JFS: Sensory and Nutritive Qualities of Food

Rapid Colorimetric Method to Detect

Indole in Shrimp with Gas Chromatography
Mass Spectrometry Confirmation
S.L. SNELLINGS, N.E. TAKENAKA, Y. KIM-HAYES, AND D.W. MILLER

ABSTRACT: With increased public concern over the freshness and quality of seafood, more pressure is being applied
to the industry to provide better products in the market place. Simple, fast, and inexpensive tests are needed to assess
seafood quality. Seafood decomposition can be characterized by biogenic amine formation, such as the formation
of indole in shrimp. We have developed a rapid colorimetric method using 4-(dimethylamino)benzaldehyde method
that detects indole in decomposing shrimp. This method, as well as the confirmatory gas chromatography/mass
spectrometry (GC/MS) method, is centered on a simple toluene extraction technique that recovers indole with high
efficiency. Excellent agreement was obtained between the colorimetric and GC/MS quantitative results for shrimp
decomposition.
Keywords: colorimetry, indole, biogenic amines, 4-(dimethylamino) benzaldehyde, shrimp

Introduction ysis, wherein indole is derivatized into a variety of colored prod-

S EAFOOD CONTINUES TO GROW IN POPULARITY IN THE AMERICAN

diet, in part because of beneficial health effects and the wide-
spread availability of fresh and frozen products. There is increased
concern about the freshness of the seafood, the potential of bacte-
rial contamination, and the overall safety of seafood. Characteristic
compounds are formed as seafood spoils, and the presence of these
compounds can be used to ascertain freshness. A freshness indicator
for high-protein seafoods like shrimp and shellfish is indole, which
is a product of the bacterial decomposition of tryptophan (Trp). Vibri-
os, Pseudomonas, Shawenella, and Proteus are examples of bacteria
that produce indole, but others do as well (Harrow 1950; Nickelson II
and Finne 1984; Smith and others 1984; Gill 1990; Jensen and others
1995; Dart 1996; Gram and Huss 1996; Godinho and D’Souza 2000).
Accordingly, indole can be an indirect indication of the bacterial
load, hence the freshness, of shrimp. Fresh shrimp contain at most
0.01 ␮g indole/g shrimp, while decomposing shrimp contain much
higher levels (Chambers and Staruszkiewicz 1981). Globally, inde-
pendent research groups have proposed using indole as a fresh-
ness/quality index for shrimp (Boe and others 1982; Amodio and
others 1984; Niola and Valletrisco 1986; Thomas and Iyer 1994; Is-
abel and others 1996; Oetjen and Oehlenschlaeger 2000). Other
research groups have advocated the use of indole to establish time
and temperature abuse of fresh and frozen shrimp (Matches 1982;
Chang and others 1983; Smith and others 1984; Thomas and oth-
ers 1995; Mendes 2002).
There have been numerous analytical methods reported for the
determination of indole in shrimp. Recent publications include GC/
olfactory analysis (Friedrich and Acree 1998), headspace GC/MS
Sensory and Nutritive Qualities of Food
ucts. Derivatizing agents include phosphoric acid (Boctor 1972;
Patterson 1948), chloranil (Manzar and Fasihullah 1988), lignine
(Davidson and others 1973), electron acceptor molecules (Hutz-
inger 1969; Hutzinger and others 1972), sodium nitroprusside (Su-
gumaran and Vaidyanathan 1978), sodium nitrite-glycine-HCl (Boc-
tor and others 1978), halosuccinimides (Islam and others 1984),
haloacetamides (Gopal and others 1977), and most importantly, 4-
(dimethylamino) benzaldehyde (Duggan and Strausberger 1946;
Anthony and Street 1970; Ehmann 1977; Cheuk and Finne 1981;
Veveris and others 1988; Tantillo and others 1996).
Several versions of the 4-(dimethylamino)benzaldehyde
(DMAB) derivatization of indole in shrimp have been reported in
the literature, but each method involves tedious multi-step proce-
dures that extract indole from shrimp tissue. These time-consum-
ing extraction procedures include steam distillation (Patterson
1948), rotary-evaporation with successive extractions (Tantillo and
others 1996), and vacuum filtration with multiple solvent extrac-
tions (Cheuk and Finne 1981). Herein we introduce a very simple
colorimetric method that extracts 92% of the indole in decomposed
shrimp and quantitates the indole using DMAB. Colorimetric re-
sults were in excellent agreement with GC/MS analysis. This method
can be used to rapidly determine time-temperature abuse in
shrimp.
Materials and Methods
Chemicals and materials
Indole, 2-methylindole (2MI), 3-methylindole (skatole), 2,3-

(Harmon 2001), liquid chromatography (Mattivi and others 1999),
fluorometry (Jie and others 1996), and colorimetry (references given
below). Of these methods, only colorimetry offers instrumentation
(spectrophotometer, colorimeter) that is easy to use, small, portable,
and comparatively affordable. A simple colorimetric method for in-
dole could allow seafood regulators, inspectors, and wholesalers to
rapidly establish shrimp quality before it reaches the consumer.
There are several colorimetric methods available for indole anal-
dimethylindole (2,3MI), 4-(dimethylamino)benzaldehyde (DMAB),
and trichloroacetic acid (TCA) were purchased from Aldrich (Mil-
waukee, Wis., U.S.A.). Toluene, methanol (MeOH), and concentrat-
ed hydrochloric acid (HCl) were purchased from J.T. Baker (Phil-
lipsburg, N.J., U.S.A.). A 5 lb block of raw, unshelled gulf coast fresh
frozen shrimp was purchased from a local retailer (Little Rock, Ark.,
U.S.A.). The shrimp contained sodium bisulfite as a preservative,
and the quality of the shrimp was Class 1 at the beginning of the

1548 JOURNAL OF FOOD SCIENCE—Vol. 68, Nr. 4, 2003 © 2003 Institute of Food Technologists
Further reproduction prohibited without permission
Indole colorimetry of shrimp . . .
experiment and Class 3 at the end. Indole standards in toluene
were made in the 0.1 to 25 ␮g/mL concentration range for GC/MS
calibration; these standards were diluted by a factor of 25 for colo-
rimetric calibration.
Indole extraction procedure
Indole was extracted from shrimp by grinding 20 g shrimp with
50 mL toluene and 5 mL 5% TCA for 1 min. The purée was centri-
fuged with a Jouan CR 422 centrifuge for 30 min at 3500 rpm
(2534 × g) to separate the toluene from the shrimp pulp. The tolu-
ene layer was decanted and filtered through a 0.45 ␮m syringe filter
into a beaker containing anhydrous Na2SO4 (Aldrich).
Recovery experiments
The recovery of indole from shrimp was established by spiking
20 g shrimp with 10 ␮L of 10 mg/mL indole (MeOH as solvent) prior
to the blending of the shrimp. The extract was subjected to GC/MS
and colorimetric analyses, and the amount of indole recovered and
detected was determined. Unadulterated shrimp were analyzed as
a control in both methods. For recovery, 4 shrimp samples were
spiked with indole and analyzed in triplicate with colorimetry and
GC/MS.
Shrimp decomposition
Thawed, raw shrimp were decomposed at both room (18 °C) and
low (7 °C) temperatures, and the shrimp were sampled every 12 to
24 h for 5 d. The pH of selected samples was measured with DUAL
TINT Universal Indicator Paper ( J.T. Baker, Phillipsburg, N.J.,
U.S.A.). Colorimetric and GC/MS analyses of the samples were
performed in quadruplicate and duplicate, respectively, and then
the corresponding results were averaged.
Gas chromatography/mass spectrometry
Measurements were made in full scan mode with a Varian Sat-
urn 4D GC/MS ion trap spectrometer equipped with a DB5-MS
column (30 m length × 0.25 mm I.D. narrowbore × 0.25 ␮m film
thickness) purchased from J&W Scientific (Folsom, Calif., U.S.A.).
Column temperature: programmed at 13.4 °C/min from 120 to
240 °C, injection temperature: programmed at 200 °C/min from
110 to 250 °C, injection volume: 0.5 ␮L, ion-source temperature:
250 °C, ionizing voltage: 70 eV, linear flow: 1 mL/min through col-
umn. The internal standard used was 2MI; 30 ␮L of a 3 mg/mL stan-
dard in toluene was added to 10 mL of shrimp extract. Unadulter-
ated shrimp were used as a control in the analyses.
Colorimetry
The derivatizing reagent used in the colorimetric method con-
sisted of 1.25 g DMAB in 100 mL MeOH and 25.6 mL concentrated
HCl (Snell and Snell 1967). In the procedure, 400 ␮L shrimp extract
was diluted to 10 mL with toluene, after which 2 mL was vortexed for
15 min with 2 mL derivatizing reagent. The resulting mixture was
centrifuged for 6 min at 3500 rpm to separate the MeOH and tolu-
ene layers. The absorbance of the MeOH (bottom) layer was mea-
sured with a Hach DR/2000 spectrophotometer at 567 nm. Unadul-
terated shrimp were used as the blank for the absorbance
measurements. A Cary 50 Conc UV/VIS spectrophotometer was
used in experiments involving multiple wavelength analysis of
derivatized samples.
Results and Discussion
GC/MS calibration and recovery
The Saturn 4D GC/MS was used to monitor the most significant
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ion for indole (M+, m/z 117) and the internal standard 2MI (M–1
ion, m/z 130). A qualifier ion (M–HCN ion, m/z 103) (Porter 1985)
was monitored for 2MI so peak purity could be established. Indole
and 2MI had retention times of 5.35 and 6.37 min, respectively. The
response of the Saturn 4D GC/MS was calibrated against 7 indole
standards ranging from 0.1 to 25 ␮g/mL indole using 2MI as the
internal standard. The GC/MS response as a function of indole
concentration was linear (data not shown), and Eq. 1 gives the
best-fit line (R2 = 0.996) in terms of indole (I), 2MI, and concentra-
tion (C). The lower limit of detection for the method is 100 ppb (0.1
␮g/mL); the upper limit of detection was not determined because
it far exceeded the amount of indole in fully decomposed shrimp.
Eq. 1
Figure 1 shows a chromatogram of a shrimp sample spiked with
100 ␮g indole. Using the extraction procedure detailed in the exper-
imental section, an average of 92.3 ± 4.3 ␮g indole was recovered
from shrimp spiked with indole before the blending of the shrimp.
The recovery of indole was accounted for in all calculations. The
high extraction efficiency shows the demonstrated solubility of
indole in toluene rather than the shrimp pulp/TCA/MeOH mixture.
Toluene readily extracts indole because of the aromaticity and sim-
ilarity of the molecules.
Colorimetric calibration and derivatization
The absorbance response of the Hach DR/2000 spectrophotom-
eter was calibrated against 2 concentration ranges of indole. The
first range tested matched that used for GC/MS calibration (0.1 to
25 ␮g/mL) and the results are shown in Figure 2. The absorbance
relationship to the indole concentration is clearly nonlinear
(R2 = 0.732) due the saturation of the detector by the deeply col-
ored solutions. Similar reports of non-linearity in calibration curves
using analogous concentration ranges have appeared in the liter-
ature (Cheuk and Finne 1981; Tantillo and other 1996), although
the nonlinearity is not as severe as that just presented.
Sensory and Nutritive Qualities of Food
Figure 1—GC/MS chromatogram for a shrimp sample
spiked with 100 ␮g indole: (a) total ion chromatogram, (b)
m/z 117 ion chromatogram for indole, and (c) m/z 130 ion
chromatogram for 2-methylindole. Indole and 2-
methylindole have retention times of 5.35 and 6.37 min,
respectively.
Vol. 68, Nr. 4, 2003—JOURNAL OF FOOD SCIENCE 1549
 Indole colorimetry of shrimp . . .

It was apparent in the first calibration that the 3 lowest concen- Table 1—Comparison of GC/MS and Colorimetric results
trations (0.1, 0.5, 1 ␮g/mL) formed a straight line (R2 = 0.998). Ac- for the decomposition of shrimp over 5 d at 18 °C
cordingly, the original 7 indole standards (0.1 to 25 ␮g/mL) were Temperature (°C) Time (d) GC/MS a Colorimetry a
diluted by a factor of 25 so that the maximum concentration tested 7 0 N/Db N/D
was 1 ␮g/mL. The diluted standards gave a linear absorbance re- 2 N/D N/D
sponse (data not shown), and Eq. 2 gives the best-fit line 2.5 N/D N/D
(R2 = 0.960) in terms of diluted concentration (CD) and absorbance 3 N/D N/D
5 N/D N/D
(A). Equation 3 relates the diluted concentration (CD) to the origi-
nal concentration (C) of the standard or extract. The detection limits 18 0 N/D N/D
2 N/D N/D
for this colorimetric method are 0.002 and 1 ␮g/mL.
2.5c 2.2 ± 0.7 2.1 ± 0.9
3c 12.5 ± 0.0 12.1 ± 0.2
Eq. 2 5c 26.8 ± 6.2 27.0 ± 6.5
a Concentration in the units of ␮g indole/g shrimp, corrected for recovery
b N/D: Indole levels not detectable
c Shrimp sample Class 3 in quality
Eq. 3

Using the colorimetric procedure, an average of 68.7 ± 2.2 ␮g in-

dole was detected from shrimp spiked with 100 ␮g indole. After
extraction into toluene, 92.3% of the indole present in the shrimp is
available for derivatization. From this, 68.7% is derivatized by
DMAB within 15 min leaving 23.6% of the indole underivatized in
the toluene layer. The 15 min reaction time was chosen as a balance
between analytical quantitation (and reproducibility) and speed of
the method. The absorbance response of the indole derivative is
linear with concentration, as shown in the above equations, indicat-
ing that the 15 min reaction time is sufficient for quantification. The
68.7% derivatization of indole by DMAB was accounted for in all
calculations.
Shrimp decomposition
The colorimetric method was used to determine the indole con-
tent of shrimp that were decomposed over a period of 5 d at both
low (7 °C) and room (18 °C) temperatures. Table 1 summarizes the
GC/MS and colorimetric results for both studies, while Figure 3
gives a graphical representation of the data. Table 1 shows that the
amount of indole detected by colorimetry is in excellent agreement
with that from GC/MS, with less than 1% difference between the re-
sults. The outstanding agreement between the 2 methods points
out an important aspect of the colorimetric method: the dilution of
Sensory and Nutritive Qualities of Food
the standards and extracts does not add additional error to the ab-
sorption data.
For low temperature decomposition, neither colorimetry nor GC/
MS detected indole in the shrimp throughout the 5 d study. The low
temperatures most likely the suppressed bacterial action in the
shrimp tissue (Fieger and Novak 1961), so no degradation of pro-
teins occurred and no free TRP residues were present (see next
section). Our results are similar to those from the literature (Match-
es 1982; Chang and others 1983; Amodio and others 1984; Thomas
and others 1995; Goswami and others 2001; Mendes and others
2002).
For the room temperature study, 2 ␮g indole/g shrimp was de-
tected by both analytical methods after 2.5 d. The decomposed
shrimp was classified as Class 3 by a trained organoleptologist (Par-
sons 2000). Class 3 is defined by the FDA to be severely decom-
posed and not fit for human consumption, while Class 1 is accept-
able for human consumption. The level of indole rose to a high of
27 ␮g indole/g shrimp in the shrimp at d 5. Similar reports of indole
formation during room temperature decomposition have appeared
in the literature (Duggan and Strausburger 1946; Barry and others
1956; Matches 1982; Chang and others 1983; Isabel and others
1996; Oetjen and Oehlenschlager 2000; Mendes and others 2002),
but our measured levels of indole in decomposed shrimp are high-

Figure 3—Indole formation as a function of decomposition

Figure 2—Nonlinear calibration of the colorimetric test time at both room (18 °C) and low (7 °C) temperatures. GC/
using 0.1 to 25 ␮g/mL indole standards (R2 = 0.732) MS and colorimetric data are corrected for recovery.

1550 JOURNAL OF FOOD SCIENCE—Vol. 68, Nr. 4, 2003 JFS is available in searchable form at www.ift.org
Indole colorimetry of shrimp . . .

er. This may result from the toluene extraction efficiency compared Table 2—Absorbance at 567 nm of several derivatized in-
to that of steam distillation or methanol extraction. doles
Compound Absorbance Molar absorptivity Color
Production of indole DMAB 0.000 67 Pale yellow
Indole is produced enzymatically by the metabolism of proteins Indole 0.116 27094 Light pink
by bacteria. Specifically, proteolytic bacteria hydrolyze proteins 2-Methylindole 0.081 18912 Pink
into free amino acids using proteinase and peptidase enzymes. Skatole 0.006 1475 Pale yellowa
2,3-Dimethylindole 0.000 67a Pale yellowa
Free Trp residues then undergo decarboxylation and deamination
Shrimp Extract 0.062 27094 b Pale pink
reactions, which are catalyzed by the bacterial enzyme tryptopha- a Same as the DMAB blank
nase, to produce indole and indole-containing molecules. A simple b Same as indole
multi-step pathway for Trp degradation is illustrated in Figure 4
(Vederas and others 1978; Hammond and Carlson 1980); indole
and skatole are the final products in this pathway. However, path-
ways more complex than that in Figure 4 have been reported (Har- such as Pseudomonas, Shawenella, Vibrios, and Proteus function
row 1950; Yokoyama and Carlson 1974; Marklová 1999). well under the conditions of temperature and pH (Jacobs and Ger-
The GC/MS and colorimetric data for shrimp decomposition stein 1960; Fieger and Novak 1961; Smith and others 1984; Gram
suggest that indole is the only indolic decomposition product in the and Huss 1996; Godinho and D’Souza 2000).
shrimp extract. Similar reports of indole formation with no detect-
able intermediates have appeared in the literature (Thimann 1955; Derivatization of indoles by DMAB
Jensen and others 1995). Jensen and others (1995) reported that DMAB is a very sensitive reagent that is used to detect trace
indole (85%) and skatole (15%) are detectable decomposition prod- amounts of indole, and a possible reaction mechanism is given in
ucts in entire male pig feces at pH 8. Thus, the conversion of Trp Figure 6 (Veveris and others 1988). The absence of skatole makes
into indole is the dominant branch of the degradation pathway the direct colorimetric analysis of indole simpler, but the coderiva-
(Figure 4), in which Trp breaks down into indole, pyruvate, and tization of skatole is not a deterrent to colorimetry because both
ammonia by the mechanism given in Figure 5 (Thimann 1955; Ved- molecules are a measure of quality.
eras and others 1978). The reactivity of DMAB towards indole compared with other
We measured the pH of the least (d 0) and most (d 5) decom- molecules containing the indole moiety was established by observ-
posed shrimp samples and both had a pH of 9 or higher. This results ing the color and absorbance of the derivatized products. Indole,
from the high protein and low carbohydrate content in the shrimp 2MI, skatole, and 2,3MI were reacted with DMAB under the same
tissue (Gram and Huss 1996); consequently, proteins and amino reaction conditions. Each indole had a concentration of 0.5 ␮g/mL
acids are broken down in large quantities during decomposition, after a 25-fold dilution from 12.5 ␮g/mL, which is a concentration of
producing significant amounts of ammonia (Thimann 1955). The indole higher than that found in the most rotten shrimp (d 5). The
pH 9 environment seems sufficient to inhibit the skatole branch of absorbance response of the derivatized indoles is shown in Figure
the pathway, allowing indole to be produced exclusively. Bacteria 7, and the data clearly indicates that indole and 2MI reacted with
DMAB to form colored products that strongly absorbed light be-
tween 500 and 600 nm. Derivatized skatole produced a negligible
absorbance response relative to the background at this concentra-
tion, as did 2,3MI. Table 2 summarizes the absorbance and molar
absorptivities (e) of each derivatized indole at 567 nm. Included in

Sensory and Nutritive Qualities of Food

Figure 4—Decomposition pathways of tryptophan into in- Figure 5—Breakdown of Trp into indole, pyruvate, and NH3
dole and skatole (Vederas and others 1978; Hammond and at alkaline pH (Thimann 1955; Vederas and others 1978).
Carlson 1980) Final products are underlined.

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 Indole colorimetry of shrimp . . .
Figure 7 as a comparison is the absorbance response of derivatized
shrimp extract (d 3); the identical shape of the shrimp extract and
indole curves is further evidence that indole is produced exclusively
during decomposition.
The formation of a colored derivative is dependent on the elec-
tronic nature of the indolic compound. The derivatization of indole
by DMAB involves the formation of an adduct in which the carbo-
nyl group of DMAB coordinates with the 2,3-double bond in indole.
This complex forms a covalent bond with either the 2- or 3- position
carbon, leaving a positive charge at the other carbon. According to
Millar and Springall (1960), electrophilic aromatic derivatization of
indole favors the 3-position because the incipient positive charge
can be delocalized into the aromatic system, as shown in Figure 6.
In the case of 2-position derivatization, the reaction product will
not be completely aromatic, so extended conjugation does not ex-
ist. Although skatole did not yield a colored product in the experi-
ment described above, skatole will form a purple-colored product
for undiluted concentrations of 0.5 ␮g/mL and above. For 2,3MI,
methyl substituents at the 2- and 3-positions prevent reactions with
DMAB. The color intensity, or degree of the delocalization of the
positive charge into the aromatic system, is exhibited in the high ⑀
values for indole and 2MI and the low ⑀ values for skatole and 2,3MI
(Table 2).
Figure 6—Mechanism for the derivatization of indole by
DMAB in an acidic medium (Veveris and others 1988)
Sensory and Nutritive Qualities of Food
Figure 7—Absorbance of derivatized indoles and shrimp
extract at wavelengths between 500 and 600 nm
1552 JOURNAL OF FOOD SCIENCE—Vol. 68, Nr. 4, 2003
Interferences
The coderivatization and detection of Trp residues in shrimp
proteins in the colorimetric method is undesirable because of the
corresponding artificial increase in absorbance. As prevention, TCA
is blended with the shrimp to precipitate proteins so that they are
not exposed to the DMAB reagent (Anthony and Street 1970; Cheuk
and Finne 1981). Free tryptophan residues present in the blended
shrimp are zwitterionic molecules between pK 3.1 and 8.0 (the car-
boxyl and amino groups are charged), while the residues are ionic
at pK > 8.0 (charged carboxyl group and neutral amino group) or
pK < 3.1 (charged amino group and neutral carboxyl group) (Stryer
1995). Therefore, Trp will not partition into toluene.
Shrimp carotenoids are extracted from the tissue by toluene,
which assumes the amber color of the compounds. The carotenoids
do not interfere with the colorimetric method because they do not
partition to the methanol layer, where the spectrophotometric
measurements are made. The toluene layer remains amber-col-
ored after the addition of the colorimetric DMAB reagent to the
extract. For GC/MS, the carotenoids do not chromatograph under
these conditions. Preservatives and packaging aids like sulfites,
phosphates, and lactic acid slow decomposition in shrimp, but
they do not interfere with indole detection because they are water-
soluble and do not partition into toluene.
Conclusion
A VERY SIMPLE EXTRACTION AND COLORIMETRIC PROCEDURE FOR
the detection of indole in a shrimp sample was described. The
revised colorimetric method increases the linearity of the absor-
bance response as a function of indole concentration, yielding ex-
cellent quantitative success when compared to GC/MS. Overall, the
colorimetric method described in this paper is both sensitive and
discriminating for indole in decomposing shrimp, but it is not in-
tended to replace sensory analysis. This method can confirm de-
composition that has occurred before and during processing, which
can remove and/or eliminate other markers of decomposition and
temperature abuse like ammonia and trimethylamine (Ottes
1981).
Improvements to the GC/MS method may be necessary if a
food sample decomposes below pH 8. At these pH values, skatole
can form measurable quantities (Jensen and others 1995). Current-
ly, 2MI is used as the internal standard, but 2MI and skatole have
nearly identical retention times on the DB5-MS column. By chang-
ing the internal standard from 2MI to 2,3MI, the indole and skatole
peaks can be separated and quantitated by GC/MS. Only a slight
change to the procedure is necessary for the colorimetric method.
If indole and skatole are both present in a sample, the total absor-
bance of the diluted extract has to be measured at the wavelength
of maximum absorption for derivatized indole (567 nm) and ska-
tole (582 nm). Solving the 2 simultaneous equations yields the con-
centration of indole and skatole. Another benefit of using 2,3MI as
the internal standard for GC/MS is that it does not react with
DMAB. This means that the internal standard could be spiked into
the shrimp tissue before analysis, thus providing recovery and
quantitation in 1 experiment.
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MS 20020670 Submitted 11/22/02, Revised 12/24/02, Accepted 1/21/03, Received
1/30/03
We are grateful to J. Wilkes for obtaining the shrimp used in this study, W. Parsons for the
organoleptic analysis of selected shrimp samples, and S. Laney-Sheehan for her assistance
with literature references. R. Cecotti is acknowledged for the translation of Italian articles
into English. All are from the National Center for Toxicological Research in Jefferson, AR.
This research was supported in part by an appointment of S.L.S. to the Research Participa-
tion Program at the National Center for Toxicological Research administered by the Oak
Ridge Institute for Science and Education through an interagency agreement between the
U.S. Dept. of Energy and the U.S. Food and Drug Administration.
Authors Snellings and Miller are with the Division of Chemistry, National
Center for Toxicological Research, Jefferson Laboratories of the FDA, 3900
NCTR Road, Jefferson, AR, 72079. Author Takenaka is with the Brown &
Williamson Tobacco Corp., 2600 Weaver Rd., Macon, GA 31217. Author Kim-
Hayes is with Magellan Laboratories Inc., 160 Magellan Lab Court.
Morrisville, NC 27560. Direct inquiries to author Miller (E-mail:
dmiller@nctr.fda.gov).
Sensory and Nutritive Qualities of Food
Vol. 68, Nr. 4, 2003—JOURNAL OF FOOD SCIENCE 1553