Int. J. Biosci.

2020

International Journal of Biosciences | IJB |

ISSN: 2220-6655 (Print), 2222-5234 (Online)
http://www.innspub.net
Vol. 16, No. 2, p. 80-88, 2020

RESEARCH PAPER OPEN ACCESS

Nitrate accumulation in the winter and summer vegetables

grown in the Punjab (Pakistan)

Muhammad Mubashir1*, Niaz Ahmed2, Ashfaq Ahmad Rahi8, Sadiq Muhammad3,

Hafiz Saeed-ur-Rehman1, Ghulam Murtaza Khan4, Khurram Shehzad Baig6,
Muhammad Akram7, Munawar Mehdi5

1
Soil and Water Testing Laboratory for Research, Bahawalpur, Punjab, Pakistan
2
Department of Soil Science, Faculty of Agricultural Sciences and Technology, Bahauddin Zakariya
University Multan, Punjab, Pakistan
3
Soil and Water Testing Laboratory for Research, Lahore, Punjab, Pakistan
4
Soil and Water Testing Laboratory for Research, Lodharan, Punjab, Pakistan
5
Directorate of Soil Fertility and Research Institute Lahore, Punjab, Pakistan
6
Soil and Water Testing Laboratory, Muzaffargarh, Punjab, Pakistan
7
Soil and Water Testing Laboratory for Research, Multan, Punjab, Pakistan
8
Pesticide Quality Control Laboratory Multan, Punjab, Pakistan

Key words: Nitrate accumulation, Fertilizer use, Season, Leafy vegetables.

http://dx.doi.org/10.12692/ijb/16.2.80-88 Article published on February 05, 2020

Abstract
Vegetables occupy a pivotal place in the human diet, but unfortunately these contribute maximally to nitrate accumulation and
pose serious health hazards. Nitrite and nitroso compounds which form when nitrite binds to other substances before or after
ingestion e.g. the amines derived from proteins are toxic and can lead to severe pathologies in humans. Thus, the assessment
of the health risk of nitrate to humans should encompass the toxicity of both nitrite and nitroso compounds. That’s why a
comprehensive study was carried out to determine the nitrate accumulation in winter and summer vegetables throughout the
Punjab province of Pakistan. The winter vegetables accumulated more NO3 content than summer vegetables. On the other
hand leafy vegetables accumulated more nitrates than others. Among the winter vegetables radish and hybrid spinach
remained at the top regarding NO3 accumulation. Whereas among the summer vegetables i.e. brinjal and squish secured the
highest position in this regard. The factors affecting the nitrate content viz season (winter and summer), growing intensity of
agriculture fertilizer use and soil texture were also studied.
* Corresponding Author: Muhammad Mubashir  mubashir116@yahoo.com

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Introduction concentration as the crop approaches harvest. Many

Nitrogen fertilization facilitates accumulation of workers have determined the nitrate and nitrite
nitrate in plant tissues as a result of an excess of contents of the prepared infant cereal foods and
nitrogen uptake over its reduction. When taken up in common vegetables. The leafy vegetables contained
excess of immediate requirement, it is stored as free large amount of nitrate and nitrite. The root
nitrate in the vacuole and can be remobilized vegetables like beets, turnips and radish were
subsequently when nitrogen supply is insufficient to reported to have the highest nitrate contents (1000 to
meet the demand (Van der Leij et al., 1998). 2000 ppm). The potato tubers were found to contain
less than 100 ppm nitrates. In tomatoes the
Application of nitrogenous fertilizers has been concentration of nitrates ranged from 25 to 140 ppm.
increased in fields to increase the crop yields. Since Heisler et al. (1974) reported a rapid increase of
Nitrogen fertilizer plays imperative role both in crop nitrite contents in storage abused spinach, beets and
yield and quality (Sisson et al., 1991; Gastal and beet juice. Tamme et al. (2010) observed that the
Lemaire, 2002; Wang et al., 2002). Nitrate is often average levels of nitrate varied from zero to 9669 mg
the major source of Nitrogen available to higher as KNO3 Kg-1 in spinach, beetroots and radish.
plants especially to vegetables. Nitrate uptake and Highest level of nitrates found in leafy vegetables
distribution in crops is of major importance with followed by root vegetables. Cooking had the effect of
respect to both environmental concerns and the lowering the levels of nitrate in all types of fresh
quality of crop products. vegetables. No nitrite was formed during cooking.

Plants have shown a greater tendency for nitrate Approximately 5% of all ingested nitrate is converted
accumulation, especially in vegetables. Because edible in saliva and the gastrointestinal tract to the more
parts contain very high concentrations of nitrate that toxic nitrite (Pannala et al., 2003). The toxic effects of
has been implicated in the occurrence of nitrate are those resulting from the nitrite formed by
methaemoglobinemia and possibly in gastric caner as its reduction by bacterial enzymes (Mensinga et al.,
well as other diseases (Bruning-Fann, 1993; Ishiwata 2003). Nitrite and nitroso compounds which form
et al., 2002). Accumulation of nitrate in high amount when nitrite binds to other substances before or after
in crops is becoming a major concern (Van der Boon ingestion e.g. the amines derived from proteins are
et al., 1990). It creates imbalance between toxic and can lead to severe pathologies in humans.
translocation and intake of nitrates via xylem, and Thus, the assessment of the health risk of nitrate to
conversion of nitrates to ammonia that is rapidly humans should encompass the toxicity of both nitrite
assimilated into amino acids (Maynard et al., 1976). and nitroso compounds. The best effect of nitrite is
Nitrates concentration in lettuce is affected reaction with haemoglobin (oxyHb) which produce
depending upon source of N fertilizer and texture of methaemoglobin (meth Hb) and nitrate.
soil (Gianquinot et al., 1992; Gunes et al., 1995). The
NH4-N-to-NO3-N fertilizer-N ratio (Demsar and
Osvald, 2003), the timing of fertilizer N release (Tesi As a consequence of the formation of methHb the
and Lenzi, 1998), the light intensity and duration oxygen delivery to tissue is impaired (Mensiga et al.,
(Behr and Wiebe, 1992), crop season (Gianquinto et 2003). Generally, nitrate accumulating vegetables
al., 1992), and lettuce type and cultivar. In most types belong to the families of Brassicaceae (radish,
of lettuce, including the romaine type, the highest mustard) Chenopodiaceae (Swiss chard, lettuce and
concentration of nitrates is normally observed in the Apiaceae (celery, parsley) (Santamaria, 2006). In this
external leaves (Abu Rayyan et al., 2004). Plants that study, nitrate contents in winter and summer
develop fruit or storage organs such as tomatoes or vegetables grown in the Punjab province of Pakistan
potatoes usually show a decline in nitrate have been measured.

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Materials and methods determine plant dry weight. Dried plants were ground
Collection and processing of vegetable samples in a mill (40 mesh). Then nitrate (NO3) contents were
A survey of the 36 districts (8 Divisions) of the Punjab determined by method reported by Ryan et al.
province was carried out to procure the summer and (2002). All results are expressed as Means on dry
winter vegetable samples. From each district nine weight basis.
vegetable samples were taken. From four samples
from each side within the municipal limit (urban Statistical analysis
area) and four samples from the rural area (4-5 km) Results were analyzed by analysis of variance using
and one sample was collected from the centre of the the SAS computer program (SAS Institute, 1988).
city center (Fig. 1).
Data was subjected to multivariate statistics such as
The samples of these matured crops were collected Mean and standard Error of Mean, Analysis of
and wrapped in polyethylene bags. The samples of Variance, and Duncan Multiple Range Test (DMRT).
leafy and other vegetable were stored in refrigerator A probability level of P ≤ 0.05 was chosen to establish
at 4°C temperature during transportation and prior to statistical significance.
analysis. This procedure was carried out to avoid the
reduction of nitrate into nitrite during storage. For Results and discussion
leafy vegetables the samples were washed with Season and Nitrate Accumulation
distilled water and in case of fruits it was chopped The data regarding NO3 accumulation in winter
into small portions, dried with blotting paper and vegetables in the eight Divisions of the Punjab
then dried for 48 hours at 700C and weighed to province is presented in Table 1.

Table 1. Nitrate accumulation (mg kg-1) in different winter vegetables in eight divisions of Punjab province of
Pakistan.
Division Carrot Hybrid Spinach Local Spinach Radish Turnip Potato
Rawalpindi 1309c - 1925bc 5273b 904c 787g
Gujranwala 1631a 5535b 1998b 5755ab 955abc 954a
Lahore 1608a 4667f 2106a 6065a 922bc 930c
Faisalabad 1570a 5532c 1991b 5671ab 886c 909d
Sargodha 1570ab 5940a 2001b 5667ab 973abc 885e
Multan 1426bc - 1885c 5604ab 1080ab -
Bahawalpur 1496ab 5050e 1841c 5586ab 807c 795f
Dera Ghazi Khan 1638a 5206d 2022ba 5810ab 1091a -
Average 1531 3991 1971 5679 952 658
DMRT Means with same letter are not significantly different.

On average, the nitrate contents of winter vegetables The data relating to NO3 content in summer
ranged from 657 to 5679 mg kg-1. Results indicated vegetables of the eight divisions of the Punjab
that significant variations among the Divisions in province is presented in Table 2. In general, brinjal
different winter vegetables were existent. The and squash accumulated more nitrate contents than
maximum NO3 contents (5678 and 3991 mg kg-1) onion, okra and tomato. Population quantum exerted
were accumulated in radish and hybrid spinach significant effect on the nitrate accumulation in
respectively. The minimum NO3 contents (952 and different vegetables (Table 3 and 4). The vegetables
657 mg kg-1) were present in turnip and potato collected from large cities accumulated high nitrate
respectively. contents.

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Table 2. Nitrate accumulations (mg kg-1) in different summer vegetables in eight divisions of Punjab province of
Pakistan.
Division Okra Brinjal Squash Onion Tomato
Rawalpindi 998abc 1993ab 1354a 1524a 410a
Gujranwala 981bc 1765d 1448a 949g 352dc
Lahore 1075a 2075a 1954a 1019e 376bc
Faisalabad 992abc 1863dc 1456a 1014f 355dc
Sargodha 1044ab 2018a 1562a 1333b 400ab
Multan 972bc 1886bc 1338a 912h 345d
Bahawalpur 928c 1894bc 1802a 1030d 357dc
Dera Ghazi Khan 952c 1964abc 1618a 1043c 390ab
Average 993 1932 1567 1103 373
DMRT Means with same letter are not significantly different.

Table 3. Effect of population on the Nitrate accumulation (mg kg-1) in different winter vegetables in Punjab
province of Pakistan.
Population Carrot Hybrid Spinach Local Spinach Radish Turnip Potato
Large 1524a 6088a 2109a 5767a 1008a 891a
Medium 1579a 5323b 1960b 5770a 977a 885c
Small 1508a 5321c 1938b 5587a 933a 886b
Average 1537 5577 2002 5708 973 887
DMRT Means with same letter are not significantly different.

The concentration was highest in radish (5708 mg kg- accumulated nitrates as 1943 mg kg-1 and 1566 mg kg-
1) followed by hybrid spinach (5577 mg kg-1). The 1 respectively while tomato accumulated nitrate
minimum concentration was found in potato (887 mg contents (372 mg kg-1 ) followed by Okra (1014 mg kg-1
kg-1) and turnip (927 mg kg-1). While in summer ) and onion (1056 mg kg-1 ).
vegetables (Table 4), on average, brinjal and squash

Table 4. Effect of population on the Nitrate accumulation (mg kg-1) in different summer vegetables in Punjab
province of Pakistan.
Population Okra Brinjal Squash Onion Tomato
Large 1083a 2023a 1554a 1074b 390a
Medium 992b 1900b 1652a 972c 360b
Small 968b 1907b 1492a 1107a 367b
Average 1014 1943 1566 1051 373
DMRT Means with same letter are not significantly different.

Table 5. Effect of sewage water on the Nitrate accumulation (mg kg-1) in different winter vegetables in Punjab
province of Pakistan.
Sewage water use Carrot Hybrid Spinach Local Spinach Radish Turnip Potato
High 1613.0a 5493a 2046.0a 5896.4a 1038.3a 923.8a
Moderate 1475.2b 5297b 1887.7b 5382.0b 899.7b 851.7c
Low 1368.0c 5226c 1870.4b 5543.6b 817.1c 865.0b
Average 1485.4 5338.7 1934.7 5607.3 918.4 880.2
DMRT Means with same letter are not significantly different.

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Use of sewage water and nitrate accumulation the minimum concentration (880.2 mg kg-1 and
The data concerning the impact of sewage water on turnip 918 mg kg-1) in winter vegetables.
nitrate accumulation in vegetables is shown in Tables
5 and 6. The data depicted where the frequency of The similar trend was observed in Summer vegetables
sewage water uses was more the nitrate accumulation such as brinjal (1887 mg kg-1 and squish 14.95 mg kg-
was also high. On average radish (5607 mg kg-1) and 1) accumulated more nitrate then tomato (358 mg kg-1
hybrid spinach accumulated nitrate contents, while and Okra 959.5 mg kg-1 ).

Table 6. Effect of sewage water on the nitrate accumulation (mg kg-1) in different summer vegetables in Punjab
province of Pakistan.
Sewage water use Okra Brinjal Squash Onion Tomato
High 1046.3a 2001.0a 1628.6a 1071a 382.2a
Moderate 946.4b 1817.4b 1529.3a 1038c 363.9b
Low 885.9c 1844.4b 1329.9a 1044b 330.5c
Average 959.5 1887.6 1495.9 1051 358.9
DMRT Means with same letter are not significantly different.

Growing industry and nitrate accumulation accumulated low nitrate compared to potato and
The data showing the impact of growing intensity on turnip growing under the same conditions in winter.
nitrate accumulation is presented in Tables 7 and 8. While in Summer Brinjal and Squash accumulate
Medium growing industry affected nitrate more nitrate contents than Tomato and Okra.
concentration in both the season. Radish and spinach

Table 7. Effect of growing intensity on the nitrate accumulation (mg kg-1) in different winter vegetables in Punjab
province of Pakistan.
Growing Intensity Carrot Hybrid Spinach Local Spinach Radish Turnip Potato
High 1530b 5386b 1964ab 5738a 948b 895b
Medium 1657a 5421a 2014a 5683a 1036a 897a
Low 1315c 5100c 1933b 5356b 892b 832c
Average 1501 5302 1970 5592 959 874
DMRT Means with same letter are not significantly different.

Table 8. Effect of growing intensity on the Nitrate accumulation (mg kg-1) in different summer vegetables in
Punjab province of Pakistan.
Growing Intensity Okra Brinjal Squash Onion Tomato
High 1000ab 1921a 1578a 982c 363b
Medium 955b 1905a 1594a 1157b 379ab
Low 1030a 1948a 1310a 1331a 384a
Average 995 1925 1494 1157 375
DMRT Means with same letter are not significantly different.

Fertilizer use and nitrate accumulation accumulation. The higher the fertilizer use intensity
The intensity of fertilizer use (nitrogenous fertilizer) the more nitrate accumulation was observed in
use exerted pronounced effect on nitrate vegetables in both the seasons. On average, radish

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 Int. J. Biosci. 2020

(5549 mg kg-1) and hybrid Spanish (5314 mg kg-1) carrot (1511 mg kg-1) while in summer vegetables
accumulated nitrate contents. While the minimum Brinjal was at the top (1919 mg kg-1) while tomato
nitrate concentration was observed in the order of remains at the lowest (372 mg kg-1) (Tables 9 and 10).
potato (877 mg kg-1) > turnip > (947 mg kg-1) and

Table 9. Effect of Fertilizer Use on Nitrate accumulation (mg kg-1) of different Winter vegetables in Punjab
Province of Pakistan.
Fertilizer Use Carrot Hybrid Spinach Local Spinach Radish Turnip Potato
High 1547a 5368b 1978a 5775a 971a 896b
Medium 1547a 5473a 1940a 5512ab 929a 909a
Low 1442a 5100c 1984a 5359b 942a 824c
Average 1512 5314 1967 5549 947 876
DMRT Means with same letter are not significantly different.

Table 10. Effect of Fertilizer Use on nitrate accumulation (mg kg-1) of different winter vegetables in Punjab
province of Pakistan.
Fertilizer Use Okra Brinjal Squash Onion Tomato
High 1002a 1926ab 1600a 1026c 366b
Medium 957a 1858b 1489a 1093b 364b
Low 977a 1972a 1345a 1280a 386a
Average 979 1919 1478 1133 372
DMRT Means with same letter are not significantly different.

Soil texture and nitrate accumulation found in potato (898 mg kg-1) and turnip (926 mg kg-
Data regarding the effect of soil texture on nitrate 1).

accumulation in vegetables has been presented in

Tables 11 and 12. It was observed that lighter the During the summer the similar behavior was noticed.
texture more would be the accumulation of nitrate Brinjal (1926 mg kg-1) and squish (1539 mg kg-1)
compared to the heavy texture. Radish (5745 mg kg-1) remained the higher accumulator of nitrate while
and hybrid spinach (5607 mg kg-1) accumulated the tomato (372 mg kg-1), Okra (1011 mg kg-1) and onion
nitrate at the par. While the lowest concentration was (1075 mg kg-1) remained the lower accumulator.

Table 11. Effect of Soil Texture on Nitrate accumulation (mg kg-1) in different winter vegetables of Punjab
province, Pakistan.
Soil Texture Carrot Hybrid Spinach Local Spinach Radish Turnip Potato
Loam 1474b 4976d 1903b 5567a 945ab 871c
Sandy Loam 1689a 5559c 2044a 5844a 1037a 867d
Clay Loam 1487b 5850b 2042a 5775a 930b 915b
Sandy Clay Loam 1589ab 6046a 2054a 5798a 903b 939a
Average 1560 5608 2011 5746 954 898
DMRT Means with same letter are not significantly different.

Table 12. Effect of Soil Texture on Nitrate accumulation (mg kg-1) in different winter vegetables of Punjab
Province, Pakistan.
Soil Texture Okra Brinjal Squash Onion Tomato
Loam 970a 1906bc 1535a 954d 358b
Sandy Loam 1004a 1964ab 1639a 1259a 389a
Clay Loam 1038a 1827c 1513a 1048b 375ab
Sandy Clay Loam 1034a 2008a 1472a 1040c 370ab
Average 1012 1926 1540 1075 373
DMRT Means with same letter are not significantly different.

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The limits set by European Commission (2002) Agriculture 84(9), 931-936.

regarding nitrate accumulation vary according to https://doi.org/10.1002/jsfa.1733
season indicating higher nitrate levels for winter
grown vegetables compared to those grown in the Behr U, Wiebe HJ. 1992. Relation between
summer. In fact, higher nitrate reductase activity was photosynthesis and nitrate content of lettuce
observed at increased light levels (Gaurdeau et al., cultivars. Scientia Horticulturae 49(3-4), 175-179.
1995) and a close negative correlation between https://doi.org/10.1016/0304-4238(92)90155-6
photosynthetic activity and nitrate content has been
reported for various lettuce cultivars (Behr and Blom-Zandstra M, Eenink AH. 1986. Nitrate
Wiebe, 1992). concentration and reduction in different genotypes of
lettuce. Journal of the American Society for
Conclusion Horticultural Science 111(6), 908-911.
The vegetables grown in winter season contained
higher content of NOˉ3 –N than summer season. Bruning-Fann CS, Kaneene JB. 1993. The effects
Vegetables namely, carrot, hybrid spinach, of nitrate, nitrite and N-nitroso compounds on
indigenous (Local) spinach, radish, turnip and potato human health: a review. Veterinary and human
grown in Gujranwala Division contained higher toxicology 35(6), 521-538.
quantities of nitrates compared to other Divisions.
Corre WJ, Breimer I. 1979. Nitrate and nitrite in
Furthermore, vegetables frequently irrigated by vegetables. Centre for Agricultural Publishing and
sewage water contained higher content of nitrates Documentation, Wagheningen, p 85.
compared to moderate irrigated frequency areas. The
accumulation of nitrates by various vegetables was Demsar J, Osvald J. 2003. Influence of NO3-: NH4+
greatly affected by various cropping growing ratio on growth and nitrate accumulation in lettuce
intensity. (Lactuca sativa L. var capitata) in an aeroponic
system. Agrochimica (Italy).
Acknowledgement
This paper is a part of PhD thesis of Mr. Muhammad Gastal F, Lemaire G. 2002. N uptake and
Mubashir. T. M. Ansari, S. A. Malik and A.A. Khan distribution in crops: an agronomical and
acted as PhD Supervisors. Other Co-authors provided ecophysiological perspective. Journal of Experimental
laboratory facilities and were actively involved in this Botany 53(370), 789-99.
study. https://doi.org/10.1093/jexbot/53.370.789

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