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Total Folate, Pantothenic Acid and Biotin in Some Fish Products
Can. Inst. FaadSci. Technal. J. Vol. 22, No. 2, pp. 170-172, 1989
RESEARCH
Total Folate, Pantothenic Acid and Biotin in Some Fish Products K. Hoppner and B. Lampi Nutrition Research Division Food Directorate Health Protection Branch Health and Welfare Canada Ottawa, Ontario, Canada KlA OL2
Health Protection Branch. As part of the protocol, samples were divided into summer and winter periods each year from different cities across Canada. The present samples were obtained from collections made during 1985 in Ottawa, 1986 in Halifax and 1987 in Winnipeg.
Abstract Total folate, pantothenic acid and biotin levels were determined in different raw and processed fish products by microbiological assay. The data for these vitamins in the raw and processed fish products provide additional information for improving the data base of the Canadian Nutrient file.
Representative homogenized samples were obtained from a central kitchen where they were prepared according to normal household practices. Non-shellfish samples were analyzed raw, cooked or baked. Shellfish samples were analyzed raw or cooked, and smoked and canned samples as received. All samples were homogenized, weighed and freeze-dried for moisture determination and vitamin analysis.
Resume Les concentrations de folacine IOtale, d'acide pantothenique et de biotine ont ete determinees dans divers poissons crus ainsi que leurs derives, par la methode microbiologique. Ces nouveaux taux de vitamines serviront a ameliorer la base de donnees du Fichier canadien sur les elements nutritifs.
Introduction
For total folate analysis, duplicate extracts for deconjugation were prepared as previously described (Hoppner and Lampi, 1982). Total folate activity was determined by microbiological assay with L. casei ATCC 7469 and "Difco" assay medium (Difco Laboratories, Detroit, MI) with added ascorbate (0.25010 (w/v) final assay media) (Baker and Frank, 1967; Herbert and Bertino, 1967). The assay tubes were incubated for 22 h at 37°C and measured turbidimetrically at 660 nm.
The metabolic functions and human nutritional requirements for folate, pantothenic acid and biotin for maintainance of good health have recently been reviewed (Department of National Health and Welfare, 1983; Fox, 1984; Bonjour, 1985). Evaluating diets for folate, pantothenic acid and biotin has always been very difficult because of lack of current and complete information available on the composition of these vitamins in raw and processed foods. Since its development in 1979 the Canadian Nutrient File has served to compile and provide up-to-date nutrient data for Canadian users (Verdier and Bear-Rogers, 1984). In order to add to the current data base, this paper reports the folate, pantothenic acid and biotin content in raw and processed fish products obtained from the collection of foods for a total diet study underway in the Food Research Division, Health Protection Branch.
Total pantothenic acid was determined microbiologically with L. plantarum ATCC 8014 using Difco A.O.A.C.- V.S.P. assay medium (Zook et al., 1956). For the release of bound pantothenic acid, duplicate extracts were prepared with the double enzyme system of 2% intestinal phosphatase and 10% Dowex-1X8 treated pigeon liver extract (Sigma, St. Louis, MO) (Wyse et ai, 1985). The weight of d-caIcium pantothenate was adjusted in terms of pantothenic acid for the preparation of the standard solutions. Incubation was carried out at 37°C for 22 h and the turbidity measured at 660 nm.
Materials and Methods The samples of the fish products were obtained from a total diet study presently underway in the
Copyright Cl 1989 Canadian Institute of Food Science and Technology
170
Table I. Folacin activity in raw and processed fish products. Sample Fish Product (n) Cod Haddock Halibut Lobster Pickerel Salmon Sardines Scallops Shrimp Trout
Tuna Whitefish
-raw/frozen -baked -smoked -raw/frozen -baked -raw/frozen -raw/frozen -cooked -raw/frozen -baked -raw/frozen -canned -canned -raw/frozen -cooked -raw/frozen -cooked -raw/frozen -baked -smoked -baked -canned -raw/frozen -baked
Moisture Ufo Mean + S.D.
8 4
76.8 68.2 73.4 78.8 75.2 75.6 77.8 76.7 79.0 74.6 66.1 68.0 62.3 75.3 72.5 84.3 79.2 69.8 73.4 67.5 67.9 69.0 75.2 76.6
I
6 I
3 2 I
4 I
3 5 5 3 I 6 4 3 2 2 2 5 2 2
Biotin was assayed microbiologically with L. plantarum ATCC 8014 and Difco-Biotin assay medium (Skeggs, 1963). Acid hydrolysis was carried out on duplicate samples with 2N H 2S04 at 121°C for 2 h. The suspensions were adjusted to pH 4.5 with 2N NaOH, diluted to volume and filtered (Whatman No.40). Further dilutions for assay were made as required. The assay tubes were incubated at 37°C for 22 h and the turbidity measured at 660 nm.
Results and Discussion The values for total folate, pantothenic acid and biotin as determined by microbiological assay in the
Folacin activity, ug/IOOg Range 5.2-10.2 10.1-13.3
Mean ± S.D.
± 5.2 ± 9.8
± ±
1.8 1.5
± 2.6
6.1-12.0
±
2.0
± 2.9 ± 8.6
7.6- 9.8 8.1-24.2
± 2.3
2.7- 8.5
± ± ± ±
1.0 0.7 2.8 6.7
2.4- 5.0 9.0-15.7 3.9-13.0 7.4-15.7
± ± ± ± ± ± ± ± ±
4.6 4.6 6.9
3.1-12.8 5.9-10.7 5.1-12.4 4.8-15.7 9.3-10.9 11.8-12.7 2.7- 6.6 2.4- 3.6 3.3- 5.3
I.I
0.2 0.6 2.2 2.2 3.7
8.2 11.1 2.9 8.9 8.0 8.6 16.1 9.0 6.3 6.6 3.4 13.6 7.7 10.7 17.8 8.0 8.5 8.3 10.2 10.1 12.2 4.6 3.0 4.3
± 1.1 ± 11.3 ±
3.2
± ± ± ±
1.4 2.8 3.8 4.4
± ± ± ± ± ± ± ± ±
4.2 2.3 3.7 7.8 1.1 0.6 1.7 0.9 1.4
different raw and processed fish products are shown on a fresh weight basis in Tables 1, 2 and 3, respectively. In general, where comparison with published levels of these vitamins in fish products was possible, the values compared favourably (Orr, 1969; Perloff and Butrum, 1977; Paul and Southgate, 1978). Although this study was not designed to measure vitamin losses due to preparation in the same sample of the different fish products, tentative estimates of lower cooking losses of folate, pantothenic acid and biotin have been reported for fish than for vegetables and meat (Paul and Southgate, 1978). In the case of canned samples
Table 2. Pantothenic acid in raw and processed fish products. Fish Product
Sample (n)
Cod
Haddock Halibut Lobster Pickerel Salmon Sardines Scallops Shrimp Trout Tuna Whitefish
-raw/frozen -baked -smoked -raw/frozen -baked -raw/frozen -raw/frozen -cooked -raw/frozen -raw/frozen -canned -canned -raw/frozen -cooked -raw/frozen -cooked -raw/frozen -smoked -baked -canned -raw/frozen -baked
Can. Insl. Food Sei. Teehnol. J. Vol. 22, No. 2, 1989
7 4 I 5 I 2 I I
4 2 4 4 I I
4 3 3 2 2 4 2 I
Moisture 010 Mean ± 5.0. 77.0 ± 5.5 68.2 ± 9.8 73.4 79.2 ± 2.6 75.2 76.0 ± 3.9 83.8 76.7 79.0 ± 2.3 65.7 ± I.I 67.8 ± 1.2 6.63 ± 3.1 82.9 72.5 85.5 ± 3.2 77.1 ± 2.1 69.8 ± 6.9 67.5 ± 0.2 67.9 ± 0.6 68.7 ± 2.4 75.2 ± 2.2 74.0
Pantothenic acid, ug/lOOg Range 189- 334 198- 276
Mean ± 5.0. ± ±
50 40
178- 251
±
30
±
10
297- 314 239- 628 977-1064 606- 828 432- 640 34- 106 82- 142 968-2326 1339-1346 1213-1642 167- 282 866-1209
256 223 227 221 247 305 2398 1041 356 1021 744 511 149 161 80 113 1724 1342 1427 218 1037 891
± 180 ± 60
± 100 ± 90 ± 30 ± 30 ± 690 ± 10 ± 300 ± 50 ± 240
Hoppner and Lampi / 171
Table 3. Biotin in raw and processed fish products. Fish Product
Cod Haddock Halibut Lobster Pickerel Salmon Sardines Scallops Shrimp Trout Tuna Whitefish
-raw/frozen -baked -smoked -raw/frozen -baked -raw/frozen -raw/frozen -cooked -raw/frozen -raw/frozen -canned -canned -raw/frozen -cooked -raw/frozen -cooked -raw/frozen -smoked -baked -canned -raw/frozen -baked
Sample (n) 7 4 I
5 1 2 1 1 4 2 4 4 1 1 4 4 3 2 2 4 2 1
the addition of vitamins through other ingredients could distort such estimates. The profile of folate derivatives and polyglutamate chain length has been reported for a few selected foods but not for fish (Rodriguez, 1978). It is presently not known whether some of the differences in vitamin content as indicated for pantothenic acid is species related, due to feeding habit or possibly commercial rearing. Data for the vitamin content of fish are scarce and not all foods have been analyzed for the above vitamins, particularly biotin. It is hoped that many of the gaps in data for these vitamins can be filled and thereby improve the data base of the Canadian Nutrient File.
Acknowledgements The authors thank the staff of the Total Diet Study for their technical assistance in the collection and preparation of the samples. References Baker, H. and Frank, O. 1967. A microbiological assay for folate activity. In: Vitamins, Chemistry, Physiology, Pathology, Methods. Vol. VII, 2nd ed. P. Gyorgy and W.N. Pearson (Eds.) p. 269. Academic Press, NY. Bonjour, J.P. 1985. Biotin in human nutrition. Ann. N. Y. Acad. Sci.447:97. Department of National Health and Welfare. 1983. Recommended Nutrient Intakes for Canadians. Bureau of Nutritional Sciences, Food Directorate, Health Protection Branch, Ottawa, Ont.
172 / Hoppner and Lampi
Moisture 070 Mean ± S.D. 77.0 ± 5.5 68.2 ± 9.8 73.4 79.2 ± 2.6 75.2 76.0 ± 3.9 83.8 76.7 79.0 ± 2.3 65.7 ± 1.0 67.8 ± 1.2 62.6 ± 3.1 82.9 72.5 86.7 ± 1.8 79.2 ± 4.6 69.8 ± 6.9 67.5 ± 0.2 67.9 ± 0.6 68.7 ± 2.4 75.2 ± 2.2 74.0
Biotin, ug/100g Range 1.1- 2.8 2.0- 3.2
2.0- 3.4 3.0-3.3 1.4- 2.5 6.7- 8.1 7.7-12.5 8.4-10.1 0.3- 0.7 0.5- 1.0 3.3- 6.5 4.0- 4.3 2.6- 3.6 1.1- 3.3 2.7-5.9
Mean ± S.D. 2.2 ± 0.6 2.6 ± 0.5 1.1 2.5 ± 0.5 2.5 3.1 ± 0.2 4.5 6.8 1.9 ± 0.6 7.4 ± 1.0 9.4 ± 2.1 9.1 ± 0.8 1.1 1.5 0.5 ± 0.2 0.8 ± 0.2 4.5 ± 1.7 4.1 ± 0.2 3.1 ± 0.7 2.1 ± 0.9 4.3 ± 2.3 4.2
Fox, H.M. 1984. Pantothenic acid. In: Food Science and Technol· ogy, Hand Book of Vitamins: Nutritional, Biochemical and Clinical Aspects. L.J. Machlin (Ed.). pA37 Marcel Dekker Inc., New York, NY. Herbert, V. and Bertino, J .R. 1967. In: Vitamins, Chemistry, Physiology, Pathology, Methods. Vol. VII, 2nd ed. P. Gyorgy and W.N. Pearson (Eds.). p. 243. Academic Press, New York, NY. Hoppner, K. and Lampi, B. 1982. Total folacin activity in breakfast cereals. Nutr. Rep. Int. 26:495. Orr, M.L. 1969. Pantothenic acid, vitamin B6 and vitamin B12 in foods. Home Econ. Res. Rep. 36, U.S.D.A., Washington, D.e. Paul, A.A. and Southgate, D.A.T. 1978. The Composition of Foods. R.A. McCance and E. Widdowson (Eds.). Her Majesty's Stationary Office, London, UK. Perloff, B.P. and Butrum, R.R. 1977. Folacin in selected foods. J. Am. Dietet. Assoc. 70: 161. Rodriguez, M.S. 1978. A conspectus of research of folacin requirements of man. J. Nutr. 108:1983. Skeggs, H.R. 1963. Biotin. In: Analytical Microbiology. F. Kavanagh (Ed.). pA21. Academic Press. New York, NY. Verdier, P. and Beare-Rogers, J.L. 1984. The Canadian Nutrient File. J. Can. Dietet. Assoc. 45:52. Wyse, B.W., Song, W.O., Walsh, J.H. and Hansen, R.G. 1985. Pantothenic acid. In: Methods of Vitamin Assay. J. Uugustin, B.P. Klein, D. Becker and B.P. Venugopal (Eds.). p. 399. John Wiley and Sons, New York, NY. Zook, E.G., MacArthur, J.M. and Toepfer, E.W. 1956. Pantothenic acid in foods. U.S.D.A. Agric. Handbook 97.
Submitted October 24, 1988 Revised December 20, 1988 Accepted January 2,1989
J. Inst. Can. Sci. Technol. Aliment. VD!. 22. No. 2. 1989
RESEARCH
Total Folate, Pantothenic Acid and Biotin in Some Fish Products K. Hoppner and B. Lampi Nutrition Research Division Food Directorate Health Protection Branch Health and Welfare Canada Ottawa, Ontario, Canada KlA OL2
Health Protection Branch. As part of the protocol, samples were divided into summer and winter periods each year from different cities across Canada. The present samples were obtained from collections made during 1985 in Ottawa, 1986 in Halifax and 1987 in Winnipeg.
Abstract Total folate, pantothenic acid and biotin levels were determined in different raw and processed fish products by microbiological assay. The data for these vitamins in the raw and processed fish products provide additional information for improving the data base of the Canadian Nutrient file.
Representative homogenized samples were obtained from a central kitchen where they were prepared according to normal household practices. Non-shellfish samples were analyzed raw, cooked or baked. Shellfish samples were analyzed raw or cooked, and smoked and canned samples as received. All samples were homogenized, weighed and freeze-dried for moisture determination and vitamin analysis.
Resume Les concentrations de folacine IOtale, d'acide pantothenique et de biotine ont ete determinees dans divers poissons crus ainsi que leurs derives, par la methode microbiologique. Ces nouveaux taux de vitamines serviront a ameliorer la base de donnees du Fichier canadien sur les elements nutritifs.
Introduction
For total folate analysis, duplicate extracts for deconjugation were prepared as previously described (Hoppner and Lampi, 1982). Total folate activity was determined by microbiological assay with L. casei ATCC 7469 and "Difco" assay medium (Difco Laboratories, Detroit, MI) with added ascorbate (0.25010 (w/v) final assay media) (Baker and Frank, 1967; Herbert and Bertino, 1967). The assay tubes were incubated for 22 h at 37°C and measured turbidimetrically at 660 nm.
The metabolic functions and human nutritional requirements for folate, pantothenic acid and biotin for maintainance of good health have recently been reviewed (Department of National Health and Welfare, 1983; Fox, 1984; Bonjour, 1985). Evaluating diets for folate, pantothenic acid and biotin has always been very difficult because of lack of current and complete information available on the composition of these vitamins in raw and processed foods. Since its development in 1979 the Canadian Nutrient File has served to compile and provide up-to-date nutrient data for Canadian users (Verdier and Bear-Rogers, 1984). In order to add to the current data base, this paper reports the folate, pantothenic acid and biotin content in raw and processed fish products obtained from the collection of foods for a total diet study underway in the Food Research Division, Health Protection Branch.
Total pantothenic acid was determined microbiologically with L. plantarum ATCC 8014 using Difco A.O.A.C.- V.S.P. assay medium (Zook et al., 1956). For the release of bound pantothenic acid, duplicate extracts were prepared with the double enzyme system of 2% intestinal phosphatase and 10% Dowex-1X8 treated pigeon liver extract (Sigma, St. Louis, MO) (Wyse et ai, 1985). The weight of d-caIcium pantothenate was adjusted in terms of pantothenic acid for the preparation of the standard solutions. Incubation was carried out at 37°C for 22 h and the turbidity measured at 660 nm.
Materials and Methods The samples of the fish products were obtained from a total diet study presently underway in the
Copyright Cl 1989 Canadian Institute of Food Science and Technology
170
Table I. Folacin activity in raw and processed fish products. Sample Fish Product (n) Cod Haddock Halibut Lobster Pickerel Salmon Sardines Scallops Shrimp Trout
Tuna Whitefish
-raw/frozen -baked -smoked -raw/frozen -baked -raw/frozen -raw/frozen -cooked -raw/frozen -baked -raw/frozen -canned -canned -raw/frozen -cooked -raw/frozen -cooked -raw/frozen -baked -smoked -baked -canned -raw/frozen -baked
Moisture Ufo Mean + S.D.
8 4
76.8 68.2 73.4 78.8 75.2 75.6 77.8 76.7 79.0 74.6 66.1 68.0 62.3 75.3 72.5 84.3 79.2 69.8 73.4 67.5 67.9 69.0 75.2 76.6
I
6 I
3 2 I
4 I
3 5 5 3 I 6 4 3 2 2 2 5 2 2
Biotin was assayed microbiologically with L. plantarum ATCC 8014 and Difco-Biotin assay medium (Skeggs, 1963). Acid hydrolysis was carried out on duplicate samples with 2N H 2S04 at 121°C for 2 h. The suspensions were adjusted to pH 4.5 with 2N NaOH, diluted to volume and filtered (Whatman No.40). Further dilutions for assay were made as required. The assay tubes were incubated at 37°C for 22 h and the turbidity measured at 660 nm.
Results and Discussion The values for total folate, pantothenic acid and biotin as determined by microbiological assay in the
Folacin activity, ug/IOOg Range 5.2-10.2 10.1-13.3
Mean ± S.D.
± 5.2 ± 9.8
± ±
1.8 1.5
± 2.6
6.1-12.0
±
2.0
± 2.9 ± 8.6
7.6- 9.8 8.1-24.2
± 2.3
2.7- 8.5
± ± ± ±
1.0 0.7 2.8 6.7
2.4- 5.0 9.0-15.7 3.9-13.0 7.4-15.7
± ± ± ± ± ± ± ± ±
4.6 4.6 6.9
3.1-12.8 5.9-10.7 5.1-12.4 4.8-15.7 9.3-10.9 11.8-12.7 2.7- 6.6 2.4- 3.6 3.3- 5.3
I.I
0.2 0.6 2.2 2.2 3.7
8.2 11.1 2.9 8.9 8.0 8.6 16.1 9.0 6.3 6.6 3.4 13.6 7.7 10.7 17.8 8.0 8.5 8.3 10.2 10.1 12.2 4.6 3.0 4.3
± 1.1 ± 11.3 ±
3.2
± ± ± ±
1.4 2.8 3.8 4.4
± ± ± ± ± ± ± ± ±
4.2 2.3 3.7 7.8 1.1 0.6 1.7 0.9 1.4
different raw and processed fish products are shown on a fresh weight basis in Tables 1, 2 and 3, respectively. In general, where comparison with published levels of these vitamins in fish products was possible, the values compared favourably (Orr, 1969; Perloff and Butrum, 1977; Paul and Southgate, 1978). Although this study was not designed to measure vitamin losses due to preparation in the same sample of the different fish products, tentative estimates of lower cooking losses of folate, pantothenic acid and biotin have been reported for fish than for vegetables and meat (Paul and Southgate, 1978). In the case of canned samples
Table 2. Pantothenic acid in raw and processed fish products. Fish Product
Sample (n)
Cod
Haddock Halibut Lobster Pickerel Salmon Sardines Scallops Shrimp Trout Tuna Whitefish
-raw/frozen -baked -smoked -raw/frozen -baked -raw/frozen -raw/frozen -cooked -raw/frozen -raw/frozen -canned -canned -raw/frozen -cooked -raw/frozen -cooked -raw/frozen -smoked -baked -canned -raw/frozen -baked
Can. Insl. Food Sei. Teehnol. J. Vol. 22, No. 2, 1989
7 4 I 5 I 2 I I
4 2 4 4 I I
4 3 3 2 2 4 2 I
Moisture 010 Mean ± 5.0. 77.0 ± 5.5 68.2 ± 9.8 73.4 79.2 ± 2.6 75.2 76.0 ± 3.9 83.8 76.7 79.0 ± 2.3 65.7 ± I.I 67.8 ± 1.2 6.63 ± 3.1 82.9 72.5 85.5 ± 3.2 77.1 ± 2.1 69.8 ± 6.9 67.5 ± 0.2 67.9 ± 0.6 68.7 ± 2.4 75.2 ± 2.2 74.0
Pantothenic acid, ug/lOOg Range 189- 334 198- 276
Mean ± 5.0. ± ±
50 40
178- 251
±
30
±
10
297- 314 239- 628 977-1064 606- 828 432- 640 34- 106 82- 142 968-2326 1339-1346 1213-1642 167- 282 866-1209
256 223 227 221 247 305 2398 1041 356 1021 744 511 149 161 80 113 1724 1342 1427 218 1037 891
± 180 ± 60
± 100 ± 90 ± 30 ± 30 ± 690 ± 10 ± 300 ± 50 ± 240
Hoppner and Lampi / 171
Table 3. Biotin in raw and processed fish products. Fish Product
Cod Haddock Halibut Lobster Pickerel Salmon Sardines Scallops Shrimp Trout Tuna Whitefish
-raw/frozen -baked -smoked -raw/frozen -baked -raw/frozen -raw/frozen -cooked -raw/frozen -raw/frozen -canned -canned -raw/frozen -cooked -raw/frozen -cooked -raw/frozen -smoked -baked -canned -raw/frozen -baked
Sample (n) 7 4 I
5 1 2 1 1 4 2 4 4 1 1 4 4 3 2 2 4 2 1
the addition of vitamins through other ingredients could distort such estimates. The profile of folate derivatives and polyglutamate chain length has been reported for a few selected foods but not for fish (Rodriguez, 1978). It is presently not known whether some of the differences in vitamin content as indicated for pantothenic acid is species related, due to feeding habit or possibly commercial rearing. Data for the vitamin content of fish are scarce and not all foods have been analyzed for the above vitamins, particularly biotin. It is hoped that many of the gaps in data for these vitamins can be filled and thereby improve the data base of the Canadian Nutrient File.
Acknowledgements The authors thank the staff of the Total Diet Study for their technical assistance in the collection and preparation of the samples. References Baker, H. and Frank, O. 1967. A microbiological assay for folate activity. In: Vitamins, Chemistry, Physiology, Pathology, Methods. Vol. VII, 2nd ed. P. Gyorgy and W.N. Pearson (Eds.) p. 269. Academic Press, NY. Bonjour, J.P. 1985. Biotin in human nutrition. Ann. N. Y. Acad. Sci.447:97. Department of National Health and Welfare. 1983. Recommended Nutrient Intakes for Canadians. Bureau of Nutritional Sciences, Food Directorate, Health Protection Branch, Ottawa, Ont.
172 / Hoppner and Lampi
Moisture 070 Mean ± S.D. 77.0 ± 5.5 68.2 ± 9.8 73.4 79.2 ± 2.6 75.2 76.0 ± 3.9 83.8 76.7 79.0 ± 2.3 65.7 ± 1.0 67.8 ± 1.2 62.6 ± 3.1 82.9 72.5 86.7 ± 1.8 79.2 ± 4.6 69.8 ± 6.9 67.5 ± 0.2 67.9 ± 0.6 68.7 ± 2.4 75.2 ± 2.2 74.0
Biotin, ug/100g Range 1.1- 2.8 2.0- 3.2
2.0- 3.4 3.0-3.3 1.4- 2.5 6.7- 8.1 7.7-12.5 8.4-10.1 0.3- 0.7 0.5- 1.0 3.3- 6.5 4.0- 4.3 2.6- 3.6 1.1- 3.3 2.7-5.9
Mean ± S.D. 2.2 ± 0.6 2.6 ± 0.5 1.1 2.5 ± 0.5 2.5 3.1 ± 0.2 4.5 6.8 1.9 ± 0.6 7.4 ± 1.0 9.4 ± 2.1 9.1 ± 0.8 1.1 1.5 0.5 ± 0.2 0.8 ± 0.2 4.5 ± 1.7 4.1 ± 0.2 3.1 ± 0.7 2.1 ± 0.9 4.3 ± 2.3 4.2
Fox, H.M. 1984. Pantothenic acid. In: Food Science and Technol· ogy, Hand Book of Vitamins: Nutritional, Biochemical and Clinical Aspects. L.J. Machlin (Ed.). pA37 Marcel Dekker Inc., New York, NY. Herbert, V. and Bertino, J .R. 1967. In: Vitamins, Chemistry, Physiology, Pathology, Methods. Vol. VII, 2nd ed. P. Gyorgy and W.N. Pearson (Eds.). p. 243. Academic Press, New York, NY. Hoppner, K. and Lampi, B. 1982. Total folacin activity in breakfast cereals. Nutr. Rep. Int. 26:495. Orr, M.L. 1969. Pantothenic acid, vitamin B6 and vitamin B12 in foods. Home Econ. Res. Rep. 36, U.S.D.A., Washington, D.e. Paul, A.A. and Southgate, D.A.T. 1978. The Composition of Foods. R.A. McCance and E. Widdowson (Eds.). Her Majesty's Stationary Office, London, UK. Perloff, B.P. and Butrum, R.R. 1977. Folacin in selected foods. J. Am. Dietet. Assoc. 70: 161. Rodriguez, M.S. 1978. A conspectus of research of folacin requirements of man. J. Nutr. 108:1983. Skeggs, H.R. 1963. Biotin. In: Analytical Microbiology. F. Kavanagh (Ed.). pA21. Academic Press. New York, NY. Verdier, P. and Beare-Rogers, J.L. 1984. The Canadian Nutrient File. J. Can. Dietet. Assoc. 45:52. Wyse, B.W., Song, W.O., Walsh, J.H. and Hansen, R.G. 1985. Pantothenic acid. In: Methods of Vitamin Assay. J. Uugustin, B.P. Klein, D. Becker and B.P. Venugopal (Eds.). p. 399. John Wiley and Sons, New York, NY. Zook, E.G., MacArthur, J.M. and Toepfer, E.W. 1956. Pantothenic acid in foods. U.S.D.A. Agric. Handbook 97.
Submitted October 24, 1988 Revised December 20, 1988 Accepted January 2,1989
J. Inst. Can. Sci. Technol. Aliment. VD!. 22. No. 2. 1989