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Quantification and detoxification of aflatoxin in food items.

Introduction

Aflatoxins are toxic and carcinogenic metabolites produced by species of Aspergillus, especially Aspergillus flavus and Aspergillus parasiticus. The toxic effects include acute hepatitis, immune-suppression. In humans, the risks associated with aflatoxin consumption are well documented and the International Agency for Research on Cancer (IARC) has designated aflatoxin as a human liver carcinogen. Because of these toxic effects, the Food and Drug Administration regulates the aflatoxin concentration in food with aflatoxin. Commodities or food with aflatoxin exceeding 20 ppb (ug/kg) cannot move in trade (Wogan, 1999). Very little was known about mould metabolites prior to 1961. In that year some alarming reports of a mysterious disease of Turkey poults came from South East of England, tentatively named as Turkey X disease. In 500 such outbreaks about 1, 00,000 poults, mostly between three to six weeks of ages, died. Similarly 5000 partridges and pheasants from one farm and 14000 ducklings from another farm died (Asplin and Carnaghan, 1961). Reports from other places also accumulated on outbreak of acute poisoning of farm animals such as pigs (Loosmore and Harding, 1961) and calves (Loosmore and Markson, 1961). Bioassay test on duckling also helped in furthering and establishing the toxic factor (Sargeant et al., 1961). Aflatoxins producing ability is confined to A. flavus and A. parasiticus. Strains of these two species are common and wide-spread and have been isolated from a number of different host materials. Colonies of A. flavus are green-yellow to yellow-green and that of A. parasiticus are dark green. The toxin is produced by mycelium and secreted into the medium or substrate, spores contain very little aflatoxin. Different strains of A. flavus produce varying amount of aflatoxin and same strains also produce varying amount of aflatoxins. Aflatoxin production is a genetical process depending on specific nutrient and environmental factors (Patterson, 1973). Investigation carried out by various researchers at Tropical Development and Research Institute, London indicated that A. flavus is found in the soil and air throughout the world. Both A. flavus and A. parasiticus are more prevalent in warmer climate and these moulds can be isolated from stores, dried stuff and tropical soil (Christensen, 1957). Aflatoxins naturally occur in rice (including brown, white, black, red and basmati) of different countries, including United State, Canada, Pakistan, India and Thailand (Bansal et al., 2011). Aflatoxins (AFB1, AFB2, AFG1 and AFG2) also occur in freshly harvested corn grains in different regions of Brazil (Liliana et al., 2009). The presence of aflatoxins in red chillis may be a great threat to the health of populations. Total 183 samples of red chilli were screened out for aflatoxin determination. 48 samples were positive for Aflatoxins B1 with the range from 1.2 ppb to 968.3 ppb. Aflatoxin B2 was detected only in 3 samples with the range of 0.3 ppb -159.8 ppb. Aflatoxin G1 and G2 were absent in all chilli samples (Nisa et al., 2012). Chronic poisoning of aflatoxin results in cancer (hepatocellular carcinoma) because liver is the target organ of aflatoxins. Acute intoxication of aflatoxins in human body is also lethal (Milita et al., 2010). Many countries regulate aflatoxin levels in their foods. USA and EU (Europe Union) permit level lower than 20 ppb and Korea and Japan 10 ppb (Chiavaro et al., 2001). Due to its importance different food items were selected for this study and different chemicals were used for detoxification of aflatoxins in these food samples.

Materials and Methods

This study was conducted in Food and Biotechnology Research Centre of PCSIR Laboratories Lahore. The food samples were prepared for aflatoxin analysis (Begum et al., 1985). Aflatoxins were detected by Romers' method (Romer, 1975). Estimation of aflatoxins in toxic extracts was made by comparison with standard technique (AOAC, 2005). In this study, TLC technique was used for the determination of Aflatoxin in all samples.

Nature of samples. Samples of food such as Corn, Wheat, Wheat Flour, White Rice, Brown Rice, Super Basmati, Super Kernal Rice, Red Chilli, Pistachio, Cornflakes, Figs, Haldi, Garam Masala, Peanut, Kalwangi, Makhana and Dates were selected for the present study.

Sample collection. During research work food samples were collected from local market of the city and brought to the Laboratory for quantitative determination and detoxification.

Sampling. Since the aflatoxins are not uniformly distributed in commodities, grains were likely to have pockets of high aflatoxin concentration, firstly due to highly heterogeneous distribution of aflatoxins and secondly due to marketing in lumps of various sizes. To obtain most representative sample, a suitable sampling plan was adopted. These commodities were found stacked in jute bags and stored in house type godown. In order to obtain a more representative portion of these samples, 500 g were collected through a sample probe directly in plastic bags piercing jute bags diagonally from 2 to 3 places. They were passed through sample divider and reduced to approximately 200 g for the purpose of analysis and thus a greater homogeneity of contaminated portion was achieved. Each sample was then thoroughly mixed, ground and made into fine powder for experimental analysis.

Extraction. Extraction procedures and analytical methods vary from one commodity to another because of diverse chemical composition, preventing the development of any one method which could be applied uniformly to all products. However, extraction with chloroform is most suitable method for aflatoxins ([B.sub.1], [B.sub.2], [G.sub.1], [G.sub.2], [M.sub.1] and [M.sub.2]).

Then test portion was taken from mixture. 50 g of ground sample was kept into 500 mL conical flask and 25 mL water and 150 mL chloroform was added into flask. Conical flask was shaken on wrist action shaker for 30 min and sample was filtered through filter paper. 50 mL chloroform was taken into beaker and put on steam bath for evaporation.

Chromatographic tank. The dilutions for spotting were got in micro liter. The 25 [micro]L spot of test solution was applied on thin layer chromatography plate with micro syringe. Spot of 5 or 10 [micro]L of aflatoxins ([B.sub.1], [B.sub.2], [G.sub.1], [G.sub.2], [M.sub.1] and [M.sub.2]) standard was spotted on same plate as an internal standard. The plate was developed with anhydrous ether in thin layer chromatographic tank upto half then removed and dried. Then plate was redeveloped in the same direction in thin layer chromatographic tank with acetone-chloroform (1:9). Plate was removed and test solution spot was observed for presence or absence of aflatoxins under UV light. If preliminary plate would show that new concentration of test solution required then new concentration were prepared for spotting. Different 1 to 25 [micro]L spots of test solution (3.5, 10.5, 24.9 [micro]L) were spotted on new thin layer chromatographic plate and on the same plate 1 to 25 [micro]L aflatoxins standard was spotted (Braicu et al., 2008).

Detection and Estimation. Fluorescing intensities of sample spots were compared with those of standard aflatoxin spots. In case, fluorescing spot of sample lied between the standard spots, the average value of two standard spots was taken into consideration.

Confirmation. Another very important step in the aflatoxins analysis was the fluorescing sample spots. This was carried out by spraying, evenly the thin layer chromatographic plate with aqueous sulphuric acid (50/50 v/v). After the spraying, thin layer chromatographic plate was allowed to dry and then viewed under UVlight (365nm).

Calculation. Concentration of aflatoxins ([micro]g/kg) present in sample was calculated as follows.

Aflatoxins ([micro]g/kg)= S x Y x V / Z x W

Where:

S = volume in [micro]L of aflatoxins standard of equivalent intensity to Z ([micro]L of sample)

Y = concentration of aflatoxins standard in [micro]g/ mL

V = volume in [micro]L of solvents required to dilute final extract

Z = volume in [micro]L of sample extract required to give fluorescence intensity comparable to that of S = [micro]L of aflatoxins standard

W = weight in g of original sample contained in final extract

Treatment for detoxification. Fifty grams of grinded samples in which aflatoxins had been detected were kept in separate 500 mL conical flasks. Chemical solutions of 0.1% HCl, 0.3% HCl, 0.5% HCl, 10% citric acid, 30% citric acid, 05% calcium hydroxide, 0.2 and 0.3% NaOCl, 96% ethanol and 99% acetone were added into different flasks (Table 1). Conical flasks were shaken on wrist action shaker for 2 h and sample was filtered through filter paper and dried for two days.

Quantification after detoxification. Quantification of detoxified sample for aflatoxins was carried out by same method such as chloroform extraction, detection by thin layer chromatography, estimation through UV light and calculation by formula.

Statistical analysis. The statistical significance of the data was analyzed (p=0.042) using pair t-test (Steel et al., 1997).

Results and Discussion

Aflatoxins were detoxified by the treatment of different chemical solutions. For this purpose, total 30 samples were collected. These 30 samples were quantified using thin layer chromatography (TLC) for the presence of aflatoxins level in food items (Table 2). The aflatoxins were not found in 10 samples of food product, remaining 20 samples of food in which aflatoxins had been found were treated with chemical solutions.

Chemical solutions 0.1%, 0.3% and 0.5% of hydrochloric acidreduced aflatoxins to 39.7%, 55.1%, 90.9%, 39.5%, 62.0% and 82.0% in food items (Table 4) which are in line with work of Aly and Hathout (2011) who reduced aflatoxins 27.6%, 42.5% and 90% in food with concentrations of hydrochloric acid at different hours. Aflatoxins also reduced to 49.3%, 86.5% and 71.39% with concentration of 0.1%, 0.3% and 0.5% of hydrochloric acid which is same work as Aly and Hathout (2011) who did reduction of food items.

Aflatoxins reduced to 31.3%, 64.3%, 19.8%, 28.08%, 70.5% and 83.05% with treatment of 10% citric acid, 30% citric acid, 1% sodium bisulphate, 2% sodium bisulphate, 0.2% sodium hypochlorite and 0.3% sodium hypochlorite in food items and Aflatoxins reduced to 63.0%, 70.0%, 69.16%, 53.9%, 10.0% and 35.05% with treatment of 10% citric acid, 30 % citric acid, 1% sodium bisulphate, 2% sodium bisulphate 0.2% sodium hypochlorite and 0.3% sodium Hypochlorite in food items which are in line with work of Mukendi et al. (1991). They had detoxified aflatoxins by comparing chemicals citric acid, sodium bisulphate, sodium hypochlorite (Table 3 and Table 4).

The present study showed significant detoxification in aflatoxins (p = 0.042) of food items when pair T-test was applied to quantified aflatoxins of food items before and after detoxification. It means that aflatoxins had been reduced statistically when aflatoxins were compared before and after detoxification.

Present study also showed that thin layer chromatography is a reliable method for detection and quantification of aflatoxins in food items before and after detoxification. (Okwu et al., 2010, Olufunmilayo and Oyefolu, 2010). Ultra-violet (UV) light, a standard procedure was used to differentiate the toxin from non-toxin forms of A. spergillus species.

Conclusion

It was concluded that significant detoxification i.e. 90.9% was observed when 0.5% HCl was used as detoxifying agent for aflatoxin B1 in Super Basmati rice. Similarly, 83.05% detoxification of Aflatoxin B1 was observed in Dal Mash, 82.09% in Haldi, 82% in Pistachio and 80% in White rice when 0.3% NaOCl, 50% Ca[(OH).sub.2], 0.5% HCl and 5% Ca[(OH).sub.2] were used, respectively.

References

Braicu, C.,Puia, C., Bodoki, E., Socaciu, C. 2008. Screening and quantification of aflatoxins and ochratoxin A in different cereals cultivated in Romania using thin layer chromatography. Journal ofFoodQuality, 31: 108-120.

Chiavaro, E., Dall., A.Sta, C., Galaverna, G., Biancardi, A., Gambarelli, E., Dossena, A., Marchelli, R. 2001. New reversed-phase liquid chromatographic method to detect aflatoxins in food and feed with cyclodextrins as fluorescence enhancers added to the eluent. Journal of Chromatography A, 937: 31-40.

Christensen, C.M. 1957. Deterioration of stored grains by fungi. TheBotanicalReview, 23: 108-134.

Liliana, O.R., Viviane, K.N., Raquel, B., Tatiana, A.R., Estela, K., Benedito, C. 2009. Mycoflora and co-occurrence of fumonisins and aflatoxins in freshly harvested corn in different regions of Brazil. International Journal of Molecular Sciences, 10: 5090-5103.

Loosmore, R.M., Harding, J.D.J. 1961. Atoxic factor in Brazilian groundnut meal causing liver damage inpigs. Veterinary Record, 73: 1362-1364.

Loosmore, R.M., Markson, L.M. 1961. Poisioning of cattle by brazilian groundnut meal. Veterinary Record, 73: 813-814.

Milita, N.M. Mihaescu, G., Chifiriuc, C. 2010. Aflatoxins--health risk factors. Bacteriol Viruses ParazitolEpidemiol, 55: 19-24.

Mukendi, N., Rollmann, B., de Meester, C. 1991. Detoxification of aflatoxin B1 by different chemical methods and evaluation of the effectiveness of the treatments applied. Journal de Pharmacie de Belgique, 46: 182-188.

Nisa, A., Zahra, N., Firdous, S., Ejaz, N., Hina, S. 2012. Detection of Aflatoxins in various samples of red chilli. Pakistan Journal of Scientific and Industrial Research, SerB: Biological Sciences, 55: 27-29.

Okwu, G.I., Achar, P.N., Sharma, S.K. 2010. Quantification of aflatoxin B1 in ready-to-use food thickeners in South-east geo-political zone in Nigeria. African Journal of Microbiology Research, 4: 1788-1793.

Olufunmilayo, G.O., Oyefolu, A.B. 2010. Natural occurrence of aflatoxin residues in fresh and sun dried meat in Nigeria, The Pan African Medical Journal, 7: 14.

Patterson, D.S.P. 1973. Metabolism as a factor in determining the toxic action of aflatoxins in different animal species. Food and Cosmetics Toxicology, 11: 287-294.

Romer, T.R. 1975. Screening method for detection of aflatoxins in mixed feed and other agriculture commodities. Journal of the Association of Official Analytical Chemists, 59: 500-506.

Sargeant, K., Sheridan, A., Kelley, J.O., Carnaghan, R.B.A. 1961. Toxic products with certain samples of groundnuts. Nature, 192: 1096-1097.

Steel, R.D., Torrie, J.H., Dickey, D. 1997. Principles and Procedures of Statistic: A Biometrical Approach, 3rd edition, McGraw Hills Book Co. Inc, New York, USA.

Wogan, G.N. 1999. Aflatoxin as a human carcinogen. Hepatology, 30: 573-575.

Alim-un-Nisa (a) *, Naseem Zahra (b), Sajila Hina (a), Rizwan Hayat (c) and Nusrat Ejaz (a)

(a) Food and Biotechnology Research Centre, PCSIR Laboratories Complex, Lahore-54600, Pakistan

(b) Pakistan Institute of Technology for Minerals & Advanced Engineering Materials (PITMAEM), PCSIR Laboratories Complex, Ferozepur Road, Lahore-54600, Pakistan

(c) Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore, Pakistan

(received August 1, 2012; revised December 23, 2012; accepted January 24, 2013)

* Author for correspondence; E-mail: nisaalim64@yahoo.com
Table 1. Solutions for detoxification of Aflatoxins in
food items

Food product         Chemical solution
                     for detoxification

Red chilli           0.1 % HCl
                     0.3% HCl
Red chilli           5 % NaOH
Super Sella rice     0.3 % HCl
Super Basmati rice   0.5 % HCl
White rice           5% Ca[(OH).sub.2]
Maize grain          10%Citricacid
Wheat damage         30 % Citric acid
Peanut               99 % Acetone
Figs                 96 % Ethanol
Dates                1 % Sodium bisulphate
Brown rice           2 % Sodium bisulphate
Makhana              5 % KOH
Dal chana            0.2 % NaOCl
Dal mash             0.3 % NaOCl
Corn flakes          0.1 % HCl
Kheer mix            0.3 % HCl
Pistachio            0.5 % HCl
Haldi                5 % Ca[(OH).sub.2]
Kalwangi             10%Citricacid
Dal masoor           99 % Acetone

Table 2. Detection and estimation of Aflatoxins in food products

Sample   Food                 Aflatoxin   S     Y      V        Z
No.      product

1.       Red chilli           [B.sub.1]   0.5   2.02   0.5      4.7
                                                       (1000)
2.       Red chilli           [B.sub.1]   2     2.02   0.51     4.9
                                                       (1000)
3.       Super Sella          [B.sub.1]   2     2.02   0.51     2.9
         rice                                          (1000)
4.       Super Basmati        [B.sub.1]   5     2.02   0.99     0.9
         rice                                          (1000)
5.       Dal mash             Absent      --     --      --      --
6.       White rice           [B.sub.1]   8.5   2.02   0.51     24.9
                                                       (1000)
7.       Pistachio            Absent      --     --      --      --
8.       Haldi                Absent      --     --      --      --
9.       Maize grain          [G.sub.1]   0.5   2.03   0.51     0.9
                                                       (1000)
10.      Wheat damage         [B.sub.1]   0.9   2.02   1470     0.9
11.      Peanut               [B.sub.1]   8.5   2.02   0.51     4.9
                                                       (1000)
12.      Figs                 [B.sub.1]   1.0   2.02   0.50     1.0
                                                       (1000)
13.      Dates                [B.sub.1]   0.5   2.02   1980     3.9
14.      Brown rice           [B.sub.1]   2     2.02   0.51     1.0
                                                       (1000)
15.      Makhana              [B.sub.1]   0.5   2.02   0.51     14.9
                                                       (1000)
16.      Dal chana            [B.sub.1]   0.5   2.02   0.99     14.9
                                                       (1000)
17.      Figs                 Absent      --     --     --       --
18.      Peanut               Absent      --     --     --       --
19.      Dal mash             [B.sub.1]   0.5   2.02   0.5      4.7
                                                       (1000)
20.      Corn flakes          [B.sub.1]   1.0   2.02   1        9.5
                                                       (1000)
21.      White rice           Absent      --     --      --      --
22.      Kheer mix            [B.sub.1]   0.9   2.02   0.51     4.9
                                                       (1000)
23.      Pistachio            [B.sub.1]   1.0   2.02   3.0      2.9
                                                       (1000)
24.      Haldi                [B.sub.1]   1     2.02   1980     2.9
25.      Peanut               Absent      --     --     --       --
26.      Brown rice           Absent      --     --     --       --
27.      Kalwangi             [B.sub.1]   4.0   2.02   0.51     5.0
                                                       (1000)
28.      Super Sella rice     Absent      --    --     --       --
29.      Super Basmati rice   Absent      --    --     --       --
30.      Dal masoor           [B.sub.1]   2.0   2.02   0.91     2.9
                                                       (1000)

Sample   Food                 W       ppb
No.      product

1.       Red chilli           15.04   7.14

2.       Red chilli           16.72   25.15

3.       Super Sella          16.78   42.
         rice
4.       Super Basmati        16.92   656.9
         rice
5.       Dal mash             --      --
6.       White rice           16.70   125.53

7.       Pistachio            --      --
8.       Haldi                --      --
9.       Maize grain          16.65   34.48

10.      Wheat damage         16.67   174.32
11.      Peanut               16.70   21.08

12.      Figs                 16.77   14.9

13.      Dates                16.68   30.76
14.      Brown rice           16.76   123.58

15.      Makhana              16.73   2.09

16.      Dal chana            16.72   8.02

17.      Figs                 --      --
18.      Peanut               --      --
19.      Dal mash             15.04   7.14

20.      Corn flakes          16.27   13.0

21.      White rice           --      --
22.      Kheer mix            16.70   11.32

23.      Pistachio            16.51   167.17

24.      Haldi                16.68   82.68
25.      Peanut               --      --
26.      Brown rice           --      --
27.      Kalwangi             16.72   49.2

28.      Super Sella rice     --      --
29.      Super Basmati rice   --      --
30.      Dal masoor           16.67   76.05

Table 3. Detection and estimation in Aflatoxins in food products
after detoxification

Sample   Food                 Aflatoxin   S     Y      V        Z
No.      product

1.       Red chilli           [B.sub.1]   0.9   2.02   0.99     24.9
                                                       (1000)
2.       Red chilli           [B.sub.1]   0.9   2.02   0.51     4.9
                                                       (1000)
3.       Super Sella rice     [B.sub.1]   0.5   2.02   0.91     2.9
                                                       (1000)
4.       Super Basmati rice   [B.sub.1]   2.5   2.02   1975     2.0
                                                       (1000)
6.       White rice           [B.sub.1]   2.0   2.02   0.51     4.9
9.       Maize grain          [G.sub.1]   9.5   2.03   0.51     24.9
                                                       (1000)
10.      Wheat damage         [B.sub.1]   1     2.02   0.515    1
                                                       (1000)
11.      Peanut               [B.sub.1]   1.5   2.02   0.99     24.9
                                                       (1000)
12.      Figs                 [B.sub.2]   0.5   0.5    0.48     0.9
                                                       (1000)
13.      Dates                [B.sub.1]   2     2.02   0.50     4.9
                                                       (1000)
14.      Brown rice           [B.sub.1]   8.5   2.02   0.51     5.9
                                                       (1000)
15.      Makhana              [B.sub.1]   0.5   2.02   0.51     5.9
                                                       (1000)
16.      Dal chana            [B.sub.1]   1     2.02   487      24.9
19.      Dal mash             [B.sub.1]   0.5   2.02   0.50     24.9
                                                       (1000)
20.      Corn flakes          [B.sub.1]   0.5   2.02   0.51     3.9
                                                       (1000)
22.      Kheer mix            [B.sub.1]   0.9   2.02   0.99     24.9
                                                       (1000)
23.      Pistachio            [B.sub.1]   0.5   2.02   1980     3.9
24.      Haldi                [B.sub.1]   6.0   2.02   0.51     24.9
                                                       (1000)
27.      Kalwangi             [B.sub.1]   7.5   2.02   0.51     14.9
                                                       (1000)
30.      Dal masoor           [B.sub.1]   3.0   2.02   0.50     4.7
                                                       (1000)

Sample   Food                 W       ppb
No.      product

1.       Red chilli           16.67   4.3

2.       Red chilli           16.71   11.34

3.       Super Sella rice     16.67   19.01

4.       Super Basmati rice   16.7    59.68

6.       White rice           16.72   25.51
9.       Maize grain          16.71   23.67

10.      Wheat damage         16.69   62.31

11.      Peanut               16.77   7.2

12.      Figs                 16.56   8.54

13.      Dates                16.72   24.66

14.      Brown rice           16.71   88.87

15.      Makhana              16.72   1.94

16.      Dal chana            16.68   2.36
19.      Dal mash             16.69   1.21

20.      Corn flakes          16.69   7.9

22.      Kheer mix            16.77   4.3

23.      Pistachio            16.68   30.7
24.      Haldi                16.69   14.8

27.      Kalwangi             16.72   31

30.      Dal masoor           15.9    40.9

Table 4. Comparison of Aflatoxins estimation of food products before
and after detoxification

Sample   Food                 Aflatoxin    Estimation
No.      product                           before
                                           detoxification
                                           (ppb)

1.       Red chilli           [B..sub.1]   7.14
2.       Red chilli           [B..sub.1]   25.15
3.       Super Sella rice     [B..sub.1]   42.34
4.       Super Basmati rice   [B..sub.1]   656.9
6.       White rice           [B..sub.1]   125.53
9.       Maize grain          [G..sub.1]   34.48
10.      Wheat damage         [B..sub.1]   174.32
11.      Peanut               [B..sub.1]   21.08
12.      Figs                 [B..sub.2]   14.9
13.      Dates                [B..sub.1]   30.76
14.      Brown rice           [B..sub.1]   123.58
15.      Makhana              [B..sub.1]   2.09
16.      Dal chana            [B..sub.1]   8.02
19.      Dal mash             [B..sub.1]   7.14
20.      Corn flakes          [B..sub.1]   13.0
22.      Kheer mix            [B..sub.1]   11.32
23.      Pistachio            [B..sub.1]   167.17
24.      Haldi                [B..sub.1]   82.68
27.      Kalwangi             [B..sub.1]   49.2
30.      Dal masoor           [B..sub.1]   76.05

Sample   Food                 Chemical                ppb
No.      product              solution for            after
                              detoxification          detoxification

1.       Red chilli           0.1 % HCl               4.3
2.       Red chilli           5% NaOH                 11.34
3.       Super Sella rice     0.3% HCl                19.01
4.       Super Basmati rice   0.5% HCl                59.68
6.       White rice           5% Ca(OH)               25.51
9.       Maize grain          10% Citric acid         23.67
10.      Wheat damage         30% Citric acid         62.31
11.      Peanut               99% Acetone             7.2
12.      Figs                 96% Ethanol             8.54
13.      Dates                1% Sodium bisulphate    24.66
14.      Brown rice           2 % Sodium bisulphate   88.87
15.      Makhana              5% KOH                  1.94
16.      Dal chana            0.2% NaOCl              2.36
19.      Dal mash             0.3%NaOCl               1.21
20.      Corn flakes          0.1%HCl                 7.9
22.      Kheer mix            0.3% HCl                4.3
23.      Pistachio            0.5% HCl                30.7
24.      Haldi                50% Ca[(OH).sub.2]      14.8
27.      Kalwangi             10% Citric acid         31.0
30.      Dal masoor           99% Acetone             40.9

Sample   Food                 Reduction
No.      product              in % age

1.       Red chilli           39.7
2.       Red chilli           54.9
3.       Super Sella rice     55.1
4.       Super Basmati rice   90.9
6.       White rice           80.0
9.       Maize grain          31.3
10.      Wheat damage         64.3
11.      Peanut               63.67
12.      Figs                 42.7
13.      Dates                19.8
14.      Brown rice           28.08
15.      Makhana              7.1
16.      Dal chana            70.5
19.      Dal mash             83.05
20.      Corn flakes          39.5
22.      Kheer mix            62.0
23.      Pistachio            82.0
24.      Haldi                82.09
27.      Kalwangi             36.9
30.      Dal masoor           46.2
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Author:Alim-un-Nisa; Zahra, Naseem; Hina, Sajila; Hayat, Rizwan; Ejaz, Nusrat
Publication:Pakistan Journal of Scientific and Industrial Research Series B: Biological Sciences
Article Type:Report
Geographic Code:9PAKI
Date:Jul 1, 2013
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