J. Sei. Fa Agie. 1978, 29, 1069-1075 With the Compliments of the Authise{ip Effect of Baking on the Nutritive Value of Pakistani Bread Mohammed Akmal Khan# and Bjorn O, Eggum Department of Animal Phsiolgy and Chemistry, Naional Intute of Animal Science Rolighedee} 28, DK-1988, Copenhagen, Denmark (Manuscript received 10 Api! 1978) ‘The nutritive value of Pakistani breads prepared from wheat, maize, rie, barley, millet, triticale and sorghum were measured chemically, by proximate, fatty acid and amino acid analyses and biologically in N-balance experiments with growing ruts ‘Yeast was not used in the preparation of doughs. The levels of lysine, threonine and tryptophan were in several cases negatively affected by the baking procedure. According to FAO 1973 scoring pattern, Iysine and threonine were found to be the first and second limiting amino acids in most of the breads. Methionine does not appear to be the thied limiting amino acid. Net protein utilisation was negatively affected by baking only in the ease of barley (6%) and maize (9%) whereas the bio logical value was improved in triticale (8°,) and sorghum (6%) compared to the corresponding unprocessed samples. The baking procedure did not affect the diges- tible energy in triticale. Net dietary protein calories percentage values indicate that the cereal breads, except sorghum, had values higher than 5%, 1. Introduction In Pakistan it has been reported that cereals provide over 72 and 76%, respectively, of the daily calories and protein intakes of an average person.! In certain areas, cereals constitute as much as '85% of the total caloric intake Wheat, in the form of bread, is by far the most common cereal product consumed and forms 88% of the total cereal intake.* Breads are also cooked from other ‘cereals and consumed in different seasons of the yea. Cereal protein is knowin to be of poor quality because ofits low content of certain essential amino acids, particularly lysine." Lysine and threonine were found to be the first and second limiting amino acids respectively for wheat,‘ barley. rce® and triticale.’ The sulphur containing amino acids have been shown to be the third limiting for wheat, corn’ and triticale.” Contrary to these reports it has been claimed? that the order of limiting amino acids in wheat protein is lysine, tryptophan, methionine, isoleucine and threonine. Although there is @ vast literature on the protein quality of flour and of yeast bread, there is still controversy regarding the influence of baking on protein quality. Eggum'® reported that baking caused partial reduction of lysine, tryptophan, methionine and cystine in European rye and wheat breads, whereas Clegg and Davis!! and Maleki and Djazayeri!® observed no change in the available lysine content of flour due to baking of European and Arabic wheat breads, respectively. Recently, Eggum and Duggal"? reported a $% reduction in the net protein utilisation of Indian wheat chapati as compared to wheat flour. On the other hand, Shyamala and Kennedy'* found the protein effi- ciency ratio of Indian chapati to be considerably higher than that of unprocessed wheat, Thus, there are conflicting reports with regard to second and third limiting amino acids in bres and the effect of baking on the protein quality of Indian, Arabic and European breads Since there is no published work on the protein quality of Pakistani bread made from cereals such as wheat, maize, rice, barley, millet, triticale and sorghum, the present work was initiated to Permanent address: Department of Nutrition, Unvenity of Agriculture, Faisal (0022-S142,78/1200-1069 $02.00 © 1978 Society of Chemical Industry1070 M.A. Khan and BO, Fu study the effect of baking without yeast on the nutritive value of Pakistani bread made from different cereals. Yeast is not commonly used in bread making in Pakistan 2. Experimental 2.1, Experimental animal and diets ‘The experimental procedure has been described by Eagum.!® Groups of five Wistar male rats, cach weighing approximately 75 g, were used. The preliminary period lasted for 4 days and the balance period for S days. The rats were weighed at the beginning of the experiments and divided into groups of five such that the average weights of the groups differed by no more than 0.5 g. Weighing was repeated at the end of the preliminary and balance periods: access to feed and water ‘was prevented 3 h before weighing. Each animal received 150 mg N and 10 g dry matter daily throughout the preliminary and balance periods. The N content of the diet was adjusted by using 1 basal diet consisting of a N-free mixture (Table 1). Me 1. Composition (pars by weigh) ofthe nitrogen-free mixture Potato sate (autocaved) 107 Svcrore % Callie powder 2 Soy bean ot 2 Mineral mixtures o YViumin misture” (mined with auoeaved potato starch) 20 “To provide per ke dit: CaCO, 2.74: callum cluae, CC HysO,c4H0, 12.53 8: CAHPOC2H.O, 431 g: KeHPOW 875 b KCI, 498 g: NaC, 08 g: MESO. 1-53 8: MECOs 1.41 g: ammo: nism frre cae (205-225, Fe), O41 g; MaSO- 1.0, 8.0 me CuSOUSH.O. St mg: KI, 1.6 mg: NaF, 20. mg: AINH ASO.) THs. 36. me "To prone pet ka dit etinol equivalent, 1.2 mg cholecakfra 2.5 na:thami, 8 mgs ibotacin, 2m colina, 8 mgs panto: ham ci, mg atocopherol, 4 mgs pyraloine, 0.2 ms ‘The experimental diets were weighed out into plastic boxes with tightly fitting lids for each of the preliminary and balance periods. The diet was weighed each day from these boxes in four daily allowances in the preliminary period and five daily allowances during the balance period. Any remaining diet was weighed and taken into consideration in the calculation of the experimental ‘Wheat, maize, rie, barley, millet (Baja), triticale and sorghum grains were ground to whole meal flours and breads were baked as follows: the doughs were first prepared by mixing the flour \with water (70% of wheat, barley and triticale flours; 75-80°% of rice, millet, maize and sorghum flours) and kneaded. Yeast was not added to any of the doughs. A portion of the dough was rolled fn a wooden board until it was Mat and circular. It was then placed on a heated flat iron plate at 220-230'C temperature. Wheat, barley and triticale breads were baked for 1.5-2 min while rice, millet, maize and sorghum breads were cooked for 7-10 min, All the bread samples were freeze dried, ground and incorporated into the test diets (Table 1), at the expense of autoclaved potato starch 2.2, Analytical methods ‘The chemical composition of the diets was determined according to standard methods.!® Acid hydrolysis followed by ether extraction was used for the estimation of fat according to the method (of Stoldt17 The caloric value ofthe diets and faeces was determined in an IK A-calorimeter (Adia- batic). Metabolisable energy (ME) of the diets was calculated according to Miller and Payne.'*[Nutr value of Paks tot ‘The fatty acids were methylated according to Metcalfe er al!* and analysed by gic. The liquid phase was 15% ethyleneglycol succinate supported on Chromosorb W (AW, DMCS) 80-100 mesh. Amino acid analyses were earried out according to Weidner and Eggum® and Eggum.*! Tannin ‘was estimated as described by Eggum and Christensen® and statistical analyses were based on _methods described by Snedecor.® Al the assays were performed in duplicate, 3. Results 3.1, Chemical composition ‘The values forthe chemical composition of different flours and the corresponding breads are shown in Tables 2 and 3. As recommended by Tkachuk, a factor of 5.7 was used for the conversion of nitrogen to protein, ‘Table 2 reveals thatthe protein content of various breads ranged from 6.9 to 14.5 ‘values were highest in maize and millet breads. Fibre contents were higher in barley and sorghum breads than in all other breads. Sorghum bread contained highest contents of ash (2.9%). Baking ‘of bread in general, increased the Values of calcium and sulphur whereas the phosphorus and “Table 2 Chemical composition (dry bas) of Nour and bread samples per 1008 Availabe (Calories per 100 ¢ re per 100.2 1008 Protein “earbo- Crude - (N37 Fat hysrate be Ash Toul ME* Co PS Fe Tannin Wheat four 2022 M1 12 16 4 4 a Hd RA 51 0.50 ‘Wheat bread Wa 2) 81 a Le a an 380 ss a03 Ss 0.3 lod S780 23 1.6 461 4 16K 0.7 2.6 42 O88 WS $2 WS 26 1S 489 42 IL USO Ma aS OS Rive Nour RI 14 901 0.7 06 42 40 125 1355 319 29 Om Rie bread 1 4 e407 0.6 40 a7 178 20s 3 25 OO” Brey Nour 439 649 43 22 4 4 @4 242 mS 70 Ot Barley bread SVR obs 44 22 451 Gn wd 393 ASR BI OT? Millet bread 122 54 711 18 17 488 482 30.3 335.6 43.3 6.2 0.62 Tritialefour =A 2.6 6742.7 20 AD AH. BL 2.9 0.70 Thiele bread «1.02.6 Oh LT 21 Me ADO 99.7 SSL O88 Sorghum flour «6.94.5 G74 4B 30 MT 2S 488 3.9 25.9 10.11.90 Sorghum brad «6.94.9 Gh SB O29 HO AM SNS BUh9 8.6 10.7 187 * ME, Metaboliable energy. Table 3, Fatty aid composition (s per 100g meures f) of Nour and bread Wheat Maze Rice Barley Milt —~—Triteale Sorghum Gc Flour Bread Flour Bread Flour Bread Flour Bread Flour Bread Flour Bread Flour Bread © 0 6 5 of oe 0 0 6 6 0 9 0 of 02 02 04 O41 19 18 08 07 OF 01 02 0.2 09 03 Ble Wh ASR 187 9 ae 27 264 mL MS RT dO 10 10 24 2 3411 42 29 29 09 10 14 20 99 1010 38.0 32 6 121 12.6 28.3 26.3 150 18 384 38.7 7 2 489 9 4 $43 S29 48 AL? 3 SHE A wD a7 4a 232 19 60 62 $0 49 60 $0 18 180a M.A, Khan ad B, 0. Eagum tannin contents were slightly reduced, Iron contents and fatty composition (Table 3) were not alfeted by baking of breads. 32, Protein quality Table 4 shows the total amino acid composition of flours and breads. The amino acid composition ‘of flour diflered from that of bread due to baking. Lysine, the most heat-sensitive amino acid was reduced slightly in the case of wheat and maize bread, 6% in barley bread and 7% in sorghum bread. “The content of threonine in various breads were also lowered slightly in mile, triticale and sorghum, 6-7% in rice and wheat, 18% in barley bread. The content of tryptophan seemed to be most affected and was lower by 8-10 in the case of millet, barley triticale and sorghum, 17% in maize and 27% in rice breads than those of respective flour values. Table 4, Amino acid companion (g per 16g N) of flour and bread Whee Maire Rice Barley Milet_—Tritiale Sorghum Amino acids Flour Bread Flour Bread Flour Bread Flour Bread Flour Bread Flour Bread Flour Bread Aspunivaid «4,6 «4S SS SB o8I AL SL 4d 67 66 59 59 6S 6S Threonine 2eo2@ $2 42 34 52 29 2S KI G4 29 Ze KS 32 Serine aa a3 a4 ak Ve 4a Xo Mr ak Ga Yo fo 4a 43 Glutamic acid 38.9 38.5 19.0 202 18.1 IKI 26.2 25.6 28.5 280 27.3 27.2 189 19.6 Proline for ws ‘83 89 43 42 107 le 70 69 8 Be 7S 79 Giyeine 37 36 34 34 82 41 36 RT NS RD SE aT BB 37 ‘amine aan 72 72 83 $2 99 37 62 62 X Ve ao 78 Isoleucine 3436 34 35 £0 39 35 34 3B 38 34 95 36 36 Leucine 69 68:27 28 77 76 67 67 #9 BR 64 64 2 IL Tyrowine 0 31 80 41 46 46 30 30 X00 30 28 29 46 Fs Phensialnine «4.7 4.7 43 46 48 47 49 49 43 G4 42 42 44 44 Uysine 24022 2s ae as Sa 4X2 M27 2's 2b alo 27 2's Wiaticine 2a 21 27 27 24 24 20 20 24 20 22 22 22 20 Arginine 22001 4 82 Te Ts 4 4G Us G3 SSE Te Lo Cystine 20020 20 19 20 20 20 18 19 17 20 20 22 23 Toppan «D0 08 OS TA OA TY NO ns 2 0 10 to Protein sore, based on the FAO scoring pattern (Table $) indicates that lysine isthe first limiting amino acid in all the lours and breads, threonine is the second limiting amino acid in wheat, rice, barley, millet, triticale and sorghum breads, whereas tryptophan was found to be the second limiting amino acid in maize bread. Isoleucine isthe third limiting amino acid in barley, millet and sorghum breads, valine in wheat and triticale breads, threonine and tryptophan in maize and rice breads respectively Results obtained on the effect of baking on true protein digestibility (TID), biological value (BV) and net protein utilisation (NPU) are summarised in Table 6. The TD of wheat, maize, rice and millet breads was not affected by baking, but was significantly lower in barley, triticale and sorghum breads compared with the corresponding flour values. The BY of maize and barley breads was significantly lover, while the BV of triticale and sorghum breads was significantly higher. The BV ff wheat, rise and millet breads was not differen from the unprocessed samples. NPU of triticale bread was significantly increased and was significantly lowered in ease of maize and barley breads from processing. The NPU values of wheat, rice, millet and sorghum breads were similar to theit unprocessed flours. The effect of baking on the digestible energy of cereals was tested by feeding ‘0 groups of ras on triticale lour and bread. No significant diflerence between the digestible energy of flour (87%) and bread (86%) was observedNitritve vale of Pakistan bread ‘nd tread samples Sample Wheat bread Rice our Rice bread arly flour Barley bread Mile four Mile eu Tale bread Sorghum four Sorghum bred be 6. Flt of baking onthe protein quality of Pakistan! breads 0 Wheat four 96.0 Wheat bread 95.0 Maize four v8.0 Rice bread tan.) Bary Nour 8.0 Barley bread $5.0" Mis our 93.0 Miles brea 92.0 Teitiale four 93.0 Trae bread 91.0" Sorghum bead $2.0" 0.08 sgnieauy dierent from coresponding value of four se s a Lysine Lysine Lysine Lysine Lysine Lysine Lysine Biologel —Necprowin Nev dieu ‘ale utbaion protein estore $5.0 0 37 seo See 59 61.0 $8.0 ss Soe Sor 32 10 10 3.6 m0. 22.0 83 0 56.0 63 a0 70 ea 210 sto ats imiting amino acids Second Third Threonine Valine Threonine Tryptophan Tryptophan Trronine Theonive Tryptophan Threonine Isoleucine Tiweonine Isoleucine Threonine Tnoeveine Threonine Valine "0.0 signtanily diferent from coresponding vale of flour, © P--0.001 since diferent From corresponding value of Hout [Net dietary protein calorie percentage (NDp eal were calculated according to Miller and Payne*® (Table 6). The NDp cal % ofthe breads lie between 3.3 and 86, The values of all the breads, except sorghum bread, were greater than $ NDp eal % 4. Discussion ‘Based on FAO/WHO 1973 scoring pattern, 073 of the different flour and bread samples “The most important factors determining the loss in nutritive value are the duration and temperature level of heat treatment and the levels of moisture and reducing substances.** “The processing conditions employed in the present work for preparing the different breads did rot decrease the nutritive value 10 a large extent, even in the samples (rice, millet, maize, sorghum)174 ML A. Khan and B. 0. Eggum exposed to 220-230'C for 7-10 min. However, making breads from barley and maize resulted in 6 and 9% decrease in NPU respectively, due to losses in the most limiting amino acids lysine, threonine and tryptophan. The results of the present studies, although the effect of processing was not so severe, agree with those found earlir.227 However, BV for rticale and sorghum breads were 8 and 6% higher respectively than those of Unprocessed samples. The moderate heat treaiment employed apparently had a positive influence fn the nutritive value due to destruction of antimetabolites. On the other hand the very high BV of sorghum is quite unrealistic, The sorghum used in this experiment was « local Pakistani variety having a dark brovin colour witha bitter taste, and one would have expected a high tannin content. Tannin estimated by the present method gave only 1.9% in the sorghum flour. This tannin value ‘cannot explain the very low digestibility values. However a specific analytical procedure has to be ‘undertaken to be able to analyse forall polyphenols in sorghum (Arora, S. K., personal communi tion), The present tannin values for sorghum are apparently far too low. Ifthe tannin content is higher than determined in the present work this might not only explain the low TD values but also the high BV values. As discussed previously. tannin has & highly negative efect on protein digestbi- lity, However, tannin reacts primarily with the non-essential amino acids proline, glutamic acid, slyeine and alanine. Consequently the BV of the absorbed protein must increase “The results also indicate that in the flours and breads of wheat and triticale, threonine is the second limiting amino acid and that valine is the third. Simvlar results have been reported for ‘whole wheat and wheat flour. Methionine does not appear to be the third limiting amino acid in the flour and bread samples. "According to FAO? the protein allowances for different age groups in terms of NDp cal % are 80, 7.8, 59, 8.4, 4.6 and 9.5 for infants, toddler, child, adolescent, adult and lactating mothers, respectively. All evaluated ceteal breads, except sorghum, give NDp cal values higher than 5% and should theoretically meet the protein requirement if consumed in adequate amounts. In general, processing of cereals into Pakistani bread (without yeast) affects the nutritive quality fonly to-4 minor extent. Acknowledgements ‘The authors are grateful to Miss I. Jacobsen, Mrs A. Tommerup and Miss M. Jensen for co: ‘operation and assistance during the course of research work References |. Minatey of Health, Government of Pakistan Nutrition Sariey of Pakiten Diretorate of Nutrition Survey Reearch, Islamabad, 1970 2. A'S. Mz Khan, M. A. Natron Sursey of Northern Arar of Pakistan University of Agriculture Pres Lyatpur, Pakistan, 1976 3. Khan M.A Alias, K': Abid, ARs Yagoob, M. Pakistan J. Agri. Sei 1976 13,167 4A. ander, ALE. Science 9.1. 1968, 127, 874 Home, EE Jamen, GR. Gillan, E,W. Am. J. Clin Nutr 1965, 16, 315. Rosenberg, H.R Cull, Rs Eckery RE, J. Nur. 1985, 6,217 3. Shimada, A. Cline, T. Re Anim Sei 74, 38, 94 5. Oewtormer, G. A Meade, R.J- Stockland, W.L.: Hanson, L. EJ. Anim, Se. 1970, 3161183, 9. King. K-W.: Sebel, W-H. 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