CHEMISTRY/BIOCHEMISTRY

Chemical Characteristics and

Storage Stability of Pickled Garlic
Prepared Using Different Processes
L. REJANO, A.H. SANCHEZ, A. de CASTRO and A. MONTANO

ABSTRACT A second lot of peeled garlic cloves was

Three processes for preparing peeled pickled garlic were studied: (P) packing blanched in water at 90°C for 15 min and
directly with an acidified brine, followed by pasteurization (90°C, 8 min); (B) blanch- then packed. A portion of blanched-garlic
ing with hot water (90°C, 15 min) before packing, followed by a preservation treat- cloves was packed into 10 “8 Par” glass bot-
ment (pasteurization, preservatives + refrigeration); and (MB) same as B, except tles with the acidified cover brine used pre-
that microwave blanching was used (120g garlic cloves, 825W, 1.5 min). All prod- viously (added at z70°C). Five bottles were
ucts were microbiologically stable but fructans were hydrolyzed during storage pasteurized at 90°C for 5.5 min and the rest
at 27°C. A green pigment was also apparent in (P). Products from (B) or (MB) had left as non-pasteurized control. Both prod-
better quality than that from (P). Most stable and best quality products resulted ucts were stored at 27 ± 2°C. The remaining
from the blanching treatments and water blanching resulted in the product with portion of blanched garlic cloves was also
best color. packed with the same cover brine, except it
Key Words: garlic, pickles, blanching, storage, preservation was fortified with 0.27% potassium sorbate
and 0.24% sodium benzoate, and maintained
under refrigeration (4°C) during storage. The
INTRODUCTION addition of the starter, and to inactivate soft- third lot of peeled garlic cloves was blanched
PICKLED GARLIC HAS BEEN MARKETED ening enzymes (Fleming et al., 1983). using a domestic microwave oven (maximum
for direct consumption, similar to other veg- Blanching with boiling water for 15 min has power output 1,100W, 2450 MHz, Moulin-
etables such as cucumbers, olives or capers. been used for deactivating alliinase before ex, France). Separate 120g garlic clove batch-
However, in contrast to those products, pub- determination of alliin in garlic products es were heated on a microwavable plate at
lications related to the preparation or use of (Mochizuki et al., 1988). Blanching with 75% power for 90s. The garlic cloves were
pickled garlic are very few. Kim et al. (1994) microwave energy has been reported to have then packed and stored in the same way as
prepared pickled garlic, submerging the advantages over conventional hot water or the water blanched product.
peeled cloves in 2% acetic acid containing steam blanching by reduction of process
2% NaCl, with the aim of studying the cor- times, energy and water usage and improve- Sampling
relation between pungency and allicin con- ment of product quality (Ponne et al., 1994). Samples were taken before processing
tent during aging. Our objectives were to prepare pickled (raw garlic), immediately after blanching,
Pickled garlic is marketed to the consumer garlic using different processes (direct pack- and after 9d and 4 mo storage, for chemical
peeled and packed in glass containers in a ing, packing after blanching with hot water, analyses and determinations of firmness and
cover liquor (brine or vegetable oil) with or and packing after blanching with microwave color. In addition, microbiological analyses
without addition of other aromatic plants. energy) and to study changes in chemical were carried out at 4 mo storage. For chemi-
Usually, the flavor is notably free from the characteristics, firmness and color during cal analyses, raw or blanched garlic cloves
pungent flavor typical of raw garlic. The main preparation and storage. (100g) were mixed with an equal weight of
pungent component of raw garlic is attribut- water and homogenized using a mixer. After
ed to alk(en)yl thiosulfinates (mainly allicin) MATERIALS & METHODS 30 min incubation at room temperature
generated by the action of alliinase, which is (<20°C), the solid part was separated by cen-
activated on cutting or splitting the garlic Preparation of pickled garlic trifugation at 16,000 x g for 20 min using a
clove, on alk(en)yl cysteine sulfoxides (main- Fresh garlic bulbs were obtained from a Sorvall RC-5 superspeed refrigerated centri-
ly allyl cysteine sulfoxide or alliin) (Stoll and market in Seville (Spain). After cracking the fuge (Du Pont Instruments, Newton, CT). A
Seebeck, 1951; Block 1992). Consequently bulbs to separate the cloves and peeling the portion of supernatant was used for analysis
a treatment to deactivate alliinase seems to outer covering (Fig. 1), defective and small- of pH, titratable acidity, and NaCl. The rest
be fundamental for the preparation of this est cloves were discarded. The remaining was frozen (-30°C) until analysis by HPLC
type of product. peeled cloves were washed with tap water and GC. In the packed product, garlic cloves
Blanching with hot water, steam or elec- and separated into three lots. One lot was were mixed with an equal weight of cover
tromagnetic energy is widely used in the veg- directly packed into 10 “ 8 Par” glass bottles brine and homogenized and the homogenate
etable products industry. It helps eliminate (115g garlic cloves and 115 mL cover liquor was taken through the above procedure.
the enzymatic activity responsible for flavor capacity) using, as cover liquor, a brine (5%
alterations and tissue softening (Juliot et al., NACl) acidified with a lactic acid-acetic acid Chemical and microbiological
1989; Ponne et al., 1994). In vegetables sub- mixture. The concentration of each acid analyses
jected to controlled fermentation, a prior (0.74%, w/v) in the cover brine was calcu- Titratable acidity (by titration up to pH
blanching with hot water has been used to lated to give a pH of 4.0 after equilibrium 8.3 with 0.5N NaOH and expressed as % (w/
eliminate undesired microorganisms before between garlic cloves and brine. Half of the v) of lactic acid) and pH were determined
packed product (5 bottles) was pasteurized using a Metrohm 670 Titroprocessor (Heri-
for 8 min in a water bath at 90°C (cover brine sau, Switzerland). Sodium chloride was de-
was added at <70°C) and the remainder was termined by titration with AgNO3 (Femin-
The authors are affiliated with Instituto de la Grasa
(C.S.I.C.), Apartado 1078, 41012 Seville, Spain. left as non-pasteurized control. All bottles dez-Diez et al., 1985).
were stored in a room maintained at 27 ± 2°C. Water soluble carbohydrates (fructans,

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 Ethanol was analyzed by GC using the
headspace method described by Montano et
al. (1990). Its presence was confirmed by the
HPLC method for organic acids. Carbon di-
oxide was qualitatively analyzed by head-
space GC using a Fisons HRGC Mega 2 Se-
ries gas chromatograph (Fisons Instruments,
Milan, Italy) fitted with a hot wire detector
and a split-splitless injection port with heli-
um as carrier gas. The column was a GS-Q
fused-silica capillary column (30m x 0.53
mm i.d., J&W Scientific). The oven temper-
ature was 50°C and the detector temperature
was 100°C.
Allicin was determined by HPLC follow-
ing the method described by Kim et al. (1994)
with slight modifications. The system con-
sisted of a Waters 600E liquid chromatograph
fitted with a Rheodyne 7125 injector, a Wa-
Fig. 1—Experimental design of processing of pickled garlic
ters Model 994 photodiode array detector and
a Waters Model 5200 printer-plotter. A C18
mBondapak column (10 mm, 39 × 300 mm,
Waters Assoc.) was used for separation. Op-
erating conditions were: mobile phase, meth-
sucrose, glucose, fructose) were analyzed by 0.005M H2S04 as mobile phase at 0.7 mL/ anol (50%) in water; flow rate, 1.0 mL/min;
HPLC. The system consisted of a Hewlett- min. Detection was carried out using a Wa- room temperature; detection, 254 nm. To 0.5
Packard Series 1050 liquid chromatograph ters Model 994 photodiode array detector set mL of sample extract, diluted with water if
(Avondale, PA) fitted with a Rheodyne 7125 at 220 nm in series with a Perkin Elmer Mod- necessary, 0.5 mL of methanol was added.
injector (Cotati, CA), a Perkin Elmer Model el LC-25 RI detector. The first detector was The mixture was centrifuged at 11,600 x g
LC-25 refractive index (RI) detector (Nor- used for detection of citric acid without in- for 5 min and an aliquot (20 mL) injected
walk, CT) and a Hewlett-Packard Model terference from carbohydrates. The RI detec- into the chromatograph. Pure allicin, for an
3396 Series II integrator. An Aminex HPX- tor was used for detection of lactic and ace- external standard, was isolated from garlic
87C carbohydrate analysis column (Bio Rad tic acids. Samples (I mL) were acidified with juice following the procedure described by
Labs, Richmond, CA) held at 65°C was used. concentrated H2SO4 (10 mL) and, after cen- Yu and Wu (1989), except that a Spherisorb
Deionized water was used as eluent at 0.7 trifugation at 11,600 × g for 10 min, an ali- ODS-2 semipreparative column (250 × 10
mL/min. To 1 mL of sample extract was add- quot (20 mL) was injected into the chromato- mm, 5 mm, Supelco) was used.
ed 1 mL of deionized water plus 2g of a graph. Concentrations were calculated by Procedures for enumeration of Enterobac-
mixed anion/cation resin (Serdolit MB-3, comparison of peak heights with those of teriaceae, lactic acid bacteria and yeasts were
Serva Feinbiochemica GmbH & Co., Heidel- external standards for each compound. The as described by De Castro et al. (1995).
berg, Germany) which had been washed with identity of citric acid was confirmed by dem-
deionized water and then dried under vacu- onstrating its degradation by citrate lyase Quality evaluation
um to remove excess water. The mixture was from Enterobacter aerogenes (Sigma Chem- Firmness of garlic cloves was measured
shaken occasionally during a 60 min de- ical Co., St. Louis, MO). by means of an Instron test device (Model
salting period. An aliquot (<1 mL) of the so-
lution was centrifuged at 11,600 x g for 10
min and 20 mL were injected into the chro-
matograph. Fructans were eluted as a large Table 1-Changes in chemical characteristics during preparation and storage of fresh-pack
garlica
single peak at the solvent front. The presence
of interfering coeluting samples was insig- Fresh-pack Nonpasteurized
garlicb controlb
nificant. Peak area of fructans was compared
with that of an aqueous standard of inulin Chemical Raw 9 4 9 4
characteristicsc garlicd days months days months
from chicory root (Sigma) treated in the same
way. Quantification of simple sugars was also NaCl — 2.9 2.8 3.0 3.0
pH 6.33 4.01 3.95 3.95 4.04
based on peak area and performed by the ex- Acidity
ternal standard method using a standard mix- (% as lactic acid) 0.40 1.46 1.56 1.53 1.58
ture of sucrose, glucose, and fructose treated Fructans
in the same way as the sample. As an alter- (% as inulin) 10.3 19.6 11.4 12.7 8.9
Sucrose (%) 0.99 1.00 1.08 1.17 0.94
native HPLC procedure, also used to con- Fructose (%) 0.27 0.28 3.32 1.14 3.79
firm identity of single sugars, separation was Glucose (%) 0.13 0.09 0.29 0.13 0.38
done on a Spherisorb NH2, column (5 mm, Citric acid (%) 0.40 0.38 0.44 0.37 0.39
Lactic acid (%) ND e 0.40 0.47 0.45 0.51
25 cm x 0.4 cm i.d., Teknokroma, Barcelo- Acetic acid (%) ND 0.38 0.42 0.39 0.42
na, Spain); acetonitrile-water (75:25, v/v) Ethanol (%) ND ND ND 0.16 0.13
was used as eluent at 1 mL/min. Allicin (%) 0.17 Tr f ND 0.04 ND
aInitial cover brine contained NaCl (5%) and lactic and acetic acids (0.74% each)
Organic acids (citric, lactic, and acetic) b In a garlic/cover brine mixture (50/50, w/w)
were analyzed by HPLC using a cation-ex- c % = g compound/100 mL extract
d In a garlic/water mixture (50/50, w/w)
change column (Aminex HPX-87H, 300 x e ND = Not detected
7.8 mm i.d.) (BioRad Labs.) at 65°C, with f Tr = Trace amounts < 0.05%

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 Preparation of Pickled Garlic . . .

1011) using a Kramer shear compression cell. pasteurization (fresh-pack garlic), at 9 days (data not shown) revealed no microbial
The cross head speed was 200 mm/min. and 4 mo storage (Table 1). Fructans, poly- growth. In the non-pasteurized control this
Firmness was expressed as the mean of 10 mers with varying numbers of fructose mol- may be attributed to the presence of inhibi-
replicate measurements, each of which was ecules and a terminal sucrose, are the main tors in the garlic (Patel et al., 1985) or to the
performed on one garlic clove (total weight, water soluble carbohydrates of garlic, yield- fact that the initial epiphytic microflora, con-
4.0 ± 0.5g). Shear compression force was ing polymers with a degree of polymeriza- sisting mainly of Enterobacteriaceae (data
expressed as N/g of product. tion ranging up to 50 (Darbyshire and Hen- not shown), was destroyed by the pH (z4.0)
Colorimetric measurements were per- ry, 1981). For quantification of such com- inside the pack. Acidification of the cover
formed using a Color-View Model 9000 pounds we assumed that they presented the brine in brine-stock cucumbers resulted in
spectrophotometer (BYK-Gardner, Inc.) with same RI response as a standard of inulin from inhibition of Enterobacteriaceae (McDonald
a measurement area of 11 mm dia, 45° cir- chicory. Livingston (1990) found no signifi- et al., 1991).
cumferential illumination and an observation cant differences in RI response, based on In fresh-pack garlic the main change dur-
angle of 0°. The apparatus was previously peak area, between fructans isolated from ing storage was the hydrolysis of fructans,
calibrated with a white standard (L* = 98.80, oats or barley and inulin from chicory. Su- shown by the considerable increase in con-
a* = –0.01, b* = –0.08). All measurements crose was the main simple sugar, with a con- centration of fructose and glucose and the
were made on the CIE 1976 L*a*b* scale centration of 1% (w/v) in the aqueous ex- simultaneous decrease in fructans (Table 1),
using illuminating conditions CIE type C, tract of raw garlic (Table 1). Citric acid was calculated as 44%. The sucrose level, how-
10° observer. Results were expressed as the the main organic acid, with a concentration ever, remained almost constant. Hydrolysis
mean of ten replicate measurements, each of (0.4%) similar to that found in the endocarp of fructans has also been reported during the
which was performed on one garlic clove. of ripe cucumbers (McFeeters et al., 1982). storage of onions (Suzuki and Cutcliffe,
The allicin content in the raw garlic (0.17%) 1989). The non-pasteurized control showed
Statistical analysis was within the concentration range reported hydrolysis of fructans, with a decrease in
Data from packed blanched products were by lberl et al. (1990) in raw garlic from dif- concentration of z30%. In addition, an ap-
examined by analysis of variance using STA- ferent sources. Moisture content of raw gar- preciable amount of ethanol was detected, to-
TISTICA software (StatSoft Inc., 1996). lic was z67%, deduced from the concentra- gether with the formation of abundant gas
Wherever F values were significant, Dun- tion (2.8-3.0%) of NaCl measured after pack- (CO2) on opening the bottles. Apparently, the
can’s Multiple Range Test was used to sepa- ing. In both the pasteurized (fresh-pack gar- lack of oxygen inside the pack induced anaer-
rate of mean effects. Significance was defined lic) and control product, the pH and titrat- obic respiration or fermentative metabolism
at p < 0.05. able acidity were z4.0 and 1.5% (as lactic (Taiz and Zeiger, 1991). The amount of alli-
acid) respectively. The slight increase in ti- cin in the packed product, both pasteurized
RESULTS & DISCUSSION tratable acidity observed during storage has and control, was considerably less than that
been quite common in pickled vegetable of raw garlic, and was undetected after 4 mo
Fresh-pack garlic products (Fleming et al., 1983) and is not storage. Kim et al. (1994) found a gradual
Chemical characteristics of raw garlic related to microbial growth. Microbiologi- decrease in the concentration of allicin in
were compared with those after packing and cal analyses carried out after 4 mo storage pickled garlic with storage time (at 20°C),

Table 2—Czhanges in color and firmness during preparation and Table 4—Main effects of blanching, preservation treatment and stor-
storage of fresh-pack garlic age time on color and firmness of packed blanched garlica
Fresh-pack Nonpasteurized Color parameter
garlicb controlb
Main effects L* a* b* Firmness (N/g)
Raw 9 4 9 4
Parameter garlicd days months days months Blanching
Hot water 68.6a -3.3a 13.2a [Link]
L* 80.6 68.1 65.4 44.5 58.2 Microwave 67.9a -2.2b 11.8b 26.8b
a* -1.9 -7.3 -2.7 -16.9 -1.7 Preservation treatment
b* 21.8 15.6 26.1 11.0 30.3 Control 66.7a - 2.4a [Link] 24.8a
Firmness (N/g) 72.0 46.5 45.7 73.9 74.4 Pasteurization 68.8b -2.4a 13.2a 22.4a
Preservatives 69.3b -3.3b 1 1. 1 b 24.5a
Storage time
9 days 69.1 a -3.2a 10.6a 25.4a
4 months 67.4b -2.2b 14.4b 22.4b
a Means with different letters within a column for each effect are different (p < 0.05)

Table 3—Main effects of blanching, preservation treatment and storage time on chemical characteristicsb of packed blanched garlica
Main effects pH Acidity (%) Fructans (%) Sucrose (%) Fructose (%) Glucose (%) Citric acid (%)
Blanching
Hot water 3.97a 1.45a 9.9a 0.81a 1.12a 0.13a 0.34a
Microwave 4.05a 1.55b 11.6b 0.96b 0.98a 0.14a 0.39b
Preservation treatment
Control 3.97a 1.50a 10.4a 0.91a 1.54a 0.17a 0.37a
Pasteurization 3.99a 1.49a 9.9a 0.91a 1.43a 0.17a 0.38a
Preservatives 4.06a 1.51a 12.0a 0.83b 0.18b 0.07b 0.34b
Storage time
9 days 4.00a 1.46a 12.2a 0.87a 0.24a 0.08a 0.35a
4 months 4.02a 1.54a 9.3b 0.90a 1.86b 0.19b 0.38b
a Means with different letters within a column for each effect are different lp - <0.05).
b Means values for NaCi, lactic acid, and acetic acid were 2.8%, 0.41%, and 0.40%, respectively; effects of blanching, preservation treatment and storage time on these characteristics
were not significant (p<0.05).

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 until its total disappearance after 60d. the parameters L*, a*, and b* were respec- crowave) prior to packing was very impor-
An additional reaction occurred, in both tively for garlic blanched with hot water 66.7, tant for the preparation of high-quality pick-
the pasteurized product and the control, from -2.9, 15.0 and by microwave, 64.7, -3.5, 15.0 led garlic. Several positive effects resulted
the beginning of storage: there was a grad- vs 80.6, -1.9, 21.8 for raw garlic. Firmness from this treatment: elimination of the pun-
ual appearance of a green coloration and the decreased by 59% (microwave) and 70% gent flavor by deactivation of the enzyme
pigment(s) responsible is unknown. In the (conventional), both compared to raw gar- allinase, elimination of green coloration or
pasteurized product this coloration became lic. Allicin was not detected after either formation of gas, and reduction of firmness.
evident a few hours after packing, while in blanching treatment. Pickled garlic, irrespective of preparation
the control it became clear after 2-3d. The Means of main effects of blanching, pres- method, was microbiologically stable during
intensity of the green coloration was consid- ervation treatment and storage time were storage, including that without a preserva-
erable after 9d storage in both products, al- compared on the chemical characteristics (Ta- tion treatment. Nevertheless, a preservation
though it was much stronger in the non-pas- ble 3) and quality parameters (Table 4) of treatment such as pasteurization would be
teurized control. This was shown by lower packed blanched garlic. Titratable acidity and advisable for safety reasons. All products, ex-
values of the color parameter a* (Table 2). concentrations of fructans, sucrose and cit- cept that using preservatives + refrigeration,
Later, the green coloration faded and both ric acid in microwave blanched garlic were resulted in hydrolysis of fructans during stor-
products became yellowish-brown, shown by higher than in water blanched garlic. The age.
increases in parameters a* and b*. color parameters -a* and b* were higher for
The firmness of the pasteurized product water blanched garlic, and firmness was
was considerably less than that of the raw greater for microwave blanched garlic. REFERENCES
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garlic, but that of the control barely differed Sucrose, fructose, glucose and citric acid nus Allium—Implications for the organic chemistry of
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kazawa, H., Suzuki, S., and Fujita, M. 1988. Liquid
samples treated with preservatives + refrig- chromatographic determination of alliin in garlic and
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Montaño, A., Sánchez, A.H., and Rejano, L. 1990. Rap-
perature during storage inhibited the hydrol- id quantitative analysis of headspace components of
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became more yellow), while L* decreased bacterial activity of phenolic and nonphenolic frac-
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shown). Packed blanched garlic did not de- StaSoft, Inc. 1996. STATISTICA for Windows (Comput-
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velop the green coloration that was a defi- Stoll, A. and Seebeck, E. 1951. Chemical investigations
ciency in fresh-pack garlic. However, in the on alliin, the specific principal of garlic. Adv. Enzy-
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microwave blanched product, the green col- Suzuki, M. and Cutcliffe, J.A. 1989. Fructans in onion
oration was observed at the ends (apparently bulbs in relation to storage life. Can. J. Plant Sci. 69:
1327-1333.
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(p<0.05) of firmness during storage time (Ta- Yu, T.-H. and Wu, C.-M. 1989. Stability of allicin in gar-
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Fig. 2—Effect of storage time of packed Ms received 2/10/97; revised 5/16/97; accepted 6/20/97.
blanched on garlic concentrations of fruc-
tans (a), fructose (b), and glucose (c). P, pas- CONCLUSIONS This work is part of a research project, AL194-0980-CO2-01,
supported by the Spanish Government through CICYT.
teurized; C, control; R, refrigerated. BLANCHING (CONVENTIONAL OR BY MI-

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