J Sci Food Agric 1998, 76, 404È410 Study of the Nutritional Value of Black Cumin Seeds (Nigella sativa L) Hamed R H Takruri* and Majdoleen A F Dameh Department of Nutrition and Food Technology, Faculty of Agriculture, University of Jordan, Amman, Jordan (Received 6 November 1995 ; revised version received 20 May 1997 ; accepted 9 July 1997) Abstract : The nutritional value of Ðve samples of black cumin seeds from Ðve di†erent sources available in the local market were studied. The average values of the proximate analysis on dry matter basis were 216 g kg~1 for crude protein, 406 g fat, 45 g ash, 84 g crude Ðbre and 249 g of nitrogen-free extract, whereas moisture content was 38 g kg~1. The mineral and vitamin analyses showed that black cumin seeds contained iron, (105 mg kg~1) copper (18 mg), zinc (60 mg) phosphorus (527 mg), calcium (1860 mg), thiamin (15É4 mg), niacin (57 mg), pyridoxine (5É0 mg) and folic acid (160 kg). The protein quality of black cumin seeds was evaluated using net protein utilisation (NPU), protein efficiency ratio (PER) and net dietary protein energy percent (NDPE %) for two samples imported from Syria and Turkey, while PER was determined for the Syrian sample only. The results indicated that the NPU standardised of Turkish black cumin seeds was signiÐcantly higher than that of Syrian type (P \ 0É05). The mean results (^SD) were 54É6 ^ 2É72 for the Syrian type and 63É1 ^ 3É74 for the Turkish type. The NDPE% mean results (^SD) were 5É3 ^ 0É79 and 5É6 ^ 0É26 for the Syrian and the Turkish samples, respectively. The PER adjusted value for the Syrian samples was 1É9. The results of protein quality evaluation and those of the nutrient composition suggest that black cumin is of relatively good nutritional value. ( 1998 SCI. J Sci Food Agric 76, 404È410 (1998) Key words : nutritional value ; black cumin ; Nigella sativa ; protein evaluation ; net protein utilisation ; protein efficiency ratio and diuretic agent (Babayan et al 1978 ; Ustum et al 1990). Recently, many medical properties have been attributed to the black cumin seeds and/or its oil, including antineoplastic (antitumour) (Salomi et al 1992), antibacterial (Hasan et al 1989 ; Hussein, 1990 ; Hanafy and Hatem, 1991), antifungal (Hussein, 1990) and antihelmenthic (Salomi et al 1992) e†ects. Further studies indicated the presence of bronchodialator, immunopotentiating and hypotensive activities (Aqel 1992a,b). Also, antispasmodic, hypotensive glycosides and hypertensive alkaloids were isolated from the seeds of this plant (Hasan et al 1989). The expressed oil has been used for treatment of asthma (Babayan et al 1978). More recently, a great deal of attention has been given INTRODUCTION Black cumin (Nigella sativa) is an annual spicy, dicotyledon of the Ranunculaceae herbaceous plants growing in countries bordering the Mediterranean Sea (Gad et al 1963). The seeds are used for edible and medicinal purposes in many countries including India, Egypt and Syria. They are used in the preparation of a traditional sweet dish, composed of black cumin paste which is sweetened with honey or syrup, and in the preparation of black cumin pastry. They are also used for sprinkling on bread, Ñavouring of foods, especially bakery products and cheese, and as a carmanitive, stomachic * To whom correspondence should be addressed. 404 ( 1998 SCI. J Sci Food Agric 0022È5142/98/$17.50. Printed in Great Britain Nutritional value of black cumin seeds 405 to the black cumin seeds and their oil, and thus their consumption has increased especially in Middle East countries. Work on the nutritional value is scarce. There are few studies on the oil characteristics and fatty acid composition of black cumin (Gad et al 1963 ; Babayan et al 1978 ; Ustum et al 1990). Babayan et al (1978) studied the proximate analysis, fatty acid and amino acid composition of the black cumin seeds and found that the seeds are rich in crude fat, polyunsaturated fatty acids (particularly linoleic and oleic) and protein. The amino acid composition showed that the two amino acids usually limiting in plant foods, lysine and methionine, are high. Thus, it was thought important to conduct the present study, which aimed at determination of the proximate analysis, study of the protein quality and measuring of some vitamins and mineral contents in black cumin seeds. MATERIALS AND METHODS Nutrient analysis Five samples of black cumin seeds were purchased from di†erent local groceries. These samples were reported to be imported from India (one sample), from Syria (two samples), from Turkey (one sample) or grown in Jordan (one sample). Total nitrogen content was determined by the standard Kjeldahl method and total fat by Soxhlet extraction of the ground seeds for 24 h using petroleum ether (40È60¡C). Moisture, ash and crude Ðbre were determined by standard AOAC (1990) procedures. Mineral analysis was done on the hydrochloric acid solution of the ash. Calcium, sodium, copper, iron, zinc and potassium were determined by atomic absorption spectrophotometry (Pye Unicam) and phosphorus by (Uvikon 810 spectrophotometer) according to the AOAC. Vitamin analyses were done using high performance liquid chromatography (HPLC) according to the method described by Lam et al (1984). The proximate analysis of the diets used in animal experimentation (NPU and PER), presented in Table 1, was done according to methods described in AOAC (1990). Net protein utilisation determination Protein quality was assessed using net protein utilisation (NPU) and protein efficiency ratio (PER). The NPU procedure used was that of Miller (1963) with some modiÐcation. The breeding colony of rats suggested by Miller was not available, and weanling rats of the Sprague Dawley strain, 23 days old, were used. They were fed a stock diet for 1 week and then divided into four groups of Ðve rats each. The di†erences in mean weight between any two groups did not exceed 2 g. Each rat was individually caged and maintained at a temperature of 26 ^ 2¡C. One group was given a casein diet, the second was fed a protein-free diet (PFD), and the other two groups were fed the experimental diets containing black cumin (Syrian and Turkish types) ground to pass through a 1 mm sieve. The composition of diets is shown in Table 2. The fat of black cumin seeds used in the NPU experiments was partially extracted with petroleum spirit (40È60¡C) ; the mixture of ground seeds and solvent was stirred several times, allowed to settle for 6 h, decanted Ðltered and the residue oven-dried overnight at 75¡C. Two similar experimental runs were conducted under the same conditions 6 months apart. The diets were fed for 10 days in each of the two runs. The animals were weighed and killed by chloroform anesthesia. The abdomen, thorax and skull of the animals were opened before they were dried in a hot-air oven set at 105¡C for 48 h. Body water was calculated and carcass nitrogen was determined by the Kjeldahl method. The NPU operative, TABLE 1 Proximate analysis of the diets used in the NPU and PER experiments (g kg~1 wet matter basis)a Diets Moisture Ash Crude protein Ether extract Crude Ðbre Nitrogen-free extractb NPU experiment Syrian black cumin Turkish black cumin Casein Non-protein 88 82 94 102 55 55 40 31 111 91 96 04 124 131 92 80 69 41 NDd NDd 553 600 678 783 3773 3935 3920 3862 88 93 34 47 118 118 197 211 37 NDd 526 531 4345 (18É19) 4491 (18É80) PER experiment Syrian black cumin Casein a b c d Calories (kcal (MJ))c (15É79) (16É47) (16É41) (16É17) Mean of triplicate with CV \ 5%. The values of nitrogen-free extract are calculated by di†erence. Calculated by multiplying grams of crude protein and nitrogen-free extract by 4 kcal and ether extract by 9 kcal. Not detected. H R H T akruri, M A F Dameh 406 TABLE 2 Composition of the diets used to feed the rats in NPU and PER experiments (g kg~1) Ingredient NPU experiment Casein Corn starch Corn oil Vitaminised carbohydratea Salt mixtureb Fat ] fat soluble vitaminsc Test food PER experiment Casein diet Non-protein diet Black cumin (Syrian type) diet Black cumin (T urkish type) diet Casein diet Black cumin (Syrian) diet 113É8 737É9 88É3 10 40 10 È È 850 90 10 40 10 È È 511 44 10 40 10 385d È 526 28É4 10 40 10 385d 113É8 651É7 174É5 10 40 10 È È 447É4 È 10 40 10 492É6 a The vitaminised carbohydrate contained 45 g ascorbic acid, 0É06 g thiamine hydrochloride, 1É2 g calcium pantothenate, 4É0 g nicotinic acid, 4É0 g inositol, 12É0 g p-aminobenzoic acid, 0É04 g biotin, 0É04 g folic acid, 0É001 g cyanocoblamin and 12É0 g choline chloride, made up to 1 kg with corn starch. b The salt mixture is made by mixing 0É21 g Al (SO ) 7K SO 24H O, 309É85 g CaCO , 300 g KH PO . 3H O, 0É26 g 2 4 2 4 2 3 2 4 2 CoCl . 6H O, 0É5 g CuSO . 5H O, 23É56 g Fe (SO ) . 7H O, 51É13 g MgSO , 4É13 g MnSO , 0É83 gKI, 135É48 g, 2 2 4 2 2 43 2 4 4 K HPO . 3H O, 173 g NaCl, 0É26 g NaF, 0É26 g Na B O . 10H O and 1É32 g ZnSO . 7H O. 2 4 2 2 4 7 2 4 2 c Fat-soluble vitamins mixture is made by mixing 18 000 IU vitamin A, 2000 IU vitamin D and 500 mg a-tocopherol 2 per gram of fat. d Fat-extracted black cumin containing \10% fat in NPU experiment. NPU(op), was calculated using the Miller equation (Miller 1963) : NPU(op) \ (Carcass nitrogen of experimental group [ Carcass nitrogen of group on PFD) ] 100 Nitrogen intake The NPU standardised NPU(st), was also calculated using the equation of Miller and Payne (1961) : NPU(st) \ 54 ] NPU(op) [8 54 [ PE% where PE% is the energy value of protein in the diet as a proportion of total metabolic energy (Miller and Payne 1959). The caloric content was calculated using proximate analysis value of the diets and using the Ðgures 4, 9 and 4 kcal g~1 (16É7, 37É7 and 16É7 kJ) for carbohydrates, fats and protein, respectively. Net dietary protein as a percentage of total energy (NDPE %), which is an indicator of both quality and quantity of protein, was calculated by substituting the N%, NPU(op) and metabolisable energy (ME) values in the following formula (Miller, 1963) : NPU(op) 4 ] 6É25% NDPE% \ ] 100 kcal g~1 Protein efficiency ratio determination The PER method was that of Campbell (1963). The animals used were 23-day-old male weanling rats of Sprague-Dawley strain. They were divided into two groups. Each rat was housed in a single cage, and the temperature of the room was set at 26 ^ 2¡C. One group was given a casein diet, whereas the other group was fed an experimental diet (Syrian black cumin). The composition of the casein and black cumin diets are shown in Table 2. The diets were fed for 28 days, and the weight gain and food consumption were recorded every 3È4 days until the end of the study. The calculation of the PER, according to Campbell (1963), was as follows : PER \ Gain of test animal (g) Protein consumed (g) Statistical analysis Result of food proximate analysis, mineral contents, vitamin contents and animal experimentation (NPU and PER) were statistically analysed using DuncanÏs multiple-range test. RESULTS Nutrient composition of black cumin seeds Proximate analysis of the Ðve di†erent black cumin samples is presented in Table 3. The average results are compared with the reported values from Babayan et al (1978). Nutritional value of black cumin seeds 407 TABLE 3 Proximate analysis of Ðve black cumin sources (g kg~1 dry matter basis)a Seed source Moisture Ash Protein Ether extract Crude Ðbre Nitrogenfree extractb Energy (kcal (MJ)) PE%c Indian Jordanian produced Syrian 1 Syrian 2 Turkish Mean ^ SD 36 48 241 397 73 241 5501 (23É03) 175 40 36 37 42 38É3 ^ 2É7 42 47 45 44 45É2 ^ 2É4 199 209 225 207 216É2 ^ 16É8 401 401 425 406 406É0 ^ 11É1 75 129 85 58 84É0 ^ 26É9 283 214 220 285 248É6 ^ 33É8 144 158 161 147 157 ^ 12É4 55 38 213 353 55 340 5537 (23É18) 5301 (22É19) 5605 (23É46) 5622 (23É52) 5514É2 ^ 128É5 (23É08 ^ 0É50) 5407 (22É63) Babayand a b c d 158 Mean of triplicate with CV \5%. The values for nitrogen free extract are calculated by di†erence. PE% is the energy value of protein in the diet as a proportion of total metaboic energy (Miller and Payne 1959). Babayan et al (1978). Table 4 presents a comparison between black cumin and selected nuts and oil seeds regarding crude protein, fat, carbohydrates, crude Ðbre, ash and energy. Black cumin seeds seem to have similar protein and ash contents, lower fat but much higher crude Ðbre contents. Table 5 shows the contents of seven mineral elements (macro and micro) and four vitamins in black cumin seeds. The index of nutritional quality (INQ) values of black cumin seeds, which is the mathematical expression of TABLE 4 Proximate analysisa of black cumin seeds in comparison with selected nuts and oil seeds, calculated on dry matter basis (kg~1) T ypes of seeds Crude protein (g) Fat (g) Carbohydrate (g) Crude Ðbre (g) Ash (g) Almonds Pistachio nuts Sesame seeds SunÑower seeds Black cumin seeds 195 213 212 266 216 567 572 545 479 406 177 165 147 154 249 28 20 53 59 84 32 29 43 43 45 Energy (kcal (MJ)) 6750 6770 6590 6260 5510 (28É26) (28É34) (27É59) (26É20) (23É06) a Pellett and Shadarevian (1970). TABLE 5 Mineral and vitamin contents of black cumin seeds from Ðve di†erent sources (mg kg~1)a Seed source Indian Jordanian produced Syrian 1 Syrian 2 Turkish Mean ^ SD Minerals V itamins Fe Cu Na K Ca Zn P B1 B6 Niacin Folic acid 102 24 550 5517 1932 62 5043 13 04 48 700 107 93 91 130 105É0 ^15É6 18 17 15 18 18É4 ^3É4 419 535 535 440 496É0 ^61É3 4423 5606 5359 5380 5257É0 ^477 1867 2005 1946 1544 1859É0 ^182É7 59 59 66 56 60É4 ^3É8 5023 5769 5221 5267 5265É0 ^301É6 14 13 18 15 14É6 ^2É1 15 04 06 04 06É6 ^4É8 33 97 NDb 48 56É5 ^27É9 400 870 630 470 614 ^186É9 a Mean of duplicate samples (minerals) and triplicate samples (vitamins) with CV \5% ; values are given on dry matter basis. b Not determined. H R H T akruri, M A F Dameh 408 the nutritive value of food in relation to caloric and nutrient needs, was calculated. The INQ values presented in Table 6 show that black cumin seeds provide an abundance of protein, iron, copper, phosphorus, zinc, thiamin, pyridoxine and niacin. loss among rats of the group fed the non-protein diet. The results of NPU, both operative and standardised, and NDPE% are shown in Table 8. The NPU and NDPE% values of casein were higher than those of the other diets (P \ 0É05) ; the values for Turkish black cumin diet were higher than that for Syrian black cumin diet (P \ 0É05). ANIMAL EXPERIMENTATION Net protein utilisation experiment Protein efficiency ratio experiments The food consumption and weight gain data combined from the two runs of the NPU experiment are presented in Table 7. It is clear that the group of rats fed the casein diet had higher weight gain than any other group as related to their food consumption. The groups fed the test diet had lower weight gain, although their food consumption was higher. There was an expected weight The food consumption of the group of rats fed the casein diet was found to be higher than that of the group fed the Syrian black cumin diet. This was reÑected on the weight gain (see Table 9). PER results are also shown in Table 9. The PER value for black cumin diet was 1É9 when adjusted for a PER value of 2É5 for casein. TABLE 6 Index of nutritional quality (INQ)a for black cumin seeds Nutrient Average contents of black cumin per 100 g on wet basis US RDAb % of US RDAc INQ%d Energy (kcal (MJ)) Protein (g) Thiamin (mg) RiboÑavin (mg) Pyridoxine (mg) Niacin (mg) Calcium (mg) Iron (mg) Copper (mg) Zinc (mg) Phosphorus (mg) Folacin (mg) 531 (2É22) 20É8 1É5 0É1 0É5 5É7 185É9 10É5 1É8 6 526É5 0É061 2300 (9É63) 65 1É5 1É7 2 20 1000 18 2 15 1000 0É4 23É1 32 100 5É9 25 28É5 18É6 58É3 90 40 52É7 15É3 1 1É4 4É3 0É3 1É1 1É2 0É8 2É5 3É9 1É7 2É3 0É7 a Index of nutritional quality (INQ) \ Percentage of nutrient allowance (Guthrie 1989). Percentage of energy requirement b US RDA : United States recommended dietary allowances per day (Williams 1993). c The % of US RDA for 100 g of black cumin. d The rating of the nutritive quality is as follows : \0É5 for poor, 0É5È0É89 for fair, 0É9È1É5 for adequate, 1É51È4É9 for good and [5 for excellent. TABLE 7 Average feed intake and weight gain of rat groups for the Ðrst and second NPU runs (X1 ^ SD)a Rats group fed on Casein Turkish black cumin Syrian black cumin Non-protein diet Feed intake (g per rat per day) W eight gain (g per rat per day) 1st run 2nd run 1st run 2nd run 7É8 ^ 1É16 8É1 ^ 0É93 7É8 ^ 0É95 4É4 ^ 0É93 6É2 ^ 2É35 8É6 ^ 6É8 8É8 ^ 0É89 6É5 ^ 1É09 1É1 ^ 7É8 1É0 ^ 6É4 0É7 ^ 6É1 [2É2 ^ 5É2 1É1 ^ 7É5 0É8 ^ 6É0 1É0 ^ 7É0 [1É8 ^ 1É8 a Five weanling Sprague-Dawley rats per group were individually fed for 10 days. Nutritional value of black cumin seeds 409 TABLE 8 Net protein utilisation operative, (NPU(op)), net protein utilisation standardised (NPU(st)) and net dietary protein energy % (NDPE %) (X1 ^ SD)a T ypes of diet Syrian black cumin diet Turkish black cumin diet Casein diet NPU(op) NPU(st) NDPE% 1 2 Mean 1 2 Mean 1 2 Mean 47É7c2 ^2É30 60É7b ^3É67 69É5a ^2É11 46É2c ^1É04 56É8b ^2É08 66É9a ^1É88 47É0c ^1É80 58É8b ^3É41 68É2a ^2É30 53É0c ^2É94 65É1b ^4É42 77É0a ^2É58 56É2c ^1É45 61É1b ^2É55 73É1a ^2É28 54É6c ^2É72 63É1b ^3É74 75É0a ^3É03 5É6b ^0É27 5É6b ^0É34 6É8a ^0É21 5É1b ^0É16 5É7b ^0É21 6É3a ^0É18 5É3b ^0É79 5É6b ^0É26 6É6a ^0É32 a Five weanling Sprague-Dawley rats per group were individually fed for 10 days. Means within the same letter in the same column are not signiÐcantly di†erent (P \ 0É05). DISCUSSION Proximate and micronutrient analysis The results have indicated di†erence in proximate analysis among black cumin seeds of di†erent sources. Such variation among plant species and even varieties is expected, and it depends on many factors such as the country of origin, stage of maturity, growing and subsequent storage conditions (Holland et al 1992). The proximate analysis data of black cumin seeds have been reported by few authors (Gad et al 1963 ; Babayan et al 1978 ; Ustum et al 1990). It is indicated from their published values that there have been di†erences in the concentrations of fat, moisture, ash, crude Ðbre, protein and carbohydrate. Again, this may be due to geographical and climatic di†erences of areas where the seeds had been grown, or due to the di†erences in the analytical techniques used. The results obtained in the present study show that the fat contents of black cumin are relatively high. Data on protein and micronutrient contents indicate that black cumin is a good source of protein and many vitamins and minerals, whether taken as absolute values or INQ values. Guthrie (1989) considered that a foodstu† is nutritious if four of its nutrients have INQ values exceeding 1É0. In the present investigation, protein, iron, copper, phosphorus, zinc, thiamin, pyridoxine and niacin had INQ values greater than 1É0. Four minerals, namely copper, phosphorus, zinc and iron, were relatively high in black cumin. However, the nutritional status of these minerals cannot be predicted from the apparent quantity ; the quantity of black cumin consumed and the bioavailability of these nutrients in the seeds a†ect the real contribution of black cumin for minerals. It is documented that the absorption and utilisation of minerals in foods of plant origin are inÑuenced by many factors, such as the chemical from the nutrients that occur in food, the level of intake, the presence of binding agents and other nutrients in food ingested simultaneously with the mineral elements, besides other factors associated with the nutrient (Holland et al 1992). Net protein utilisation and protein efficiency ratio experiments The NPU value reÑects nitrogen intake which is retained in the body. The NPU operative values are obtained at actual protein levels of the diet and are a†ected, therefore, by such levels (Pellett and Young 1980). For this reason, the NPU standardised is used to compare the NPU value at maintenance levels of protein (Pellett and Young 1980). However, since the protein contents in this study are similar for all diets, TABLE 9 Weight gain, feed intake and feed conversion ratio for Syrian black cumin in rats fed diets for 28 daysa Run W eight gain (X1 ^ SD) Feed intake (X1 ^ SD) Feed conversion ratio PER Adjusted PERb First Second Casein 45É7 ^ 3É29 44É7 ^ 3É03 67É4 ^ 9É55 209É2 ^ 14É53 211É6 ^ 24É63 247É1 ^ 27É52 4É6 4É7 3É7 1É8 1É8 2É3 2É0 1É9 2É5 a Five weanling Sprague-Dawley rats per group were individually fed for 28 days. b Adjusted PER \ PER of diet ] 2É5 . PER of casein H R H T akruri, M A F Dameh 410 the di†erence in NPU operative did not change substantially when NPU was expressed as NPU standardised. The NPU standardised values for Syrian black cumin diet and Turkish black cumin diet were 54É6 and 63É1, respectively. These values, when compared with other kinds of foods (Williams 1993), suggest that it is possible to rank black cumin in a good nutritional position and consider its protein as having good quality. The di†erence between the NPU values of the Turkish and Syrian black cumin is not easy to explain, since no botanical data are available. However, it is documented that di†erent sources of the same plant may have di†erent protein quality (Abdel-Rahman 1983 ; Mansour et al 1983). It is worth noting that the Ðbre content of the Syrian sample was very high and had a lower NPU value than the Turkish samples. It is possible, therefore, that the high Ðbre adversely a†ected the digestibility and thus the NPU value (Williams 1993). It is also worth noting that Babayan et al (1978) have reported that black cumin seeds have high contents of both methionine and lysine, the two limiting amino acids in legumes and cereal proteins, respectively. From their data, one expects to Ðnd a much higher NPU value. Since this was not the case, one can hypothesise that either another essential amino acid is deÐcient or that there is a di†erence in the amino acid patterns of the variety studied by these authors and the two varieties studied in this work ; probably there is another reason. In any case, this discrepancy awaits explanation through further studies which consider the chemical score and amino acid analysis. NDPE% was used to express both the quality and the quantity of dietary proteins (Miller 1963). As shown in Table 8, NDPE% was 5É3% for the Syrian black cumin diet, 5É6% for the Turkish black cumin diet and 6É6% for the casein diet. Platt et al (1961) recommended an NDPE% value of dietary protein mixtures of at least 5É9 for young children and 4É6 for adults. However, it should be noted here that black cumin seeds are not consumed alone but as a food mixture with other foods (Halva and Lanska 1980). Inspite of this, its protein is expected to have a good complementary value. It is concluded from the overall results of protein quality evaluation, namely those of NPU, PER and NDPE%, together with proximate analysis and vitamins and mineral analyses, that black cumin seeds are a relatively good source of protein and other nutrients and should be evaluated for use in food mixtures for human consumption. However, further investigation, especially on the mineral bioavailability and amino acid score, is needed. REFERENCES Abdel-Rahman A H Y 1983 Amino acid composition of two Italian Durum wheat varieties and their semolina. Food Chem 10 155È158. AOAC 1990 Official Methods of Analysis (15th edn). 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