Med 233 (Department of the Veterinary Laboratory service (food analysis) Department of the Army and Air Force, 1950 Alr Force Manual APM 16041) Oesting, R B and 1, P Kaufman Rapid determination of fat in meat and meat products Analytical edition, Ind Eng Chem 17, 125, 1945 Swift and Company Research Laboratories Fat nest and Meat Products, Using Babcock Bottles Method Ca4C49 unpublished Obtained by communication with VC Mehlenbacker, Chemist, Swift and Company, Union Stock Yards, Chicago, Illinois, January, 1952 6 Armour and Company Laboratories Rapid Control Method for the Determination of fat in Cooked and Uncooked Sausage and Ground Meat such as Hamburger (Tentative) Unpublished Obtained by communication with L A Michael Armour and Company Februab 6, 1951 Laboratory; Armour and Company, Kansas City,'Kansas, 7 Hall, J LOW(? Babcock Method for Determining fat in Meat Products Unpublished Obtained by communication with J Lowe Hall, Meat Research, Chemistry Department, Kansas State College, Manhattan, Kansas, October, 1951 8 Army Medical Service Graduate School, Arqy Medical Center Rapid Method for fat determination in Ground Meat Unpublished Obtained from Director, Veterinary Division, Army Medical Service Graduate School, Army Medical Center, Washington, DC, October, 1951

9 Depot Veterinarian Office, Chicago Quartermaster Depot New Method for Fat Determination of Ground Meat Unpublished Obtained from Depot Veterinarian, Chicago Quartemgter Depot, Chicago, Illinois, November, 1951 10 Association of Official Agricultural Chemists Methods of Analysis Washington: Association of Official Agricultural Chemists, 1950

119e Table 1 Results of Tests Conducted on Group A Series of Samples SEMple Series Ether Extraction W/O Centrifuging Modified Babcock using (AOAC) using Minnesota Minnesota Reagent 1 1 3735 35e35 35 23 35 71 27 e15 4 4 27 32 3501 3375 3370 2 2 3 3 5 5 26e4S 32 e 22 2000 20e 00 2700 27e50 2700 2700 31 50 3150 3222 20 rn 58 2058 1967 20e 58 2685 2774 20 58 20 58 22 e o 0 22 00

120 Table 2 Result6 of Tests Condused on Group B Series of Samples I _ Sample Series Ether Ekbraction (AWC) 1A 1B W/O Centrifuging using sulphuric acid 2A 2B Modified Babcock using suipbur3c acid 3A 3B Samples Ground and Mixed Five Times 6 6 7 7 8 8 9 9 10 10 11 11 12 12 13 13 14 14 15 15 16 16 17 17 18 18 19 19 20 20 21 21 22 22 23 23 24 24 25 25 25 20 24,90 3410 3430 3680 3680 27 a 1 0 2690 3896 3896 38 96 39 97 4310 4310 4460 4570 4590 4730 4060 4130 4870 49,50 3820 3910 2540 2237 25 lo 2327 3450 33 e 1 1 34,50 33 11 3660 33 ll 3660 3132 25 a 0 0 2550 25 50 25 50 4260 3301 3893 3890 3850 4220 3860 3850 3301 3893 3490 38 90 3990 37 a59 4530 4300 4400 4117 4206 4530 4300 4350 4161 4206 4940 4550 5100 4475 4877 4800 4400 5100 4385 4778 4920 4206 5057 4710 4750 5110 4520 4750 4251 5101 4060 3980 4l00 3803 4161 4080 4050 4000 41e17 4161 4630 4750 4500 4475 5101 4670 4206 4117 4700 4950 4320 4100 4150 3893 3759 4330 4100 4060 3803 3759 Samples Ground and Mixed Three Times 2990 2550 27 74 30 20 2580 2775 4050 4510 4400 4340 4117 4296 4050 4510 4257 4300 4117 4250 3820 4300 3840 3950 3848 3938 3740 43000 3840 4040 3848 3938 46054 49040 4800 4370 4470 42 51 4660 4800 4400 4470 4200 4280 4117 4250 3840 4800 3840 4800 4217 4270 4251 4117 3910 4350 3770 4160 3803 4206 3910 4350 3770 4160 3048 4206 4740 4800 4654 4654 3700 4620 4740 4850 4698 5056 3700 4620 3900 4050 3790 4160 3938 3893 3950 3950 4027 3893 3950 4230 I 0 0 L

121 Table 3 Results of Test Conducted on Group C Series of Samples Sample Series 26 26 27 27 28 28 29 29 30 30 31 31 32 32 33 33 34 34 35 35 36 36 Ether Extraction Modified Babcock (AQAC) Using Sulphuric Acid Percent f a t 3170 31m 60 3080 30 90 32lo 32010 2650 2680 35 oo 35 m00 45 080 4590 39 60 3930 37 90 38e 0 0 47080 4790 30 30 3030 28 70 29 010 Percent f a t 3110 3000 3450 3350 32 40 3140 3140 33 50 3490 3500 43e 1 0 43 90 39 e80 38010 3650 3650 3610 3600 2950 3950 29 50 2950 MQDEBA!i GR ~ I C K S O N ; The next p a r t of t h e programt wh,~:h is on e t h e r extraction, w i l l be given by my good f r i e n d from Kansas State College, Dr Hall #######