CHEMICAL COMPOSITION AND PROTEIN CHARACTERISTICS OF LENTILS

CHEMICAL COMPOSITION AND PROTEIN CHARACTERISTICS OF LENTILS R. S. BHATTY', A. E. SLINKARD', and F. W. SOSULSKI'z tcrp Develpment Centre and 2Crp Science Department, University f Saskatchewan, Saskatn, Sask. S7N 0W0. Received 27 Feb. 1976, accepted l7 May 1976. Can. J. Plant Sci. Dwnladed frm www.nrcresearchpress.cm by 37.44.193.220 n 02/09/18 Fr persnal use nly. Bunrrv, R. S., Sr-rNrARD, A. E. en Ssulsrr, F. W. 1976. Chemical cmpsitin and prtein characteristics f lentils. Can. J. Plant Sci. 56: '787-794. Chemical and amin acid cmpsitins, prtein slubility (Osbrn classificatin), and in vitr prtein quality f six gentypes f lentils (Lens culinaris Medik.) were determined. Lentils cntained an average f 28.6%, prtein, 3.l/a ash, 4.4Vc fiber, 0.77c ether extract, 63.lVc ttal carbhydrate (nitrgen-free extract), 44.39a starch and 4,186 kcal/kg fgrss energy. Lentil starch cntained 36.17c amylse. Abut ne-half (47.lqa) f the lentil prteins were sluble in l 0 M sdium chlride, 3.87 sluble in water, 3.lqr sluble in 7O% ethanl. and 14.9%,sluble in dilute hydrchlric acid. Only 24.UV f the lentil prteins were nt slubilized by these slvents. The lentils cntained 6.8Vc f the meal nitrgen as nnprtein nitrgen. The amin acid cmpsitins f the six lentil gentypes were similar. The majr amin acids were glutamic acid, aspartic acid, arginine, leucine and lysine; the minr amin acids were methinine, cystine and tryptphan. Because f the lwer cncentratins f the latter amin acids, lentil prteins gave a chemical scre f 35, a prtein scre f 46, and an essential arnin acid index f 63, relative t egg prtein. Les auteurs nt 6tabli la cmpsitin chimique 96n6rale, la cmpsitin aminacid6e, la slubilit6 des prt6ines (classificatin d'osbrn) et la qualit6 des prt6ines (in vitr) de six g6ntypes de lentille (Lens culinaris Medik.). Les lentilles cntenaient, en myenne, 28.6Va de prt6ine, 3.17c de cendre, 4.47c de cellulse, 0.7%, d'exrrait h l'6ther, 63.l%c d'hydrates de carbne ttaux, extrait nn azt6., 44.37c d'amidn et 4,186 kcal/kg d'6nergie brute. L'amidn se cmpse pur 36.lVc d'amylse. Envirn la miti6, 4'7.7%,, de la prt6ine est sluble dans une slutin 1.0 M de NaCl, 3.87a est sluble dans I'eau, 3.17c dans l'6thanl d,'7/r et 14.9%' dans HCI dilu6. Seulement 247c n'a pu 6tre mis en slutin par ces slvants. Envirn 6.87c de I'azte de la farine (chair) est sus frme nn prtdique. La cmpsitin aminacid6e a 6t6 la mome chez les six gentypes, les plus frtes cncentratins 6tant celles des acides glutamique et aspartique, de I'arginine, de la leucine et de la lysine, les plus faibles celles de la m6thinine, de la cystine et du tryptphane. La teneur plus faible de ces derniers dnne i la prt6ine de la lenrille un indice chimique de 35, un indice prt6ique de 46 et un indice d'acides amin6s essentiels de 63 par rapprt h la prt6ine de I'euf. Lentils (Lens culinarls Medik.) are under 1976). The 1975 lentil acreage in Saskinvestigatin as ne. f the alternate crps atchewan decreased t 150 ha. Lentil fr Western Canada. Mst f the Canadian prductin culd increase n the prairies if prductin f lentils has been cncentrated high-yielding and disease-resistant cultivars in Saskatchewan where field scale grwing became available. Lentils are largely grwn f lentils began in 1970 with 600 ha which n cntract fr exprt t Eurpe fr use in increased t 6,000 ha in 1972 (Slinkard human fd. Can. J. Plant Sci. 562 7t7-794 (Oct. 1976\ 787

788 CANADIAN JOURNAL OF PLANT SCIENCE Can. J. Plant Sci. Dwnladed frm www.nrcresearchpress.cm by 37.44.193.220 n 02/09/18 Fr persnal use nly. A number f studies have been reprted n the chemical cmpsitin f lentils. Saint-Clair (1972) reprted slubility f lentil prteins in different slvents (Osbrn classificatin). Khan and Baker (1957) and Bell (1975) reprted the chemical and amin acid cmpsitins f single gentypes f lentils. Anther publicatin (Harvey I97O) dcuments the amin acid cmpsitins f lentils grwn under widely different envirnmental cnditins. Hwever, there is paucity f infrmatin n cmpsitins and prtein quality f different gentypes f lentils grwn under cmparative cnditins. The bjectives f the present study were t determine the chemical cmpsitin, prtein slubility, amin acid cmpsitins and in vitr prtein quality f a number f lentil gentypes. MATERIALS AND METHODS Six gentypes f lentils which included bth the large-seeded Chilean types and the small-seeded Persian types were used in the study. The gentypes were riginally btained frm varius surces and grwn at Saskatn under cmparative cnditins in 1974.. Seeds were grund in a micrmill t pass a 1.0-mm screen. Misture and vlatile matter, prtein (Nx6.25), ash, fiber, and starch cntents f the meal were determined in duplicate by the fficial methds Assciatin f Official Analytical Chemists (AOAC) 1970. Ttal carbhydrate cntent (nitrgen-free extract) was calculated by difference. Amylse (idine affinity) was determined by the prtentimetric titratin methd f Schch (1964). Grss energy cntent f the meal was determined with a Gallenkamp adiabatic bmb calrimeter (Bhatty and Wu 1974). Nnprtein nitrgen (NPN) was determined by extracting I g meal with warm TOVI ethanl. The extract was centrifuged (5000 g; l0 min) t remve the insluble materials. Ethanl was remved frm the extract by rtary evapratin. The cndensed extract was made t vlume with distilled water and an aliqut taken fr the determinatin f nitrgen (AOAC 1970). Prtein slubility was determined by extracting at rm temperature (ca. 20 C) sequentially 5 s meal fr I h in a wrist-arm shaker with 75-80 ml each f 5.87 sdium chlride, '707a ehtanl, and 0.05 M hydrchlric acid. The extract was centrifuged (10,000 g; l0 min) in each case and aliquts f the supernatant fractins were taken fr the determinatin f nitrgen. The residual meal was dried vernight at 70 C and analyzed fr nitrgen. The sdium chlride extract was -dialyzed against several changes f distilled water until free f salt t separate the watersluble and -insluble (saltsluble) prtein fractins. The nn-dialyzable fractin was centrifuged (10,000S; l0 min) and an aliqut f the supernatant fractin analyzed fr nitrgen (water-sluble nitrgen) Fr amin acid analysis, except tryptphan, 30-35 mg f the meal was hydrlyzed, fr 24h at I l0 C in a sealed, evacuated glass tube with l0 ml f glass-distilled 5.7 M hydrchlric acid. The amin acids were determined with a Beckman 120 C amin acid analyzer (Spackman et al. 1958). Cystine was cnverted t cysteic acid and methinine t methinine sulfne after perfrmic acid xidatin f the meal prir t acid hydrlysis (Mre 1963), and eluted n a 52-cm clumn f the analyzer. Tryptphan was determined n a l0-cm clumn f the analyzer after barium hydrxide hydrlysis f the meal (Tkatchuk and Irvine 1969). The mean essential amin acid cmpsitin f the six gentypes was used t calculate chemical and prtein scres and essential amin acid index (EAAI) aicrding t Sheffner (1967). Chemical and prtein scres are percent ratis and EAAI is the gemetric mean f the ratis f the l0 essential amin acid cntents f an egg and f lentils. RESULTS AND DISCUSSION The seed yield f the six gentypes f lentils varied frm 1,516 t 1,956 kglha, prtein frm 27.7 t 3l.37, ash frm 2.9 t 3.37, fiber frm 3.5 t 4.97, ether extract frm 0.5 t O.97, starch frm 42.4 t 47.67, amylse frm 33.0 t 37.4c/ and grss energy frm 4,085 t 4.268 kcalikg (Table l). Bell (1975) reprted generally similar prtein, ash, fiber and nitrgen-free extract cntents f lentils. The studies by Bell (1975) and Khan and Baker (1957) did nt reprt data n starch, amylse and grss energy cntents f lentils. There were differences in all the chemical cmpnents

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790 CANADIAN JOURNAL OF PLANT SCIENCE Table2. Nnprtein nitrgen (NPN) and slubility f lentil prteins in different slvents 7 f ttal prtein Can. J. Plant Sci. Dwnladed frm www.nrcresearchpress.cm by 37.44.193.220 n 02/09/18 Fr persnal use nly. Gentype Teka Mrden Amasaya Eskiseher Pineer Red Slvenskamdra Mean NPNl (E) 6.6 5.5 '7.2 7.2 6.6 7.8 6.8 5.87c NaCl Water- insluble Watersluble 4.9 4.4 3.7 3.9 3.0 3.8 55.0 48.2 4)) 50.7 48-3 42.1 fextracted separately and nt included in recvery. 47.7 between sme f the gentypes, thugh larger differences were btained in yield, prtein and fiber cntents than in ther cmpnents. The large-seeded gentype Teka (1,OO0-kernel weight, 76 g), the nly cultivar develped in Nrth America, and the small-seeded gentype Slvenskamdra (1,OO0-kernel weight, 30 g) had different seed yields but the same chemical cmpsitins. Gentypes Amasaya, Mrden and Eskiseher ( 1,00O-kernel weight, 52-54 g) had the highest yields and prtein cntents (except Mrden), and therefre wuld be better suited fr grwing under Saskatchewan cnditins. As in faba beans (Vicia faba L.) (Bhatty 1974), the prteins and ttal carbhydrates cmprised n a dry basis abut 927 f the ttal seed weight in lentils. Lentils cntained 42.4-45.OV starch cmpared t 46.3-55.97 in peas (Pisum sativum L.) (Kistra 1974), and 28.1-42.6q in faba beans (Bhatty 1974). The amylse cntent (36.V) f lentil starch was slightly lwer than the amylse cntents (38-40q) f peas and faba bean starches. The grss energy cntent f lentils (4,186 kcallkg) was similar t that f peas and faba beans reprted by Bhatty and Wu (r97 4). Abut 697 f the ttal lentil prteins were sluble in the three slvents listed in Table 2 and, with the residual meal prteins, ttal prtein recvery was93.57. TOVa ethanl 2.) 2.5 3.1 3.2 J.J 3.5 3.1 0.05 M HCI t4.7 15.4 16.0 t3.4 13.2 16.7 t4.9 Residual Ttal prtelns recvery (E) (7) 18.9 21.5 26.2 t9.7 25.5 26.5 24.0 96.0 98.0 9t.2 90.9 93.9 91.7 93.5 The remaining 6-7V prtein was prbably lst as a dialyzable fractin during dialysis f the sdium chlride extract. The majr prtein fractin f lentils, like that f ther legumes, was the salt-sluble r waterinsluble fractin (glbulins) which frmed an average f 477 f the ttal seed prteins. The quantity f this fractin was smewhat lwer in lentils than the range f 50-757 reprted in ther legumes by Sun and Hall (1975). Mlina et al. (1914) reprted maximum prtein slubilities f apprximately 55V with 0.5 M sdium chlride,ph 7-12 in Jack beans (Canavalia ensifrmis); ur studies indicated that the slubility was highest at 1.0 M, ph 7.0 (Table 3). In additin t the glbulins, lentils cntained 3.8% water-sluble prteins (albumins), 3.lV ethanl.sluble prteins (prlamines, mst prbably lw mlecular Table 3. Slubility f lentil prteins in different cncentratins f sdium chlride (Slvenskamdra lentil) Salt cncenrailn (sc) t.2 2.3 3.5 4.6 ).6 Mlar.2.4 0.6 0.8 1.0 % fttal prtein J).J 41.4 40.9 4t.9 45.3

BHATTY ET AL.-COMPOSITION OF LENTILS '791 Can. J. Plant Sci. Dwnladed frm www.nrcresearchpress.cm by 37.44.193.220 n 02/09/18 Fr persnal use nly. weight nitrgenus cmpunds), and 14.9V hydrchlric acid-sluble prteins (glutelins). Abut a quarter (24.OV) f the meal prteins were nt slubilized by the slvents. Saint-Clair (1972) reprted that lentils cntained abut 26V albumins,44v glbulins, 2.}Va prlamines and 207 glutelins n an 8.07 misture basis. The mair difference between the prtein stubility data reprted by Saint-Clair (1972) and that in the present study was in the water-sluble prtein (albumins) fractin. Faba beans cntain 6-77 f the meal prteins as albumin prteins (Bhatty, R. S. unpublished data), and Grant et al. (1974) reprt that peas cntain abut l57 albumin prteins. Hwever, the quantity f albumin prtein in a legume seed may depend, in part, n the methd f extractin. Direct extractin f the meal with water slubilizes, in additin t the albumin prteins, sme nnprtein nitrgen and may slubilize sme salt-sluble prteins due t the frmatin f weak salt slutin. Exhaustive extractin f the meal with a salt slutin fllwed by precipitatin f the salt-sluble prteins by dialysis gives a better estimate f this prtein fractin (Bhatty 1976). Lentils cntained 6.87a f the meal prteins as NPN. The NPN is a variable fractin and depends n the methd and slvent used in extractin (Bhattv and Finlaysn 19731. The quantiry f NpN extracted frm lentils was similar t that f faba beans (Bhatty, R. S. unpublished data), but higher than that f peas (Bhatty et al. 1973) extracted by a similar prcedure. The six gentypes f lentils had almst identical amin acid cmpsitins (Table 4). A lw cncentratin f ammnia in the hydrlysates indicated a lw level f amide nitrgen in the meal. The majr amin acids f lentils were glutamic acid, aspartic acid, arginine, leucine and lysine. These amin acids frmed mre than half f the ttal amin acids. Methinine, cystine and tryptphan were present in relatively lwer cncentratins. All the gentypes cntained an unknwn amin acid which was cnsistently eluted between ammnia and arginine. N attempt was made t identify this amin acid. The amin acid cmpsitin f lentils was similar t that f faba beans reprted by Kaldy and Kasting (19'74) and t the essential amin acid cmpsitins f lentils. sybean meal (Glycine max Men.) field peas, faba beans and field bean (Phaselus vulgaris) reprted by Bell (1975), except that beans cntained less arginine than lentils. Khan and Baker (1957) reprted higher levels f lysine, aspartic acid, glutamic acid, leucine and isleucine in lentils than thse fund in the present study. Hwever, these differences may, in part, be due t different methds used fr amin acid analysis. The gentype means f the l0 essential amin acids were taken fr the evaluatin f quality f lentil prteins relative t the quality f egg prtein. This evaluatin f lentil prtein gave a chemical scre f 35, a prtein scre f 52, and an EAAI f 63 (Table 5). The chemical and prtein scres suggested that the first limiting amin acids f lentils, like ther l6gumes (Bandemer and Evans 1967). were methinine and cystine, and the next limiting amin acids were tryptphan and isleucine. Based n these scres, the quality f lentil prteins was nly half t ne-third that f egg prtein. The EAAI f 63 als suggests that the quality f lentil prtein is lw cmpared t an EAAI f 100 fr egg prtein, 91 fr casein and 67 fr whle wheat (Osler 1970). Sarwar et al. (1975) reprted a chemical scre f 30 fr faba bean and 36 fr peas, and Kaldy and Kasting (1974) reprted an average prtein scre f 40 fr faba beans. Hwever, these scres were calculated relative t the essential amin acid requirements f the rat rather than t the essential anin acid cmpsitin f'an egg as has been dne in this study. In ur view, selectin f lentil fr prtein quality is nt warranted, as chemical and prtein scres f lentils may be greatly imprved, like thse f peas and faba bean (Sarwar et al. 197 5), by supplementing with methinine.

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BHATTY ET AL.-COMPOSITION OF LENTILS 793 Table 5. Chemical and prtein scres, and essential amin acid index (EAAI) f lentils Can. J. Plant Sci. Dwnladed frm www.nrcresearchpress.cm by 37.44.193.220 n 02/09/18 Fr persnal use nly. Essential amin acid Lysine Tryptphan Isleucine Valine Arginine Methinine & cystine Threnine Leucine Phenylalanine & tyrsine Histidine Ttal Egg'l' Lentilsd (g/16 g nitrgen) 6.6 l-7 6.6 7.4 6.6 5.4 5.0 8.8 10.8 2.4 61.3 ttaken frm Sheffner (1967) fmean f six determinatins (see Table 4)..J 0.9 3.5 4.0 6.9 1.9 J. I 6.4 6.4 )1 41.5 ACKNOWLEDGMENTS We are indebted t Dr. J. Vse, Prairie Resinal Labratry, Saskatn fr the determinati-n f amylse, and t K. Wu and H. Braitenbach fr technical assistance. ASSOCIATION OF OFFICIAL ANALYTICAL CHEMISTS. 1970. Official methds f analysis. I lth ed. AOAC, Washingrn, D.C. BANDEMER, S. L. and EVANS, R. J. 1967. Nutritive value f legume seed prteins. J. Agric. Fd Chem. 15: 439-443. BELL, J. M. 19'75. IJtllizarin f prtein supplements in animal feeds. Pages 633-656 in J. T. Harapiak, ed. Oilseed and pulse crps in western Canada. Mdern Press, Saskatn, Sask. BHATTY, R. S. 1974. Chemical cmpsitin f sme faba bean cultivars. Can. J. Plant Sci. 54: 4t3-421. BHATTY, R. S. and WU. K. K. 1974. Determinatin f grss energy f cereals and legumes with a ballistic bmb calrimeter. Can. J. Plant Sci. 54: 439-441. BHATTY; R. S., SOSULSKI. F. W. and WU. K. K. 1973. Prtein and nn-prtein nitrgen cntents fsme ilseeds and peas. Can. J. plant Sci.53:651-657. BHATTY, R. S. and FINLAYSON. A. J. t9i3. Extractin f nn-prtein nitrgen frm ilseed meals with diff'erent slvents. Cereal Chem. 50: 329-336. GRANT, D. R., SUMNER. A. K. and JOHNSON, J. M. 1974. Investiearins f the Chemical scre 9J 53 )-J l4 35 62 73 59 8'l Prtein scre t4l 79 78 "19 t54 52 93 108 8'7 130 EAAI (Lg fc ratis) 1.98 l.'72 1.72 t. tj 2.OO 1.54 t? l.86 1.7'7 t.94 Average lg 1.81 Antilg (EAAI) 63 albumin fractins f pea seed prteins. Abstr. 126. Amer. Assc. Cereal Chem. Cnf., Mntreal, Que. HARVEY, D. 1970. Tables f the amin acids in fds and feeding stuff. Tech. Cmm. N. 19. 105 pp. Cmmnwealth Bureau f Animal Nutritin. Bucksburn, U.K. KALDY, M. S. and KASTING, M. t974. Amin acid cmpsitin and prtein quality f eight faba bean cultivars. Can. J. Plant Sci. 54: 869-87 l. KHAN, N. A. and BAKER, B. E. 1957. The amin acid cmpsitin f sme Pakistani pulses. J. Sci. Fd. Agric. 8:301-304. KOOISTRA, E. 1974. On the differences between smth and three types f wrinkled peas. Euphytica ll: 357-3'l 3. MOLINA, M. R., ARGUETA, C. E. ANd BRESSANI, R. 1974. Extractin f nirrgenus cnstituents frm the Jack bean (Canavalia ensifrmis). J. Agric. Fd Chem. 22:309-312. MOORE, S. 1963. On the determinatin f cystine as cysteic acid. J. Bil. Chem. 238: 235-237. OSLER, B. L. l9'7o. Assessment f the nutritinal value f nvel prteins. J. Amer. Oil Chem. Sc.47:453-457. SAINT-CLAIR, P. M. 1972. Respnses f Lens esculenta Mench t cntrlled envirnmental factrs. Cmm. Agric. Univ. Wageningen. The Netherlands. pp. 72-77. SARWAR, c., SOSULSKI, F. W. and BELL, J. M. 1975. Nutritive value f field pea and fababean prteins in rat diets. Can. Inst. Fd Sci. Tech. J. 8: 109-l 12.

794 CANADIAN JOURNAL OF PLANT SCIENCE Can. J. Plant Sci. Dwnladed frm www.nrcresearchpress.cm by 37.44.193.220 n 02/09/18 Fr persnal use nly. SCHOCH, T. 1964.ldimetric determinatin f amylse. Pages 157-160 in R. L. Whistler, ed. Methds in carbhydrate chemistry. Academic Press, New Yrk, N.Y. SHEFFNER, A. L. 1967. In vitr prtein evaluatin. Pages 125-195 in A. A. Albanese, ed. Newer methds f nutritinal bichemistry. Academic Press, New Yrk, N.Y. SLINKARD, A.E. 19'16. Field peas, field beans, lentils. Agrlgist 5(2): 15, 17. SPACKMAN, D. H., STEIN, W. H. and MOORE, S. 1958. Autmatic recrding apparatus fr use in the chrmatgraphy f amin acids. Anal. Chem.30: ll90-1206. SUN, S. M. and HALL, T. C. 1975. Slubility characteristics f glbulins frm Phaselus seeds in regard t their islatin and characterizatin. I. Agric. Fd Chem. 23: I 84- I 89. TKATCHUK, R. and IRVINE, G. N. 1969. Amin acid cmpsitin f cereals and ilseeds. Cereal Chem. 46: 206-218.

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