Super-cooling phenomena in fruits, vegetables and seafoods

Christian James, Pierre Hanser, Stephen J James,

Food Refrigeration and Process Engineering Research Centre (FRPERC), The Grimsby Institute of Further & Higher
Education (GIFHE), HSI Building, Origin Way, Europarc, Grimsby, North East Lincolnshire, DN37 9TZ, UK
(jamesc@grimsby.ac.uk)

ABSTRACT
Super-cooling is the phenomena where the temperature of a solution or food material is reduced below its
freezing point without ice crystallization occurring, due to an energy barrier that must be surmounted before
nucleation starts. When ice crystallization begins the temperature is raised to the freezing point. The point at
which nucleation is initiated may be referred to as the ‘nucleation point’ or ‘metastable limit temperature’
(Cox & Moore, 1997).
Recent studies at FRPERC, as presented in this paper, have found significant (as low as -14.6°C for some
vegetables), and surprisingly stable, super-cooling to occur in a wide variety of vegetables (such as garlic,
shallots, peppers, broccoli and cauliflower). These studies have also demonstrated that some vegetables
(such as garlic, shallots and peppers) can be stored at temperatures significantly below their freezing point (as
low as -10°C for some products) for weeks without freezing occurring. Further studies, also presented in this
paper, have begun to investigate super-cooling in seafoods and have shown similar phenomena in products
such as prawns. This phenomena has implications in both chilled, super-chilled and frozen storage of such
products.

Keywords: Cold-chain; Super-cooling; Freezing; Freezing point

INTRODUCTION
This work was initiated following an opinion sought regarding the freezing of garlic bulbs (Allium sativum
L.). A company had reputably been importing ‘frozen’ garlic bulbs at a temperature of -6°C, however the
UK Customs questioned whether it was ‘frozen’. An independent report by another research organisation
had made an assumption that freezing would cause clear and obvious changes to the colour and structure of
the cells in the garlic cloves. They did not find this in garlic cloves that they inspected, hence they concluded
that the samples had not been previously frozen. However, a short experimental study carried out by the
authors indicated that the degree of freezing, and potentially damage, within a clove of garlic was not only a
function of temperature but is also influenced by the rate of temperature reduction and possibly by the length
of storage time. The study also found unusual super-cooling characteristics that indicated that garlic cloves
may not always be in a frozen state at a temperature of -6°C. This initiated further studies into the super-
cooling characteristics of garlic (James et al., 2009). Further research, reported in this paper, has shown that
many other vegetables show significant super-cooling and that shallots in particular have very similar super-
cooling characteristics to garlic.
References to super-cooling in solid foods are spread far and thin, most being in regard to fruits and
vegetables. Many of these published studies regarding super-cooling in fruits and vegetables have been
concerned with these phenomena in growing plants (Pearce, 2001) and its role in frost damage and frost
resistance. As early as the 1920s, Diehl (1924) reported that isolated apples could sometimes be cooled to a
point as low as 7 or 8 degrees below their freezing point (4°C of super-cooling) without ice formation,
provided the fruit was left undisturbed. Lucas (1954) cited other early studies showing 3°C and 1.5°C of
super-cooling in grapes and navel oranges, while his own study on lemons reported nucleation points of
between -3.9 and -4.4°C in whole fruits cooled in air, while a nucleation point of -6.1°C was reported in
whole lemons cooled in alcohol. Potatoes have been shown to super-cool for a short time to -3.9°C, about
3°C below their freezing point (Hruschka et al., 1961). Super-cooling of tomatoes was used as an example in
a more recent patent on a process for super-cooling by Cox & Moore (1997). Two individual, unreplicated,
trials showed that two fresh tomatoes had nucleation points of -4.3 and -4.5°C. In a separate experiment no
evidence of ice crystallization was apparent on visual inspection of the interior of a tomato cooled to an
internal temperature of -3.8°C in a ‘water’ bath at -4°C. Fuller & Wisniewski (1998) measured ten
independent nucleation events on the surface of a cauliflower curd cooled to -10°C at a rate of 15°Ch-1,
ranging from -6.5°C to -9.5°C. Super-cooling in frozen strawberries and its affect on ice crystal size and
formation has been investigated by Martins & Lopes (2007). Their data showed a difference between
nucleation and freezing point temperatures measured on the surface or centre of strawberries.
The aims of the experiments reported in this paper were to establish: (1) the freezing point of a variety of
vegetables, in particular garlic and shallots; (2) the degree and stability of super-cooling in a variety of
vegetables, in particular garlic and shallots.

MATERIALS & METHODS

1. Vegetable trials
Fresh broccoli, carrots, cauliflower, garlic, leeks, parsnips and shallots were obtained from a local
supermarket and stored in ambient conditions for no more than a week before use.
A series of experiments were initially conducted to establish freezing curves under slow freezing conditions.
Nucleation (metastable limit) temperatures during freezing were also recorded.
Single point metal shrouded hypodermic T-type (copper-constantan) probes (1mm diameter, 50mm length;
Electronic Temperature Instruments Ltd, UK) were used to measure temperatures in the geometric centre of
each of the six individual vegetable samples at a time. The six samples were placed in an expanded
polystyrene box (220 by 220 by 230mm, 20mm thick walls) and surrounded by small spheres (approximately
between 4 and 6mm diameter) of expanded polystyrene insulation (the aim being to slow the freezing and
thawing rates to allow easier interpretation of the temperature plots). The box was placed in a standard
domestic chest freezer (operating at -30±1°C). Temperatures were recorded every 30 s using a datalogger
(Eltek Type 1000, Grant Instruments (Cambridge) Ltd, UK). When all the probe temperatures in the samples
were reading below -10°C, the freezing was stopped. A total of ten replicate experiments were carried out.
The freezing point was interpreted from the freezing curve, as carried out in previous studies by the authors
(James et al., 2005; James et al., 2009).
Further trials were conducted to establish the nucleation (or metastable) limit temperature of the garlic cloves
and shallot bulbs by storing the bulbs at temperatures within the super-cooling temperature range identified
in the initial freezing trials.
Trials were carried out in an experimental temperature controlled wind tunnel (James et al., 1979) under
static air conditions (<0.5 ms-1). The initial experimental run was carried out -5±0.5°C, in each subsequent
run the air temperature was progressively decreased by 1°C increments per experimental run until all samples
were found to freeze, i.e. the temperature condition exceeded the nucleation (metastable limit) temperature.
New samples were used in each trial. Three samples of each vegetable being used for each experimental run.
To further confirm the effect of long term storage in a super-cooled state on the visual appearance of garlic
and shallots, whole shallot and garlic bulbs and individual unpeeled garlic cloves were stored, un-covered on
metal trays, for one week under four different environmental conditions: ambient, chilled (1±1°C), super-
cooled (-6±0.5°C) and frozen (-30±0.5°C), at static air velocities. Temperature data were recorded for the
super-cooled and frozen samples. After one week of storage samples were compared visually.

2. Preliminary seafood trials

Preliminary trials have begun to investigate super-cooling in seafoods. Initial trials have looked at samples
of previously frozen peeled Atlantic prawns (Pandalus borealis). A similar method to that described
previously is being used, but using a domestic freezer (Whirlpool AFG522-C/H) modified by the addition of
internal fans to improve air circulation and thus temperature uniformity, and internal heaters controlled by a
Eurotherm 452 controller using on/off control with a 0.1°C deadband to act against the freezer refrigeration
to maintain desired storage temperatures within ±0.5°C of the set-point.

RESULTS & DISCUSSION

Vegetable trials
Significant super-cooling was seen in most of the freezing curves of all of the vegetables prior to nucleation
(Table 1). Super-cooling was observed in all of the 15 garlic and all of the 10 shallot samples, 9 of the 10
broccili, carrot and leek samples and 7 of the 10 cauliflower samples. However, super-cooling only occurred
in 4 of the 10 parsnip samples. The greatest overall degree of super-cooling was apparent in garlic samples,
where super-cooling was on average 10.3 degrees lower than the initial freezing point (-2.7°C). The second
greatest overall degree of super-cooling was apparent in cauliflower samples, where super-cooling was on
average 3.8 degrees lower than the initial freezing point (-1.5°C). Although in one case a nucleation point
temperature of -8.3°C was recorded, in general nucleation point temperatures were below -5.2°C. However,
super-cooling was only detected in 7 of the 10 cauliflower samples frozen, and there was a great degree of
variation in super-cooling temperatures. The minimum recorded nucleation point temperature was -1.9°C.
In shallot samples super-cooling was observed in all samples, with the minimum degree of super-cooling
being 1.1 centigrade degrees below the freezing point. Mean nucleation temperatures were 3.3 degrees
below the initial freezing point (-1.6°C).
Table 1. Freezing and nucleation points (to 1 d.p.) of vegetable samples, frozen at -30±1°C (n=10, *n=15)
Freezing Nucleation temperature * Degrees of under-cooling *
Occurrences
point (°C) (°C)
Vegetable of super-
Mean Mean
cooling
(SD) Min Max (SD) Min Max Mean
Broccoli -2.1 (0.3) 9/10 -2.3 -9.8 -4.4 (2.4) 0.4 7.7 2.5 (2.3)
Carrot -1.6 (0.6) 9/10 -1.3 -3.9 -2.7 (0.8) 0.4 2.6 1.1 (0.7)
Cauliflower -1.5 (0.3) 7/10 -1.9 -8.3 -5.2 (2.6) 0.6 6.9 3.8 (2.6)
Garlic* -2.7 (0.3) 15/15 -7.7 -14.6 -13.0 (1.7) 4.9 12.0 10.3 (1.6)
Leek -1.9 (0.3) 9/10 -2.3 -3.9 -3.3 (0.6) 0.6 2.3 1.6 (0.6)
Parsnip -2.2 (0.2) 4/10 -2.6 -3.1 -2.8 (0.2) 0.4 0.9 0.7 (0.2)
Shallot -1.6 (0.2) 10/10 -2.7 -7.3 -5.4 (1.4) 1.1 4.7 3.3 (1.3)
*when super-cooling occurred

Figure 1. Comparison of unpeeled garlic cloves after storage for one week at (from left to right) ambient, chilled
(1±1°C), supercooled (-6±0.5°C) and frozen (-30±0.5°C) temperatures, respectively.

Figure 2. Comparison of unpeeled shallot bulbs after storage for one week at (from left to right) ambient, chilled
(1±1°C), super-cooled (-6±0.5°C) and frozen (-30±0.5°C) temperatures, respectively.

Peeled garlic cloves held in the experimental temperature controlled wind tunnel under static air conditions
(<0.5 ms-1) at temperatures between -6 and -9°C±0.5°C did not freeze, despite regular spikes in the
environmental temperature caused by defrosts. The cooling curves showed no signs of nucleation or a
freezing plateau. When examined visually after between 16.5 to 69 h of storage at these temperatures there
was no external or internal sign of freezing damage in any of the peeled garlic cloves, and cloves were
considered visually identical to non-frozen cloves. Freezing did occur in peeled garlic cloves held in the
experimental temperature controlled wind tunnel under static air conditions (<0.5ms-1) at temperatures
between -10°C and -13°C±0.5°C, however its occurrence was variable. Out of 5 cloves stored at -10±0.5°C
for approximately 24 h only one froze (after 185 min). In two runs at -13±0.5°C, 4 out of 5 cloves and 2 out
of 5 cloves did not freeze after 46.5 and 45 h storage, respectively.
Whole unpeeled shallot bulbs held in the experimental temperature controlled wind tunnel under static air
conditions (<0.5 ms-1) at temperatures between -5 and -6°C±0.5°C did not freeze. The cooling curves
showed no signs of nucleation or a freezing plateau. When examined visually after between 24 h storage at
these temperatures there was no external or internal sign of freezing damage in any of the unpeeled shallot
bulbs, and bulbs were considered visually identical to non-frozen bulbs. Freezing did occur in unpeeled
shallot bulbs held in the experimental temperature controlled wind tunnel under static air conditions (<0.5ms -
1
) at temperatures at -7°C, however its occurrence was variable. Out of 5 bulbs stored at -7±0.5°C for
approximately 24 h only 2 froze (after 120 min and 527 min). However, all 5 samples held at -8°C froze.
To further confirm the effect of long term storage in a super-cooled state on the visual appearance of garlic
and shallot bulbs and cloves, unpeeled garlic cloves, whole garlic bulbs, and unpeeled whole shallot bulbs
were stored, un-covered on metal trays, for one week under four different environmental conditions: ambient,
chilled (1±1°C), super-cooled (-6±0.5°C) and frozen (-30±0.5°C), at static air velocities. Temperature data
were recorded for the super-cooled and frozen samples. After one week of storage samples were compared
visually. There was no difference in external or internal appearance of unpeeled garlic cloves (Figure 1),
whole garlic bulbs, and unpeeled whole shallot bulbs (Figure 2) that had been stored under ambient, chilled
(1±1°C) or super-cooled (-6±0.5°C) conditions, however there were clear signs of unacceptable visual
damage to bulbs that had been stored at -30±0.5°C.
There are two forms of ice nucleation, spontaneous (homogeneous nucleation) and that catalysed by another
substance (heterogeneous nucleation) (Pearce, 2001; Zachariassen et al., 2000). The lower limit for
homogeneous nucleation in plant materials is -38.5°C (Pearce, 2001). However homogeneous nucleation is
rare in biological systems since there are many intrinsic and extrinsic substances present in nature that can act
as heterogeneous nucleators (Pearce, 2001), including: (a) ice nucleation-active bacteria; (b) other biological
molecules and structures; and (c) organic and inorganic debris. Thus the factors that affect the degree of
super-cooling are not the same as those that determine the freezing point. Lucas’ (1954) states that the
amount of super-cooling is independent of rate of cooling, time at a given temperature, cell size, or osmotic
pressure. However other authors state that cell size and osmotic pressure are factors favouring super-cooling
(Fanyi et al., 2002). Other factors that favour super-cooling are low moisture content, little or no
intercellular space for nucleation, absence of internal nucleators, barriers against external nucleators, and the
presence of anti-nucleators (Pearce, 2001; Levitt, 1980; Fanyi et al., 2002). The role of extrinsic and intrinsic
nucleators in plants and nature has been reviewed in detail by Pearce (2001) and Zachariassen and
Kristiansen (2000).
One of the main extrinsic nucleators cited is surface moisture. Many authors report that ice nucleation can be
facilitated in tests of the frost hardiness of plants by spraying water on the surface of the plants prior to
freezing (Fuller and Wisniewski, 1998). Although Lucas (1954) did not show that the humidity of the
surrounding air or moisture on the fruit surface had any effect on the ice nucleation point in trials on detached
lemons. In the experiments described in this paper the peeled and unpeeled garlic cloves were stored in dry
ambient conditions before use so there was little, if any, surface moisture available to initiate nucleation thus
this may be a factor in the low super-cooling temperatures observed (although some condensation may have
occurred on introducing the warm garlic into the sub-zero environments used).
In many plants the most commonly reported intrinsic ice nucleators appear to be of bacterial origin (Fuller et
al., 1994). However, plants themselves also contain non-bacterial nucleating agents, some of which function
at relatively high temperatures (Pearce, 2001). The ability to supercool appears also to be genetic and
heritable (Fuller et al., 1994). It is thus possible that current work into biotechnological manipulation of
plants to improve frost hardiness may also result in the production of fruits and vegetables with increased
super-cooling characteristics. Environmental conditions during growing will also alter super-cooling
characteristics (Pearce, 2001). Being plant bulbs it may be assumed that garlic bulbs and cloves have some
inherited resistance to frost damage and that this translates in their ability to supercool, however we have
been unable to identify any studies into super-cooling in plant bulbs that substantiate this hypothesis.
The aims of the experiments reported in this paper were to establish: (1) the freezing point of a variety of
vegetables, in particular garlic and shallots; (2) the degree and stability of super-cooling in a variety of
vegetables, in particular garlic and shallots. Overall these results clearly indicate that garlic cloves and bulbs
and shallots can be stored at a temperature significantly lower that their freezing point without ice crystal
nucleation and formation. This has potentially significant implications to the long term storage of garlic,
shallots, and perhaps other vegetables. Refrigeration is important in both maintaining the safety and quality
of many foods and enabling food to be supplied to an increasingly urbanised world. It has been estimated
that 40% of all food requires refrigeration (Mattarolo, 1990, however, less than 10% of such perishable
foodstuffs are in fact currently refrigerated (Coulomb, 2008). It is estimated that post-harvest losses
currently account for 30% of total production (Coulomb, 2008). The production of food involves a
significant carbon investment that is squandered if the food is then not utilised. The International Institute of
Refrigeration (2009) estimate that, in theory, if developing countries could acquire the same level of
refrigerated equipment as that in industrialized countries, over 200 million tonnes of perishable foods would
be preserved, this being roughly 14% of the current consumption in these countries. The storage life of
vegetables at ambient temperature is significantly limited by microbial growth (causing spoilage), at chilled
temperatures (between 10°C to -2°C) microbial growth is limited but product will eventually spoil. At frozen
temperatures (<-2°C) microbial growth is prevented, but damage can occur to vegetable tissues through ice
formation. If super-cooled distribution could be used instead of frozen distribution for some fruits and
vegetables it could substantially reduce energy consumption whilst reducing food waste.

Preliminary seafood trials

Initial trials have shown significant super-cooling in previously frozen samples of cooked peeled Atlantic
prawns cooled slowly to temperatures as low -10°C (Table 2). Initial PCA analysis has yet to show any
relationship between physical parameters such as prawn size, water content and the degree of super-cooling,
however rate of cooling does appear to have some influence on the degree of super-cooling.

Table 2. Freezing and nucleation points (to 1 d.p.) of “jumbo” (mean 7.9 g) and “large” (mean 2.2 g) whole previously
frozen cooked peeled Atlantic prawns, frozen at -10±1°C (n=24)
Freezing Nucleation temperature * Degrees of under-cooling *
Occurrences
point (°C) (°C)
Size of super-
Mean Mean
cooling
(SD) Min Max (SD) Min Max Mean
“jumbo” -2.1 (0.1) 18/24 -7.9 -2.2 -5.9 (1.4) -6.0 -0.1 -3.6 (1.6)
“large” -1.9 (0.2) 15/24 -7.5 -4.0 -6.5 (1.0) -5.7 -2.0 -4.6 (1.2)

Preliminary trials on samples of cod (freezing point -1.4°C), herring (freezing point -3.6°C) and squid
(freezing point -2°C) have also shown that they are capable of super-cooling to -5.3, -9.2, and -8.6°C,
respectively. Further studies are on-going to establish the stability of super-cooling in such foods and the
factors that may govern super-cooling in these particular products.

CONCLUSION
Our studies have found significant, and surprisingly stable, super-cooling to occur in a wide variety of
vegetables (particularly garlic and shallots) and have demonstrated that some vegetables (such as garlic and
shallots) can be stored at temperatures significantly below their freezing point for weeks without freezing
occurring. This has potentially significant implications to the long term storage of such vegetables, and
perhaps other vegetables and fruits. It also shows from a legal perspective that some products stored and
distributed at temperatures below their freezing point may not be frozen. Being plant bulbs it may be
possible that garlic bulbs and cloves have some inherited resistance to frost damage and that this translates in
their ability to supercool, however we have been unable to identify any studies into super-cooling in plant
bulbs that substantiate this hypothesis. Preliminary studies are now also showing that significant, though at
present less stable, super-cooling also occurs in seafoods, particularly prawns, subjected to similar cooling
conditions to those investigated previously for vegetables. Since the composition of such products is
different it is more likely that super-cooling is related to external factors, such as cooling rate and external
temperature, than to intrinsic factors.

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