International Journal of Food Science and Technology 2001, 36, 283289

283

Inuence of oxygen concentration and temperature on respiratory characteristics of fresh-cut green onion
Seok-In Hong* & Dong-Man Kim
Korea Food Research Institute, Bundang, Seongnam, Kyonggi 463-420, Korea (Received 23 January 2000; Accepted in revised form 20 May 2000)

Summary

The characteristics of the respiration rates in precut green onion, as inuenced by oxygen levels and temperature, were examined to provide design factors for modied atmosphere packaging (MAP). Fresh-cut green onions (Allium stulosum L.) were prepared and sealed, with and without a CO2 absorbent, in gas-tight glass containers that had initially been purged with air or a gas mixture (O2 9%/N2 balance). The containers were stored at dierent temperatures (0, 10, 20 C). At 10 C, the maximum O2 uptake rate (Vm) and the O2 concentration at half-maximum (Km) uptake rate were 30.95 mL kg)1 h)1 and 1.63%, respectively. Regardless of temperature, the lower O2 limit was estimated to be about 1.0% O2 on the basis of respiratory quotient (RQ) increase. Respiration of cut green onion was dependent on O2 concentration as well as temperature, as shown by applying the MichaelisMenten type model and the Arrhenius equation. However, the presence of CO2 had little eect on O2 uptake of cut green onion at relatively high O2 concentrations (20%).
Lower oxygen limit, minimally processed vegetables, modied atmosphere packaging, respiration kinetics.

Keywords

Introduction

The demand for fresh-cut fruits and vegetables, both for retail and food service applications, has increased tremendously over the past few years. Such products provide both convenience and freshness to consumers, and attract the interests of many facets of the food industry including food manufacturers, retail food stores and restaurants. The preparation of these minimally processed products involves operations such as abrasion, peeling, cutting, shredding, as well as others (Wiley, 1994). Thus, the behaviour of the cut fruit and vegetable tissue is generally typical of that observed in plant tissues that have been wounded or exposed to stress conditions. These physiological changes include increased respiration, ethylene production, and susceptibility to oxidative brown-

*Correspondent: Fax: +82 342 709 9876; e-mail: sihong@kfri.re.kr; hsikfri@chollian.net

ing and entrance of microbes (King & Bolin, 1989). As a consequence, the fresh-cut products suer from rapid deterioration of quality and have a short shelf-life. The MAP can extend the shelf-life of the fresh produce including intact and cut commodities. Retardation of physiological deterioration in fruits and vegetables by low oxygen atmospheres with MAP is generally associated with depression of respiration rate. The relationship between the respiration rate and the O2 concentration in the surrounding atmosphere has therefore been a critical area of study in the physiology of many plant products. Successful characterization of these relationships would greatly facilitate the optimization of the MA packages (Cameron et al., 1989; Beaudry et al., 1992; Joles et al., 1994). Also of special interest is the lower limit for the O2 concentration within packages which marks the transition between aerobic and anaerobic respiration. The lower O2 limit has been determined by

2001 Blackwell Science Ltd

284

Respiration of fresh-cut green onion S.-I. Hong & D.-M. Kim

measuring the rate of CO2 production or contents of alcohol and other fermentative products at several O2 concentrations. However, it is timeconsuming and often dicult to evaluate the lower O2 limit owing to technical problems, including simultaneous determination of CO2 output and O2 uptake by crops (Gran & Beaudry, 1993). A variety of techniques have been used to investigate the respiration characteristics of fresh produce. A common method of measuring respiration involves a ow-through or open system where a gas stream is passed through a container of respiring fruits or vegetables at a known rate. By determining the dierences between the initial O2 and/or CO2 concentrations and those in the gas stream leaving the container, it is possible to calculate the rate of respiration (Lee et al., 1991). With this method, systematic errors may occur owing to diculties in measuring and controlling the ow rate. Also, because the changes occurring in the O2 concentration are small when compared to the large background concentration in air, it is generally easier to measure CO2 production. Permeable or MAP systems where plastic packaging bags are used to achieve steady state conditions have been used to measure respiration rate under dierent gas concentrations. As long as the area and permeability of the lm are known, the respiration rate can be calculated by measuring the partial pressure dierence between the package and the air. By varying the size of the packages and the weight of produce used, a range of atmospheres can be generated (Beaudry et al., 1992). When sample time can be arranged so that the packages reach a steady state, it can provide useful information. However, such data on respiratory behaviour is often required more urgently, and the method provides confusing results if the respiration rate of the commodity varies over the period of measurement (Bower et al., 1998). Static or closed systems can provide a convenient way of characterizing respiration of fresh produce using a single set of experiments. The changes in O2 and CO2 concentrations within a sealed container which result from respiration may be measured directly. While useful for short checks, such systems have some limitations, as changes occurring over extended periods of time are dicult to monitor continuously. For a CO2 intolerant produce, this method can not give

useful respiration data at the range of low O2 concentrations if it is necessary to maintain low CO2 concentration at the same time. Problems in static systems can be overcome by using two or more containers of initially dierent O2 concentrations with little CO2 (Yam et al., 1993; Lee et al., 1994). From two or multiple sets of gas concentration history data, the required information can be obtained over the O2 and CO2 ranges of interest, this is impossible by a typical static method. The purpose of this work was to test the hypothesis that investigating the respiration characteristics of precut green onion, as inuenced by oxygen concentration and temperature using a modied static system, would provide useful data that could be used for designing MA packages. Green onion was chosen for the investigation because of its importance in Asian vegetable production and consumption.
Materials and methods

Sample preparation Fresh green onions (Allium stulosum L.) were purchased from a local wholesaler in Seoul, Korea and held at 0 C until used. Green onions were sorted for obvious defects, trimmed, and washed with tap water. Trimmed stalks were crosscut into 10 cm length and the residual surface water was dried with paper towels. Characterization of respiration The respiration rates of precut green onion, as inuenced by O2 concentration and temperature, were measured by a modied closed system method (Yam et al., 1993; Lee et al., 1994). After equilibration at each temperature (0, 10, and 20 C) for 2 h, the cut pieces, weighing approximately 300 g, were placed into two 1.8 L glass jars with silicon sampling ports and were tightly sealed with metal clamps. The rst jar was closed immediately and the second jar was sealed after ushing with approximately 9% O2 at a ow rate of about 300 mL min)1 for 1 h. Gas samples were periodically analyzed using a gas chromatograph (Shimadzu GC 14-A, Tokyo, Japan) tted with a thermal conductivity detector. In some
2001 Blackwell Science Ltd

International Journal of Food Science and Technology 2001, 36, 283289

Respiration of fresh-cut green onion S.-I. Hong & D.-M. Kim

285

experiments, a sachet containing 10 g of Ca(OH)2 as a CO2 absorbent was put in the jars in an attempt to evaluate the CO2 eect on produce respiration. The respiration rates, O2 consumption and CO2 evolution, were calculated from linear regression curves of O2 decrease and CO2 increase at each sampling interval, and expressed in mL kg)1 h)1. All the results were presented as the averages of multiple experiments (N 3). Statistical signicance of mean values was examined by one-way analysis of variance at P < 0.05. The least squares method was used for linearregression analysis.
Results and discussion

Respiration of cut green onion as inuenced by O2 level Concentrations of O2 and CO2 in closed jars containing cut green onions at 20 C changed as shown in Fig. 1. In the rst jar which had been initially lled with air (Fig. 1a), there was a linear decline in O2 down to about 6% and a linear increase in CO2 up to 14%. After storage for 9 h, the rate of O2 decrease and CO2 increase changed, possibly as a result of the development of a partial vacuum and/or excessive CO2 accumulation in the jar. It is uncertain whether green onion is a CO2 tolerant product (Powrie & Skura, 1991). However, a CO2 content of more than 15% would probably alter the characteristic respiration of cut

green onion at low O2 concentration. In addition, numerous events of gas samplings as well as CO2 absorption could contribute to a partial vacuum in the jar. The second jar was initially ushed with a modied atmosphere of 9% O2 and N2 (Fig. 1b), and a similar pattern of O2 decrease and CO2 increase could be observed without CO2 accumulating to more than 10%. Interestingly, as the oxygen was consumed, O2 concentration in the jar with a CO2 absorbent showed a steeper decline than that without any CO2 absorbent, and then remained constant below 1%. The respiration rate is calculated as the dierence in O2 and CO2 concentrations between two consecutive measurements divided by the time. Average O2 concentration in this period is determined by obtaining the arithmetic mean. The respiration rates of cut green onion were aected by the O2 content within the jars as is shown in Fig. 2. In the rst jar at 20 C (Fig. 2a), the O2 consumption and CO2 evolution rates were maintained at 7080 mL kg)1 h)1 when average O2 concentrations were within the range of 920%. A signicant reduction (P < 0.05) in the respiration rates, however, was observed at O2 concentrations below 9%. At 56% O2, the O2 consumption rates from the jar with the CO2 absorbent nearly reached zero. For the jar without the CO2 absorbent, the respiratory quotient (RQ), determined by the ratio of CO2 production to O2 consumption, greatly increased at O2 concentrations below 5%. These probably can, at least

composition during storage at 20 C. (a) The rst jars initially lled with air, (b) the second jars initially ushed with the modied air of 9% O2/N2 balance; esh weight of green onions: 300 g, headspace volume: 1360 mL. Closed symbols: O2, open symbols: CO2, dd: without CO2 absorbent, ..: with CO2 absorbent. Vertical bars on each data point indicate standard deviation. International Journal of Food Science and Technology 2001, 36, 283289

Figure 1 Changes of gas concentration in the closed jars containing fresh-cut green onions with dierent initial gas

2001 Blackwell Science Ltd

286

Respiration of fresh-cut green onion S.-I. Hong & D.-M. Kim

Figure 2 Eect of O2 concentration on the respiration rate and the respiratory quotient of fresh-cut green onions with and

without CO2 absorbent in the closed jars at 20 C. (a) The rst jars initially lled with air, (b) the second jars initially ushed with the modied air of 9% O2/N2 balance. Closed symbols: O2 consumption rate, open symbols: CO2 production rate. ..: without CO2 absorbent; mm: with CO2 absorbent; : RQ. Vertical bars on each data point indicate standard deviation.

partly, be attributed to a high CO2 content and development of a partial vacuum in the rst jar, resulting in false respiration data. It is well known that a typical closed system using a single jar can not provide respiration rate data over a range of low O2 and CO2 concentrations (Lee et al., 1994). Nevertheless, no remarkable eect from CO2 (12%) on the respiration rates at high O2 concentrations (7%) was seen, the rates were nearly equal between the jars with and without the CO2 absorbent. The CO2 production, as well as O2 uptake rates, from the second jars with and without the CO2 absorbent decreased gradually as the O2 declined from 9 to 2%, and then reduced remarkably below 2% O2 (Fig. 2b). The O2 uptake rate for the jar with the CO2 absorbent nearly reached 0 at about 0.6% O2. At O2 concentrations below 1%, the RQ values for the jar without the CO2 absorbent began to increase and exceeded 1. The results are in agreement with previous reports (Leshuk & Saltveit, 1990; Kubo et al., 1996) related to respiration characteristics of fresh produce. It may indicate that a shift from aerobic to anaerobic respiration occurred at about 1% O2 and this concentration could be regarded as the lower O2 limit to aerobic respiration of fresh-cut green onion. When O2 concentration drops to less than the lower limit, there would be substantial accu-

mulation of ethanol and acetaldehyde, which are toxic to plant cells and cause o avours in fresh produce. For the jars stored at 0 and 10 C, the respiration of cut green onion showed the same pattern as that at 20 C, although the respiration rates were dierent depending on temperature (data not shown). Assuming that the respiration rate is a function of O2 concentration, applying a model based on MichaelisMenten type enzyme kinetics (equation 1) to the respiration of precut green onion (Lee et al., 1991) enabled us to obtain model parameters such as Vm, Km by simple linear regression analysis of equation 2. R Vm O2 Km O2 1

R respiration rate for samples (mL kg)1 h)1), Vm maximum respiration rate (mL kg)1 h)1), [O2] oxygen concentration (%, v/v), Km oxygen concentration at half Vm (%, v/v). Equation 1 can be transformed to the following linear form. 1 1 Km 1 R Vm Vm O2 2

As mentioned previously, the respiration data from single jars were inappropriate for a kinetic model. Thus, the respiration data from both the

International Journal of Food Science and Technology 2001, 36, 283289

2001 Blackwell Science Ltd

Respiration of fresh-cut green onion S.-I. Hong & D.-M. Kim

287

rst and the second jars were combined to obtain exact respiration rates over the O2 range of 020%. The results are plotted as 1/R against 1/[O2] (data not shown), giving Vm and Km. Because even minor errors on small values lead to large discrepancies when inversely transformed, only O2 concentrations above 0.65% were used in the regression analysis. The inversely transformed data were signicantly correlated linearly (P < 0.05). Respiration model parameters of fresh-cut green onion obtained from a modied closed system are presented in Table 1. Regardless of temperature, the values of Vm and Km for O2 consumption are consistently larger than those for CO2 production. Vm values increase with temperature while Km values have no dependence on temperature. This is in agreement with a previous report (Joles et al., 1994) which showed that the Km of raspberry fruit remained constant with changing temperature. The Vm and Km values for O2 consumption with and without the CO2 absorbent were similar. This indicates that the presence of CO2 (lower than 12  15% depending on temperature) had little eect on the O2 uptake of cut green onion at high O2 concentrations (20%). It is of interest that both Vm and Km vary greatly depending on species, cultivar, maturity, and storage conditions even within a commodity. For example, Lee & Lee (1996) reported that Vm and Km values of cut green onion (Allium wakeyi) for CO2 production at 10 C were 48.90 mL kg)1 h)1 and 0.46% O2 while those of other cut green onion (Allium stulosum) were 43.39 mL kg)1 h)1 and 1.24%. Lee et al. (1994) also found
Table 1 Respiration model

that prepared vegetables such as fresh-cut carrot, garlic, and cucumber at 10 C had Km values of 1.15%, 2.29%, and 4.38% for O2 consumption, respectively. Principally, the MichaelisMenten model is only valid for systems including one enzyme and one substrate. Therefore, the values of Vm and Km should be viewed with some caution, but they do provide a useful indication of how a mass of fresh produce responds to diminishing O2 concentration. Although a model like the MichaelisMenten is a very simple approach to a complex process such as respiration, it can be applied to various products without deep knowledge of the underlying biochemical and physiological reactions. The parameters obtained from the model can be used to predict respiration rates and simulate certain equilibrium O2 and CO2 concentrations inside a permeable package. Several authors have developed models to predict the gas composition in MAP based on respiration and permeability (Hayakawa et al., 1975; Cameron et al., 1989). Respiration of cut green onion as inuenced by temperature Temperature is the most important environmental factor in the postharvest life of fresh produce because of its dramatic eect on rates of biological reaction including respiration. Within the physiological temperature range (040 C), the respiration rate of fresh fruits and vegetables generally increases two- to three-fold for every 10 C rise in temperature (Van't Ho rule). As presented in Table 1, the Vm values of fresh-cut green onion
Vm (mL kg)1 h)1) 91.74 78.74 89.28 30.95 27.02 32.25 11.75 9.19 12.04 Km (% O2) 2.39 1.29 2.32 1.63 1.35 0.96 1.77 1.34 1.98 r2* 0.984 0.973 0.927 0.917 0.919 0.880 0.959 0.907 0.935

parameters of fresh-cut green onion obtained by a closed system method at various temperatures

Temp. (C) 20

Respiration type O2 consumption CO2 production O2 consumption (with CO2 absorbent) O2 consumption CO2 production O2 consumption (with CO2 absorbent) O2 consumption CO2 production O2 consumption (with CO2 absorbent)

10

*Linear regression analysis was carried out using the least square method.

2001 Blackwell Science Ltd

International Journal of Food Science and Technology 2001, 36, 283289

288

Respiration of fresh-cut green onion S.-I. Hong & D.-M. Kim

for O2 consumption as well as CO2 production also increased approximately three-fold with the increase in temperature from 0 to 20 C at 10 C intervals. In other words, the Q10 values for cut green onion are 2.73.0 in this temperature range. The Q10 concept allows calculation of expected respiration rates from a known rate. However, Q10 values for respiration rates of vegetables can vary over temperature ranges (Kader, 1987). The Arrhenius relationship can also consistently describe the dependence of the biological reactions, including respiration of fresh produce, on temperature. Plotting Loge Vm versus reciprocal of temperature showed a good linear t (r2 0.997 0.999, P < 0.05), following closely an Arrhenius behaviour (Fig. 3). The kinetic parameters for the respiration of fresh-cut green onion were derived

from the Arrhenius plot and given in Table 2. The frequency factor (k1) and activation energy (Ea) for respiration with and without CO2 absorbent were very similar. Haggar et al. (1992) reported that universal activation energy might be used for respiration in both air and modied atmospheres. The activation energy values of 66.5 71.3 kJ mol)1 for cut green onion are larger than those for other fresh vegetables including broccoli (62.766.1), cauliower (21.248.2), and green pepper (48.757.3), although the Vm values at corresponding temperature were much smaller than those of other vegetables (Yam & Lee, 1995). Therefore, cut green onion has a greater temperature dependence on respiration. No marked eect of temperature on the lower O2 limit was apparent, as shown in Fig. 4. If the lower O2 limit is considered to be the O2 concentration that causes a 50% increase in the RQ relative to the aerobic RQ (i.e., 1.5), then the lower O2 limit of fresh-cut green onion is estimated to be about 1.0% O2, regardless of temperature. Kubo et al. (1996) also found that some vegetables including cucumber, eggplant, sweet potato, broccoli, and cauliower had the lower O2 limit of 1%. In general, the lower O2 limits, determined by an RQ rise, have been observed to increase with increasing temperature for various fruits (Beaudry et al.,

Figure 3 Dependence of the maximum respiration rate (Vm)

of fresh-cut green onions on temperature. Closed symbols: O2 consumption, open symbols: CO2 production. ..: without CO2 absorbent, mm: with CO2 absorbent.

Table 2 Kinetic parameters for respiration of fresh-cut

green onion

Respiration type O2 consumption CO2 production O2 consumption (with CO2 absorbent)

1

k1 (mL kg)1 h)1) Ea (kJ mol)1) 1.31 1014 4.11 1014 6.45 1013 68.23 4.191 71.33 0.33 66.53 0.36

r2 0.997 0.999 0.999

Figure 4 Eect of O2 concentration on the respiratory

Standard error (P < 0.05).

quotient of fresh-cut green onions in the closed jars at various temperatures. d: 0 C; j: 10 C; r: 20 C.

International Journal of Food Science and Technology 2001, 36, 283289

2001 Blackwell Science Ltd

Respiration of fresh-cut green onion S.-I. Hong & D.-M. Kim

289

1992; Beaudry & Gran, 1993; Gran & Beaudry, 1993; Joles et al., 1994). For many fruits, resistance to O2 movement into the tissues is greatest at the skin (Cameron & Yang, 1982). Beaudry & Gran (1993) hypothesized that as the temperature increased, the rate of O2 consumption by the tissue rose more rapidly than skin permeability to O2. Thus, the O2 gradient across the skin increased as the temperature increased, leading to a rise in the lower O2 limit. However, it is uncertain whether this hypothesis is applicable to vegetables with dierent anatomical structures to fruits. In conclusion, this study reports the respiration characteristics of fresh-cut green onion as aected by O2 level and temperature. The respiration rate of cut green onion correlated well with the MichaelisMenten model and the Arrhenius equation, showing a signicant correlation with O2 concentration and temperature. In contrast, respiration showed no dependency on CO2 within a certain range. The respiration model parameters and kinetic data would be useful aids in designing an eective packaging system for prepared products such as cut green onion.
Acknowledgments

Research has been supported by a grant (HMP98-F-30008) of the Good Health R & D project, Ministry of Health & Welfare, R.O.K.
References
Beaudry, R.M., Cameron, A.C., Shirazi, A. & DostalLange, D.L. (1992). Modied atmosphere packaging of blueberry fruit: eect of temperature on package oxygen and carbon dioxide. Journal of American Society for Horticultural Science, 117, 436441. Beaudry, R.M. & Gran, C.D. (1993). Using a modied atmosphere packaging approach to answer some postharvest questions: factors inuencing the lower oxygen limit. Acta Horticulturae, 326, 203212. Bower, J.H., Jobling, J.J., Patterson, B.D. & Ryan, D.J. (1998). A method for measuring the respiration rate and respiratory quotient of detached plant tissues. Postharvest Biology and Technology, 13, 263270. Cameron, A.C., Boylan-Pett, W. & Lee, J. (1989). Design of modied atmosphere packaging systems: modeling oxygen concentrations within sealed packages of tomato fruits. Journal of Food Science, 54, 14131416. Cameron, A.C. & Yang, S.F. (1982). A simple method for the determination of resistance to gas diusion in plant organs. Plant Physiology, 70, 2123.

Gran, C.D. & Beaudry, R.M. (1993). Determination of the low oxygen limit for several commercial apple cultivars by respiratory quotient breakdown. Postharvest Biology and Technology, 3, 259267. Haggar, P.E., Lee, D.S. & Yam, K.L. (1992). Application of an enzyme kinetics based respiration model to closed system experiments for fresh produce. Journal of Food Process Engineering, 15, 143157. Hayakawa, K.I., Henig, Y.S. & Gilbert, S.G. (1975). Formulae for predicting gas exchange of fresh produce in polymeric lm package. Journal of Food Science, 40, 186191. Joles, D.W., Cameron, A.C., Shirazi, A. et al. (1994). Modied atmosphere packaging of `Heritage' red raspberry fruit: respiratory response to reduced oxygen, enhanced carbon dioxide, and temperature. Journal of American Society for Horticultural Science, 119, 540545. Kader, A.A. (1987). Respiration and gas exchange of vegetables. In: Postharvest Physiology of Vegetables (edited by J. Weichmann). Pp. 2544. New York: Marcel Dekker. King, A.D. Jr & Bolin, H.R. (1989). Physiological and microbiological storage stability of minimally processed fruits and vegetables. Food Technology, 43, 132135. Kubo, Y., Inaba, A. & Nakamura, R. (1996). Extinction point and critical oxygen concentration in various fruits and vegetables. Journal of Japan Society for Horticultural Science, 65, 397402. Lee, D.S., Haggar, P.E., Lee, J. & Yam, K.L. (1991). Model for fresh produce respiration in modied atmospheres based on principals of enzyme kinetics. Journal of Food Science, 56, 15801585. Lee, J.J. & Lee, D.S. (1996). A dynamic test for kinetic model of fresh produce respiration in modied atmosphere and its application to packaging of prepared vegetables. Food Science and Biotechnology, 5, 343348. Lee, D.S., Lee, K.S., Park, I.S. & Yam, K.L. (1994). Analysis of respiration characteristics of low CO2 tolerance produces for designing modied atmosphere package. Food Science and Biotechnology, 3, 99103. Leshuk, J.A. & Saltveit Jr M.E. (1990). A simple system for the rapid determination of the aerobic compensation point of plant tissue. Hortscience, 25, 480482. Powrie, W.D. & Skura, B.J. (1991). Modied atmosphere packaging of fruits and vegetables. In: Modied Atmosphere Packaging of Food (edited by B. Ooraikul & M.E. Stiles). Pp. 169245. New York: Ellis Horwood. Wiley, R.C. (1994). Introduction to minimally processed refrigerated fruits and vegetables. In: Minimally Processed Refrigerated Fruits and Vegetables (edited by R.C. Wiley). Pp. 114. New York: Chapman & Hall. Yam, K.L., Haggar, P.E. & Lee, D.S. (1993). Modeling respiration of low CO2 tolerance produce using a closed system experiment. Food Science and Biotechnology, 2, 2225. Yam, K.L. & Lee, D.S. (1995). Design of modied atmosphere packaging for fresh produce. In: Active Food Packaging (edited by M.L. Rooney). Pp. 5573. Glasgow: Chapman & Hall.

2001 Blackwell Science Ltd

International Journal of Food Science and Technology 2001, 36, 283289