INTRODUCTION Nitrates and nitrites occur naturally in the environment and commonly used as food preservatives (Gangolli et al.,1994).

For plant, nitrogen was taken up in the form of nitrate ions (Walters, 1996). Vegetables are estimated to contribute approximately 80-92% and 1643% of the average daily dietary intake of nitrate and nitrite, respectively (Chou et al., 2003). The level of nitrates varies according to the type of vegetables, nitrogen fertilization, temperature and amount of sunlight and other factors. Leafy vegetables accumulate the most nitrates compared to other types of vegetable (Chung et al., 2011; Santamaria et al., 1999). However, nitrite contents are usually extremely low in most fresh leafy vegetables (Chung et al., 2011; Matallana Gonzalez et al., 2010; Chung et al., 2003). Studies have shown that prolonged or adverse storage condition of vegetables particularly nitrate rich vegetables might lead to reduction of nitrate to nitrite (Chung et al., 2004; Philips, 1968) but it varied widely between samples(Schuphan, 1965). However, Chung et al. (2004) were reported that nitrate and nitrite contents in leafy vegetables did not change under refrigerated storage for 7 days. OBJECTIVES Thus, the objectives of the study are: 1. To determine nitrate and nitrite contents 2. To determine physical and chemical quality characteristics of leafy vegetables produced under conventional and organic production systems during 9 days of refrigerated storage

LITERATURE REVIEW Nitrate is a form of nitrogen which occurs naturally and essential in the nitrogen cycle in the environment. Nitrate is formed from fertilizers, decaying plants, manure and other organic residues. Usage of nitrate and nitrite includes as food additive, mainly as preservative and antimicrobial agent. Excessive usage of synthetic nitrogen fertilizers and livestock manure in agriculture may lead to higher content of nitrate in vegetables and drinking water. (Santamaria, 2006). The presence of nitrate in vegetables, water and other food poses threats to human health. Nitrate itself is non-toxic , but the compound induced from nitrate which is nitrite is able to cause methaemoglobinaemia. (EFSA, 2008) Nitrite may also react with amines to form carcinogenic nitrosoamines in the stomach (WHO, 2003). However, nitrate has beneficial effects to human health. Its metabolite, nitric oxide, is associated with signal transduction and cardiovascular protection in human (Du et al., 2007). In plant, Nitrate is mainly to be found in cell vacuoles and is transported in xylem. However, Santamaria et al. (1999) reported that nitrate differed in the various portions of vegetables. Plants depend on nitrate uptake to synthesis amino acid and protein. However, the reduction of nitrate is a process requires energy provided by photosynthesis. Therefore when plants are grown in lowlight conditions, plants tend to have higher nitrate content as the nitrate is stored and not reduced to form amino acids (Gilchrist, 2010). Angolli et al., (1994), the nitrate and nitrite contents of vegetable grown under protected conditions is usually considerably higher than in outdoor vegetables. Besides that, Nitrogen fertilization also has been identified as the major factor that influence nitrate content in vegetables. Whereas the nitrite level on most fresh vegetable are low, but increase dramatically via microbiological reduction of nitrate in vegetables (Santamaria, 2006). On the other hand, nitrate concentration may decrease during storage at ambient temperature (Chung et al., 2004; Philips, 1968).

MATERIAL AND METHOD Two kg fresh leafy choy sum (Brassica rapa var. parachinensis) were obtained at random from the Selangor Wholesale Market and Commercial farming respectively. All vegetables were transported in air-tight, polystyrene iced- boxes to the Postharvest Laboratory and were sorted and graded for uniformity in size. After that, vegetables were washed under tap water to remove any soil and dust particles. Then, the vegetables were rinsed with cold distilled water and dried with paper towels. Each perforated plastic bag was put in a household refrigerator for nine days storage at 5 + 1c. Relative humidity in the refrigerator fluctuated in the range of 40%-60%. Both physical and chemical quality characteristics of vegetables were evaluated every three days: a) Weight loss: was measured by using electronic weighing scale every 3 days as a percentage of vegetable weight at harvest. b) Firmness: was measured using an Instron with a 6mm diameter cylindrical probe at a speed of 20mm/min. c) Leaf Colour: was measured with a chromameter (Minolta CR-300),with a measuring head with an 8mm-diameter measuring area to get colour values of L*, C*, h d) Soluble solids concentration (SSC): was measured by dropping 1-2 drops of filtrate on the glass prism of a hand refractometer (Model N-1E, Atago, Japan). e) Titratable acidity (TA): was determined using the titration method. f) Ascorbic acid (AA): Vitamin C or ascorbic acid (AA) was determined using reversed-phase HPLC to checked the existent of ascorbic acid compound on plant extract after 12 hours storage g) Nitrate and nitrite concentration Nitrate was determined by using QuikChem method. Nitrite content was determined by using diazotizing with sulfanilamide followed by coupling with N-1naphthyl-ethylenediamine dihydrochloride. Experimental design and statistical analysis The design of the experiment was a completely randomised design (CRD) with three replications. Data obtained were subjected to analysis of Regression using OriginPro 8 (OriginLab). A correlation analysis by means of Persons correlation matrix was performed to establish the association between variables RESULT & DISCUSION a) Weight loss: There were significant quadratic relationships between weight loss and storage duration of choy sum produced under conventional system and organically system. According to data, organically produced choy sum had a greater weight loss than conventionally produced choy sum storage time progressed where organically produced choy sum lost 5% of their weight after three days of storage whereas conventionally produced choy sum lost 5% weight after 9 days. These happen probably because of water loss and food reserve loss as result of biochemical activities ( Santi et al., 1992). b) Firmness

There was a significant linear relationship between weight loss and storage duration of choy sum produced under organic system. A contrary result was obtained for conventionally produced choy sum. Choy sum produced under organic system had a inner decrease in firmness as storage time progressed. Prolonged storage duration caused a progressive decrease in firmness of choy sum. Sams (1999) reported that water content of plant has a direct influence on cell turgor, and the degree of cellular hydration resulted in noticeable changes in plant texture. It is generally accepted that minor changes in percentage of water loss can result in undesirable texture changes. In the present study, harvested stages of choy sum produced by conventional and organic production systems were uncertain. Sam,(1999) summarised that texture of vegetables is influenced by environmental, cultural, physiological and genetic factors. Production practices such as fertilization, growth regulator use, irrigation, time of harvest and cultivar selection have a major impact on texture.

c) Leaf colour There were significant quadratic relationships between L* and storage duration of choy sum produced under conventional system. However, there was no relationship between L* and storage duration of organically produced choy sum. There were significant quadratic and linear relationships between C* and storage duration of choy sum produced under both production systems. There were significant quadratic relationships between ho and storage duration of choy sum produced under both production systems. Leaf colour of conventionally grown choy sum had turned darker as storage time prolonged. The decreasing in L*for choy sum produced under conventional system as storage duration prolonged Choy sum produced under conventional system had a quadratic increase in C* as the storage duration prolonged. This showed that 67% of the variability in C* was due to the storage duration. Both leaf colour became more intense with the progressive increase in C* during storage. Leaf colour of choy sum in both production systems turned from green to yellowish green. Thus, the quality of choy sum decreased significantly in terms of leaf colour during storage. The green colour of leafy vegetables is related to chlorophyll content. Change in leaf colour from green to yellowish green indicates onset of chlorophyll degradation (Meir et al., 1992). Chlorophyll degradation has been shown to be affected by many factors such as temperature and relative humidity (Roura et al., 2000; Perrin and Gaye, 1986). Also, chlorophyll loss is a good indication of green tissue senescence, since chloroplast pigments, lipids and proteins are normally the first compounds to be degraded (Pogson and Morris, 2004; Nooden et al., 1997). d) Soluble solids concentration (SSC) There were no significant relationships between SSC and storage duration of choy sum. SSC was not significantly affected by prolonged storage duration. The energy sources available in the tissues of freshly harvested vegetables are free amino acid and carbohydrates (Heimdal et al., 1995). Therefore, sugar consumption during respiration will lead to decrease in SSC. Watada et al. (1996) reported that higher storage temperatures result in higher respiration rates in fresh cut vegetables. In the present study, SSC in choy sum was not affected during storage. Subsequently, this may indicate that respiration rate in choy sum was low during storage at 5c+1c. e) Titratable acidity (TA) There were significant quadratic relationships between TA and storage duration of choy sum produced under conventional system. Contrary results were obtained for organically produced choy sum. TA of conventionally produced choy sum changed in a positive quadratic pattern during storage. Organic acids have a pronounced impact on food flavour and quality (Sadler and Murphy, 2010). TA and SSC are often evaluated in analysis in produce because they are critical components to overall sensory quality. Therefore, decreases in TA and SSC may indicate some degree of reduction in flavour (Mattheis and Fellman, 1999). In the present study, SSC and TA of organically produced choy sum were not affected by storage duration. Therefore, it can be concluded that organically produced choy sum had better retention of flavour than conventionally produced choy sum during storage. f) Ascorbic Acid (AA)

There were significant quadratic relationships between AA content and storage duration of choy sum produced under conventional system. However, opposing results were obtained for organically produced choy sum. Choy sum produced under conventional system had a quadratic decrease in AA content as storage duration prolonged. This indicated that 69% of the variability in AA content was due to the storage duration. AA content decreased after four days of storage. In contrast, AA content in organically produced choy sum was not affected by prolonged storage duration. Hence, retention of AA content was higher in organically produced choy sum than conventionally produced choy sum. They might be because of the usage of agricultural chemicals in conventional production system may affect the lability of ascorbic acid during storage. Agrochemicals such as pesticides and growth regulator may indirectly affect the nutritional quality of vegetables (Moreira et al. 2003). Nevertheless, the differences could be attributed to better harvest and post-harvest practices in organic production system. Proper cold chain system plays a pivotal role in ensuring quality of vegetables especially ascorbic acid. Ascorbic acid is very labile and sensitive to heat, oxygen and light (Eitenmiller et al., 2008). g) Nitrate and Nitrite content There were no significant relationships between nitrate and nitrite content and storage duration of choy sum produced under conventional system. Similar results were obtained for organically produced choy sum. The conversion of nitrate to nitrite in fresh spinach was delayed under refrigerated storage (Philips, 1968). Nitrite did not accumulate under refrigerated storage. Hence, it is practically safe to store leafy vegetables under refrigerated temperature. However, initial nitrate and nitrite contents were considerably high in both conventionally and organically produced choy sum. The nitrate (NO3-) and nitrite (NO2-) contents in conventionally produced choy sum were 107, 850 mg kg-1 and 12.52 mg kg-1 (in dry weight), respectively. Nitrate and nitrite contents in organically produced choy sum were 106,670 mg kg-1 and 13.42 mg kg-1 (in dry weight), respectively. This finding of this study was oppose to William (2002), who reported that organically fertilised crops, particularly leafy vegetables, had lower nitrate content. In addition, Herencia et al. (2011) reported that crops from organic plots had lower nitrate content than those with mineral fertilisation. However, there was high data variability because the presence of factors that could have affected the nitrate content of crop such as cultivar, soil type, planting and harvest dates, nitrate in irrigation water and groundwater, geographical location, climate, storage conditions and plant disease.

CONCLUSION This study verified that nitrate and nitrite contents did not change in fresh, clean and perforated PE bag packed choy sum under refrigerated storage. Hence, it is reasonably safe to keep fresh, clean and perforated PE bag packed leafy vegetables in a refrigerator. However, initial nitrate content in choy sum was unusually high for both conventionally and organically produced choy sum. Ascorbic acid content in organically produced choy sum was not affected by storage duration. However, ascorbic acid content in conventionally produced choy sum decreased significantly after 4 days of storage. Hence, retention of ascorbic acid content was higher in

organically produced choy sum than conventionally produced choy sum. Weight loss and firmness of organically produced choy sum was 102.8 % higher and 54.8% lower, respectively, than conventionally produced choy sum after 9 days of storage. Firmness of conventionally produced choy sum was not affected by storage duration. Leaf colours for both choy sum produced under conventional and organic systems changed from green to yellowish green as storage time progressed. Leaf L* of organically produced choy sum was not affected by storage duration. However, L* of of conventionally produced choy sum showed a negative quadratic pattern during storage. There was visible and drastic change in leaf colour of conventionally produced choy sum during storage. However, there was minimal change in leaf colour of organically produced choy sum. SSC remained unchanged for choy sum in both the production systems during storage. Titratable acidity showed a positive quadratic relationship with storage duration for conventionally produced choy sum. However, titratable acidity in organically produced choy sum was not affected by storage duration. Therefore, it can be concluded that organically produced choy sum had better retention of flavour than conventionally produced choy sum during storage. It is recommended organically produced choy sum be consumed within 3 days of storage, before it loses its fresh appearance and get wilted. Conventionally produced choy sum should be consumed within 4 days as ascorbic acid decreased drastically after 4 days of storage.

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