Food Control 83 (2018) 54e60

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Food Control
journal homepage: www.elsevier.com/locate/foodcont

Chemical characteristics of spice paprika of different origins

Helga Molna
Ko

r a, *, Eva
nya a, Zsolt Zala

n a, Ildiko

Bata-Vida

cs b, Rita To
€ mo
€ sko
€ zi-Farkas a,

s Sze
Andra
ka

cs , No
b
ra Ada
nyi a

a
u. 15, Hungary
Food Science Research Institute, NARIC, 1022, Budapest, Herman Otto
b
Agro-Environmental Research Institute, NARIC, 1022, Budapest, Herman Otto
u. 15, Hungary

a r t i c l e i n f o a b s t r a c t

Article history: Spice paprika powders from Bulgaria, China, Hungary, Peru, Serbia and Spain were examined to identify
Received 31 August 2016 the most important differences in their major characteristics and to attempt to find chemical compo-
Accepted 19 April 2017 nents revealing their origin. Bioactive components, contaminants, aroma and microbiological properties
Available online 21 April 2017
were determined: the amount of carotenoids, tocopherols, ascorbic acid and mycotoxins were deter-
mined by HPLC, flavoring properties by GC-MS, lead and cadmium content by atomic absorption spec-
Keywords:
trophotometry. Carotenoids were found at the highest concentration in samples from Peru and Spain and
Spice paprika powders
at the lowest, in Serbian samples. The concentrations of total tocopherol and ascorbic acid were found to
Carotenoids
Tocopherols
be greater in samples from Hungary than from China. Aroma components characteristic to the location
Ascorbic acid were detected in samples from each country of origin. The level of microbiological contamination was
Flavoring properties acceptable, while contaminants were found in Serbian, Spanish, Peruvian and Chinese samples. The
effect of sample origin was also investigated using near infrared spectroscopy.
© 2017 Elsevier Ltd. All rights reserved.

1. Introduction
In order to assess the composition of bioactive ingredients in
spice paprika products and to support the safety of the spice
product chains, a wide range of compositional examinations were
performed on commercial spice paprika samples. Samples from
several countries were analysed to find the most important dif-
ferences in their characteristic properties and to differentiate
among the samples by origin. Measurements were carried out on
spice paprika samples of Hungarian and foreign (Serbian, Spanish,
Chinese, Bulgarian, Peruvian) origin. Bioactive components (carot-
enoids, tocopherols, vitamin C), contaminants (mycotoxins, content
of lead and cadmium), volatile aroma compounds and microbio-
logical properties (mesophilic aerobic total count, mould and yeast
counts) were analysed, compared and sorted on the basis of near
infrared spectroscopy (NIRS).
Bioactive components are of utmost importance as carotenoids
(e.g. free capsanthins, capsanthin monoesters and diesters, b-
carotene) provide both the flavour and the colour of spice paprika
* Corresponding author.
E-mail addresses: h.molnar@cfri.hu (H. Molna

r), e.konya@cfri.hu (E.
nya), zs.
zalan@cfri.hu (Z. Zala
n), i.vidacs@cfri.hu (I. Bata-Vida
cs), r.farkas@cfri.hu
(R. To€ mo
€ sko
€zi-Farkas), a.szekacs@cfri.hu (A. Sze

ka
cs), n.adanyi@cfri.hu (N. Ad

(Palacios-Morillo, Jurado, Alc
azar, & Pablos, 2016). Moreover, the
flesh and the seeds of paprika contain considerable amounts of
different homologues of tocopherol, reactive precursor of vitamin
E, as well as L-ascorbic acid (vitamin C), the most important water-
soluble antioxidant in spice paprika. In turn, determination of the
origin of red paprika products is of high importance from both food
safety and commercial aspects, yet only a limited amount of
experimental data have been reported on this topic.
The “fingerprint” method combining the measurement of
strontium isotope ratios (87Sr/86Sr) and multi-element patterns
(Rb, Sr, Y, Zr, Mo, Cd, Ba, Pb, Th, U, Mg, Ca, Sc, Ti, Cr, Mn, Fe, Co, Ni, Cu,
Zn, As and rare earth elements) by using inductively coupled
plasma mass spectrometry (ICP-MS) and is suitable for the parallel
determination of bioavailable Sr and other elemental sources in soil
(Brunner, Katona, Stef
anka, & Prohaska, 2010). Furthermore, the
stable natural nitrogen isotope abundance (d15N) value is a proper
indicator of both cultivation types (fertilisation) and geographical
origin (Austin, Masahumi, & Yoshihiko, 2010).
In the case of examination of aroma extracts from spice paprika,
gas chromatographyeolfactometry can be applied for the evalua-
tion, identification of odour-active compounds and can be com-
bined with flavour dilution (FD) factors (Zimmermann &

Ko
Schieberle, 2000).
Contamination with mycotoxins (e. g. fumonisin B1, ochratoxin
anyi).

http://dx.doi.org/10.1016/j.foodcont.2017.04.028
0956-7135/© 2017 Elsevier Ltd. All rights reserved.

r et al. / Food Control 83 (2018) 54e60
H. Molna
A, sterigmatocystin) and pesticide residues (e. g. metalaxyl fungi-
cide) in spices can be detected by ultra-high performance liquid
chromatography (UHPLC) coupled to a high resolution Orbitrap
mass spectrometry (Orbitrap-HRMS) technique (Reinholds,
Pugajeva, & Bartkevics, 2016). NIRS combined with a modified
partial least squares algorithm as a regression method has been
found to be an alternative method for the analysis of aflatoxin B1
and total aflatoxins in spice paprika (Herna
ndez-Hierro, García-
Villanova, & Gonza
lez-Martín, 2008).
2. Material and methods
Overall, 53 paprika samples were collected, with product choice
to include different origins and quality. The selected sample group
contained spice paprika samples of Hungarian (22 from Szeged and
Kalocsa) and foreign (6 Serbian, 7 Spanish, 14 Chinese, 2 Bulgarian
and 2 Peruvian) origin. Among the paprika powders from Hungary,
there were samples of known origin and presumably mixed ones as
well.
2.1. Determination of carotenoids
Ground spice paprika (0.5 g) was extracted by adding 50 ml of a
mixture of 2:1:1 1,2-dichloroethane-acetone-methanol, mixed for
15 min, and filtered. After solvent evaporation under vacuum, the
residue was re-dissolved in 10 ml of a mixture of 55:35:10
isopropanol-acetonitrile-methanol. A Waters Alliance HPLC in-
strument (Model 2695 and Model 2996 Photodiode Array Detector,
470 nm) was used for the analysis of carotenoids. A Nucleosil C-18,
3 mm, 250
4.6 mm column with gradient elution (Daood & Biacs,
2005) was used. Data processing was performed by software
Empower (Waters Corp., Milford MA, USA).
2.2. Determination of ASTA
The colour of the spice paprika products was determined by the
standardised method of the American Spice Trade Association
(ASTA) according to the standard protocol MSZ 9681-5:2002: 0.1 g
of ground spice paprika sample was extracted by adding 100 ml of
acetone, placed in the dark for 2 h during which time it was mixed
every 10 min and the absorbance at 460 nm measured by a spec-
trophotometer. The ASTA colour unit was calculated by a formula
considering the measured absorbance, sample quantity and a
spectrophotometric factor.
2.3. Determination of tocopherols
From 0.5 g of ground paprika, tocopherols were extracted with
5 ml of 30% methanolic potassium hydroxide, 0.5 g ascorbic acid
and 20 ml methanol (35 min under reflux). After cooling, the
tocopherol fraction was extracted twice by 40 ml of n-hexane.
Hexane fractions were pooled and washed with distilled water
(3
), dried over anhydrous Na2SO4, evaporated, and re-dissolved in
5 ml of n-hexane. A Jasco HPLC (880-PU) instrument (Shimadzu C-
R6A Integrator and RF-535 Fluorescence HPLC Detector, 295 nm
and 320 nm) was used for analysis. Under normal phase chro-
matographic conditions, separation was performed on a Nucleosil-
100, 5 mm, 250
4.6 mm column with isocratic elution of 99.6:0.4
n-hexane-ethanol.
2.4. Determination of vitamin C
The sample (1 g) was mixed with 25 ml of 3% meta-phosphoric
acid solution, shaken mechanically for 15 min at 20  C, filtered, and
cleaned-up on a 0.45 mm filter (Chromafil A-45/25, Cellulose mixed
55
esters) before injection. An Agilent Technologies 1200 Series high
HPLC instrument (Quaternary Pump, Diode Array, Multiple Wave-
length Detector SL, 265 nm) was used. Separation of vitamin C was
performed on Nautilus Nucleosil C-18, 3 mm, 150
4.6 mm column
with gradient elution.
2.5. Determination of volatile aroma compounds
The aroma of spice paprika is generated by the simultaneous
presence of a number of volatile compounds including terpenes,
hydrocarbons, esters and carotenoid derivatives. Ground paprika
(1 g) was placed in a capped headspace vial (40 ml volume), heated
at 50  C for 30 min, then the SPME needle (100 mm PDMS coated
fused silica fibre, Supelco) were exposed at 50  C for 30 min. Vol-
atile aroma compounds were desorbed at 250  C in the GC injection
port and flushed into the GC column (Mazida, Salleh, & Osman,
2005). A HP 5890/II gas chromatograph (Hewlett Packard) equip-
ped with a 30 m
0.25 mm x 0.25 mm RH-5ms þ capillary GC
column and 5971 MS detector were used to analyse the volatile
compounds of the red paprika powders. Detection was performed
in the 35e350 mass range (Cso
ka, Amtmann, Nemes, & Kora
ny,
2013). Compound recognition was based on an MS identification
spectrum library (Wiley275.L spectrum library) after individual
background correction.
2.6. Determination of microbial contamination
Analysis of mesophilic aerobic total count was performed ac-
cording to the standard protocol MSZ EN ISO 4833: plate pouring
with Plate Count Agar (MERCK) and incubation at 30  C for 5 days.
Analysis of mould and yeast counts were performed according to
the standard protocol MSZ ISO 7954: plate pouring with Chlor-
amphenicol Glucose Agar (Biolab) and incubation at room tem-
perature for 5 days.
2.7. Determination of mycotoxins
A mixture of 10 g of ground paprika and 2 g of sodium chloride
was extracted with 40 ml of methanol-water (80:20), stirred for
30 min and filtered. Twenty ml of the filtrate was diluted with
70 ml of PBS buffer, and was filtered on a glass fibre filter
(Schleicher & Schuell GF 53). Eighteen ml of the filtrate was passed
over an affinity column (VICAM AflaTest WB), after that the column
was washed with 20 ml distilled water and the aflatoxin content
was eluted from the column with 1.5 ml of methanol and diluted
with 1.5 ml of distilled water. HPLC measurement was carried out
with a Shimadzu LC-10ADvp instrument (Shimadzu, Kyoto, Japan)
equipped with a Shimadzu RF-10AXL fluorescence detector
(360 nm, 440 nm). The analytical column used was a HiChrom
LiChrospher RP 18 with a 5 mm particle size (4.6 mm
150 mm).
The mobile phase was water:acetonitrile:methanol (ratio 60:20:20)
with a flow rate of 1 ml/min. The amount of the injected sample
was 100 ml.
2.8. Determination of lead and cadmium content
Lead and cadmium content of the paprika powders were
determined by atomic absorption spectrometry according to AOAC
official method 999.10. Samples were digested by cc. HNO3-H2O2
(8:2) in microwave oven; the solution was diluted with H2O.
2.9. Application of NIR measurement
Near infrared spectra of the paprika samples were collected on a
Bruker Tango NIR Spectrometer at wavelengths between 9100 and
56
r et al. / Food Control 83 (2018) 54e60
H. Molna

3952 cm1, using software Unscrambler 10.3 for the spectrum

evaluation.

3. Results and discussion

3.1. Carotenoid composition of the paprika samples

The amount of free capsanthins, capsanthin monoesters and

diesters and b-carotene present in a spice paprika sample corre-
lated well with its measured colour intensity. Products with high
ASTA value also showed outstanding total carotenoid content. In
the case of Peruvian spice paprika, the average ASTA colour unit
was found to be 140 ± 35.4, and the average total carotenoids
concentration was 3810 ± 1206 mg/g. This was the highest ASTA
colour value and total carotenoids content among the samples
examined. In the case of Serbian samples, the average ASTA colour
value was 101 ± 28.3, and the average total carotenoid concentra-
tion was found to be 2855 ± 744 mg/g, which was the lowest con-
Fig. 2. Tocopherol content of spice paprika samples from different countries of origin.
centration value among the samples analysed.
When studying the detailed composition of the different spice
paprika sample groups, the level of the different carotenoid sub- spice paprika samples from Hungary (544 ± 103 mg/g, Fig. 2). In
stances is characteristic (Fig. 1). The content of monoesters varied contrast, the Bulgarian samples had only 399 ± 4.48 m/g, while the
between 221 ± 0.3 and 345 ± 136.4 mg/g, and the concentration of Serbian, Spanish, Chinese and Peruvian samples contained
b-carotene varied between 233 ± 80.2 and 319 ± 98.8 mg/g. At the 473 ± 40.08 mg/g of total tocopherols. The main tocopherol
same time, the free capsanthins and capsanthin diesters showed component in paprika fruit flesh is a-tocopherol, the most biolog-
the opposite trend. The free capsanthin content was the highest ically active component of vitamin E. Paprika seeds mainly contain
(202 ± 69.9 and 226 ± 56.4 mg/g), while the concentration of cap- g-tocopherol that plays an important role in the stability of paprika
santhin diesters was the lowest (1064 ± 332.9 and 911 ± 233.9 mg/g) products (Biacs, Daood, Pavisa, & Hajdú, 1989). The average con-
in the samples from Hungary and Serbia. In contrast, the content of centration of a-tocopherol in the samples was found to be
free capsanthins was the lowest (73 ± 35 and 68 ± 28.4 mg/g), while 403 ± 51.10 mg/g, while the corresponding value of g-tocopherol
the concentration of capsanthin diesters was the highest was 53 ± 14.55 mg/g, indicating the ratio of paprika seeds in spice
(1252 ± 348 and 1491 ± 499.5 mg/g) in the samples from China and paprika products.
Peru. In turn, the calculated ratio of capsanthin diesters to free
capsanthins may indicate the origin of the samples: the capsanthin
diesters/free capsanthins ratio of the samples from Serbia, Hungary, 3.3. Vitamin C composition of the paprika samples
Spain, Bulgaria, China and Peru was found to be 4.0, 5.3, 8.1, 8.2, 17.1
and 22.0, respectively. This tendency is possibly related to the The level of vitamin C was highest in spice paprika samples from
climate of the country. Hungary (1353 ± 801 mg/g) and Serbia (1321 ± 814 mg/g) (Fig. 4). In
contrast, low vitamin C concentrations were found in the Spanish,
Chinese, Bulgarian, and Peruvian spice paprika samples (average
3.2. Tocopherol composition of the paprika samples
concentration: 162 ± 203.77 mg/g), perhaps due to differing spice
paprika processing techniques in these countries, but vitamin C
The highest concentrations of total tocopherols were detected
content of the samples also depends on the storage conditions and

Fig. 1. Carotenoid composition of spice paprika samples from different countries of Fig. 3. Microbiological characteristics of spice paprika samples from different coun-
origin. tries of origin.

r et al. / Food Control 83 (2018) 54e60
H. Molna
Fig. 4. Vitamin C content and mycotoxin contamination of spice paprika samples from
different countries of origin.
the age of the spice paprika products.
3.4. Volatile aroma compounds of the paprika samples
Volatile aroma compounds (like terpenes, hydrocarbons, esters,
carotenoid derivatives) occurring in 3 or more spice paprika sam-
ples were b-elemene, trans-caryophyllene, a-muurolene, dihy-
droactinidiolide, 2,2,4,6,6-pentamethyl-heptane and b-ionone
(Fig. 5). Acetic acid and neryl acetone were detected in each sample,
the percentage area of neryl acetone in the chromatogram was
remarkable in all examined spice paprika samples. Only the Hun-
garian spice paprika samples contained 3-hydroxy-2-butanone, 6-
methyl-5-hepten-2-one and 2,3-butanediol. Serbian samples con-
tained a-terpinolene, I-phellandrene and heptadecanoic acid.
Linalool was detected in the samples from Spain. In case of Chinese
samples, 1,3-butanediol and d3-carene were detected. 2-pentyl-
furan, octadecanoic acid methyl ester and (þ)-aromadendrene
Fig. 5. Aroma components in spice paprika samples from different countries of origin.
57
were found only in the Bulgarian samples. Only the Peruvian
samples contained geranyl acetone. In conclusion, aroma compo-
sition turned out to be a likely indicator of origin in case of the
examined paprika samples.
3.5. Microbial status of the paprika samples
In the European Union there is no specific requirement about
the examination of microbiological contamination of herbs and
spices. According to the requirements of the European Spice As-
sociation, Salmonella cannot occur in 25 g of samples and the
Escherichia coli number should be below 102 cfu/g. Hungarian
Government Decree (4/1998. (XI. 11.)) also requires the compulsory
examination of Salmonella (n ¼ 10, M ¼ 0/25 g) and Escherichia coli
(n ¼ 5, c ¼ 1, m ¼ 102, M ¼ 104), and recommends the examination
of total viable count (n ¼ 5, c ¼ 2, m ¼ 104, M ¼ 106), the number of
moulds and yeast (n ¼ 5, c ¼ 2, m ¼ 102, M ¼ 104) and the
Enterobacteriaceae number (n ¼ 5, c ¼ 2, m ¼ 102, M ¼ 103) as
additional tests. In our approach, assessment of the microbiological
status was not primarily aimed to ensure food safety, but rather to
obtain an overall picture on the microbial loads of paprika samples
of different geographical origins. Consequently, the microbial
analysis was focused on the total viable counts, as well as on mould
and yeast contamination, and not on the individual occurrence of
particular species (e.g. Salmonella spp. or Escherichia coli). In the
paprika powders the average number of total viable count was
3.74
106±1,501,545 cfu/g (Fig. 3). The average number of moulds
was 7.22
103±5267.54 cfu/g, while that of yeasts was
1.42
103±1746.04 cfu/g. There was no significant difference be-
tween paprika powders regarding the microbiological load and all
samples were appropriately safe.
3.6. Mycotoxin contamination of the paprika samples
Among mycotoxins, the greatest food safety risks in spice
paprika are attributed to the presence of aflatoxin B1 and OTA.
Therefore at present, legal limits apply for these mycotoxins in
spice paprika. The limit for OTA in the European Union is 15 mg/kg
according to Commission Regulation 105/2010/EU. Based on our
58
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H. Molna

Fig. 6. NIR spectra of spice paprika samples from different countries of origin (A) and the results of PCA of raw spectra (B, C, D) (red e Serbian, blue e Spanish, green e Hungarian,
pink e Chinese). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

r et al. / Food Control 83 (2018) 54e60
H. Molna
results, OTA was found in 2 out of 22 Hungarian spice paprika
samples, both were below the legal limit (0.77 ± 0.24 and
0.73 ± 0.04 mg/kg). In all, 19 out of 31 examined foreign spice
paprika samples were contaminated with OTA, from these the
contamination rate of a Spanish and a Chinese sample was over the
legal limit, 35.53 ± 0.4 and 19.57 ± 0.85 mg/kg, respectively. Serbian
spice paprika samples were free of OTA contamination. The legal
limit of total aflatoxins (B1þB2þG1þG2) in the European Union is
10 mg/kg according to Commission Regulation 165/2010/EU. Among
the 22 Hungarian samples 2 were contaminated with aflatoxin B1,
both of which were below the legal limit (4.33 ± 1.06 and
3.44 ± 1.11 mg/kg). Twelve of the 30 foreign spice paprika samples
were contaminated with aflatoxins, of which one Serbian and both
Peruvian samples were above the legal limit, 17.47 ± 7.72,
13.52 ± 6.76 and 10.79 ± 5.39 mg/kg, respectively. The concentration
of total aflatoxins was below the limit in all Spanish samples. Data
shows that the mycotoxin contamination rate of Hungarian paprika
powders was not detectable or below the limit.
In the case of mycotoxin contaminated spice paprika samples,
the concentration of vitamin C was very low or in several cases even
below the limit of detection (Fig. 4). For example in the 2 Peruvian
spice paprika samples the average concentration of OTA was
11 ± 4.16 mg/kg, and of total aflatoxins was 12.16 ± 1.93 mg/kg, while
the average level of vitamin C was only 36.8 ± 10.9 mg/g. This
suggests that these spice paprika samples were not very fresh or
that they were stored under improper storage conditions (high
humidity, high temperature). In contrast, in the Hungarian paprika
powders the average concentration of OTA was 0.07 ± 0.22 mg/kg,
and total aflatoxin content was 0.35 ± 1.15 mg/kg, while the average
level of vitamin C was 1353 ± 801 mg/g.
3.7. Lead and cadmium contamination of the paprika samples
Toxic heavy metals in spice paprika may also pose substantial
food safety risks. At present there are limits for these heavy metals
in spice paprika products. The legal limit for lead is 2000 mg/kg
according to the Hungarian Government Decree 40/2000. (XII. 20.).
Lead was detected in 1 out of 22 Hungarian spice paprika samples,
at a level of 334.2 ± 1.3 mg/kg i.e. below the legal limit. Among the
31 examined foreign spice paprika samples, 7 were contaminated
with lead. As for the 7 Spanish spice paprika samples, 3 were
contaminated with lead, and the average lead concentration was
310.4 ± 2.5 mg/kg, which was also below the legal limit. Three out of
14 Chinese spice paprika samples were contained with lead at an
average concentration of 399.6 ± 0.7 mg/kg, while 1 out of the 2
Bulgarian samples contained 54.1 ± 0.4 mg/kg. Serbian and Peruvian
paprika powders were free of lead contamination.
The legal limit for cadmium is 250 mg/kg according to Hungarian
Government Decree 40/2000. (XII. 20.). Seven Hungarian spice
paprika samples (7/22) were contaminated with cadmium, the
average concentration was 86.2 ± 2.2 mg/kg. In case of the Serbian
(2/5) and Peruvian samples (2/2), these values were 170.2 ± 1.1 and
254.9 ± 1.8 mg/kg, respectively. Cadmium contamination was not
detectable in the Spanish, Chinese and Bulgarian samples. In terms
of heavy metal contamination the examined paprika powders were
appropriately safe.
3.8. NIR evaluation of the paprika samples
As the first step of NIR spectra evaluation, principal component
analysis (PCA) was carried out to investigate the effect of the origin
of the samples. PCA was carried out both on the raw spectra
(Fig. 6A) and on their second derivatives to eliminate baseline shift.
Results of PCA of the raw spectra is shown on Fig. 6B, C, D, with the
different countries of origin indicated by different colours. A certain
59
degree of clustering by origin is seen for the two main principal
components (PC1 and PC2, Fig. 6D) particularly for samples origi-
nating from Spain and Hungary. While the Hungarian and Chinese
samples represent single groups, Spanish samples are clustered
into two, relatively distant sub-groups. The same effect could be
observed from the results of the PCA of the second derivatives of
the spectra.
4. Conclusions
In summary, Spanish and Peruvian samples were notable in
total carotenoids content. The ratio of capsanthin diesters to free
capsanthins of spice paprika samples are proposed as indicators of
origin, supposedly by being related to the climate of the given
country. In Hungarian spice paprika samples, the concentrations of
vitamin C and total tocopherols were high. Aroma components
characteristic to the geographical location were detected in each
spice paprika sample. The microbiological purity and heavy metal
contamination of each sample was acceptable. The features of the
terroir, genotype and the parameters of harvest, processing and
storage appear to influence the content of bioactive components,
aroma properties and microbiological characteristics of the sam-
ples. Vitamin C content of the samples may be indicative of the age
of the spice paprika product and of storage conditions. According to
the results of NIR evaluation of spice paprika samples, some clus-
tering among the samples occurred according the country of origin.
To summarize our data, no clear correlation between the examined
bioactive components and places of origin was identified. However,
according to the results of the volatile aroma compounds, their
composition may be an indicator of origin in case of paprika sam-
ples. Based on NIR evaluation, some clustering occurred also ac-
cording the country of origin.
Acknowledgements
This research was executed in the framework of the EU-project
SPICED (Grant Agreement: 312631) with the financial support from
the 7th Framework Programme of the European Union. This pub-
lication reflects the views only of the authors, and the European
Commission cannot be held responsible for any use, which may be
made of the information contained therein.
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