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The article presents a solution to blue garlic challenge. The pigment precursor was a 3,4-dimethylpyrrole derivative. It is thought to be formed by condensation of the amine group of Allium amino acids with the thial/thial S-oxide formed by - sigmatropic rearrangement of bis-1-propenyl thiosulfinate, in turn formed by the action of alliinase on 1-PeCSO (isoalliin). The thial/thial S-oxide is an intermediate in the formation of zwiebelanes, and is closely related in structure to (Z,Z)-d,1-2,3-dimethyl-1,4-butanedithial S,S'-dioxide, a compound isolated from onion preparations, which could play a role in forming the pigment precursor. The plant chemistry, however, is always wonderfully complex, so that other studies are not useless. The active compound that yielded blue color when combined with unheated onion juice was isolated from unheated garlic juice and was confirmed to be allicin, which derived from alliin by the action of alliinase.
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Solution to blue garlic challenge
Hervé This
#Springer-Verlag Berlin Heidelberg 2014
The winner of the blue garlic challenge (published in volume
406 issue 1) is:
Wai-Yin Lau, Department of Chemistry, University of Hong
Kong, Hong Kong, China
The award entitles the winner to select a Springer book of
her choice up to a value of 100.
Our Congratulations!
The explanation of the greeningor blueingof garlic
(Allium sativum L.) and pinkingof onion (Allium cepa L.)
given here will conclude with a new challenge of its own.
Color changes of Allium tissues have been studied by
several investigators since Joslyn, in 1958 [1]. In the 1960s,
the reactions involved in the formation of pink pigment in
onion purée were investigated [2] and a three-step reaction
scheme was proposed: (1) the formation of a colorless, ether-
soluble substance [the color developer (CD)] by the catalytic
action of alliinase (enzyme EC upon then-unknown
precursors in the fraction of neutral and basic amino acids; (2)
the formation of a colorless, ether-insoluble pigment precursor
(PP) from the color developer and an amino acid such as
glycine; (3) the formation of a pink pigment from the pigment
precursor PP and a naturally occurring carbonyl (NOC) sub-
stance such as formaldehyde. It was then assumed [3]thata
blue pigment-forming carbonylwas present in garlic, and
when reacted with the pigment precursor PP in place of NOC
from onion, a blue color would form.
Subsequently, it was found [4] that the blue pigment-forming
carbonylwas derived from isoalliin (a substrate for alliinase, a
minor precursor in garlic flavor, but the major precursor of onion
flavor), and the same reactions were responsible for greening
of garlic and of the mixture of garlic and onion.
But our understanding of the Allium chemistry advanced
significantly when Eric Block and his colleagues [58]intro-
duced methods for the study of Allium chemistry and pro-
duced a wealth of results based on such methods. Based on
their work, it was established (see Fig. 1) that the pigment
precursor was a 3,4-dimethylpyrrole derivative. It is thought
to be formed by condensation of the amine group of Allium
amino acids with the thial/thial S-oxide formed by [3]-
sigmatropic rearrangement of bis-1-propenyl thiosulfinate, in
turn formed by the action of alliinase on 1-PeCSO (isoalliin).
The thial/thial S-oxide is an intermediate in the formation of
zwiebelanes, and is closely related in structure to (Z,Z)-d,l-
2,3-dimethyl-1,4-butanedithial S,S'-dioxide, a compound iso-
lated from onion preparations, which could play a role in
forming the pigment precursor. A second key aspect to the
formation of colors in Allium preparation requires the inter-
mediacy of thioacrolein.
Using such information, a positive correlation between
thiosulfinate concentration and pink pigment formation was
first observed [9], and 1-propenyl-containing thiosulfinates
were confirmed to be the major color-developing compounds
[10]. Then in 2005, Bai et al. [11] studied the mechanisms of
the green color formation in Labagarlic, a preserve of garlic
including vinegar and sometimes sugar: both alliinase and
acetic acid are required for the color formation, and the de-
crease in the total thiosulfinates in garlic cloves is associated
with the pigment formation.
The plant chemistry, however, is always wonderfully com-
plex, so that other studies are not useless. Imai et al. [12]
established a model reaction system that comprised only well-
defined constituents and reported identifications of new
This article is the solution to the Analytical Challenge to be found at
H. This (*)
INRA/AgroParisTech, UMR 1145, Group of Molecular Gastronomy,
16 rue Claude Bernard, 75005 Paris, France
Anal Bioanal Chem (2014) 406:27432745
DOI 10.1007/s00216-014-7699-6
substances, which were involved in the pigment formation
along with various conditions that affected color development.
Addition of glycine suggested that proteins in the garlic juice
participated presumably in the pigment formation and that
they were less reactive than 75 % MeOH-soluble free amino
acids, such as glycine. The active compound that yielded blue
color when combined with unheated onion juice was isolated
from unheated garlic juice and was confirmed to be allicin,
which derived from alliin by the action of alliinase. It was also
confirmed that a vivid-blue color could be produced by using
a highly defined model reaction system comprising only iso-
lated alliin, pure glycine, and purified garlic alliinase. Later,
Imai et al. [13,14] isolated two pigment precursors and a
reddish-purple pigment (PUR-1) and determined their chem-
ical structures.
In 2007, Lee et al. [15] purified the green pigment, respon-
sible for greening in crushed garlic cloves, and they analyzed
it by liquid chromatographyelectrospray ionization mass
spectrometry (LC-ESI-MS), fast atom bombardment mass
spectrometry (FAB-MS), matrix-assisted laser desorption/
ionization time-of-flight mass spectrometry (MALDI-TOF-
MS), and nuclear magnetic resonance (NMR) spectroscopy.
The purified green pigment was highly polar and slightly
viscous, with a garlic odor, and easily turned to a yellow or
brown color with exposure to room temperature. The absorp-
tion spectrum in methanol showed a crude methanolic green
pigment-like profile with two absorbance maxima at 440 and
590 nm. Although complete isolation of the 411 Da com-
pound for proper structure elucidation was not achieved in
their experiments, the MS and NMR spectra of the 411 Da
green pigment suggested the ambiguous structural assignment
of one sulfur atom and odd number of nitrogen atoms, with
2530 carbon atoms, including aromatic ring. Therefore, it
was envisioned that the green pigment observed in crushed
garlic cloves was a new sulfur-containing nitrogenous water-
soluble compound differing significantly from all previously
reported green pigments in plants.
Obviously, the story is not fully over, but the worst is
when one tries to get this color, as I did. When I simply
put peeled garlic cloves in vinegar, the color did not
appear, whether I boiled the system or not, and even after
2 wk of maceration. According to Block [15], garlic heads
should be aged before immersion in vinegar in order for
the color to appear. Lets meet again in 4 months from
1. Joslyn MA, Peterson RG (1958) Food discoloration, reddening of
white onion bulb purees. J Agric Food Chem 6:754765
2. Shannon S, Yamaguchi M, Howard FD (1967) Reactions involved in
formation of a pink pigment in onion purees. J Agric Food Chem 15:
isoalliin, 1-PeCSO
alliin, 2-PeCSO
alliinase SS+
alliinase SS+
max 230
dipyrrole pigment max 570
tripyrrole pigment (proposed; blue-green)
steps N
Fig. 1 Formation of various
visible light absorbing
compounds from Allium bulbs
(from [15])
2744 H. This
3. Yamaguchi M, Shannon S, Howard FD, Joslyn MA (1965) Factors
affecting the formation of a pink pigment in purees of onion. Proc
Am Soc Hortic Sci 86:475483
4. Shannon S, Yamaguchi M, Howard FD (1967) Precursors involved
in the formation of pink pigments in onion purees. J Agric Food
Chem 15:423426
5. Bayer T, Wagner H, Block E, Grisoni S, Zhao SH, Neszmelyi A (1989)
Zwiebelanes: Novel 2,3-dimethyl-5,6-dithibicyclo[2.1.1]hexanes from
onion. J Am Chem Soc 111:30853086
6. Block E, Bayer T (1990) (Z,Z)-d,l-2,3-Dimethyl-1,4-butanedithial S,
S'-dioxide: A novel biologically active organosulfur compound from
onion. Formation of vic-disulfoxide in onion extracts. J Am Chem
Soc 112:45844585
7. Block E (1992) The organosulfur chemistry of the genus Allium
implications for organic sulfur chemistry. Angew Chem Int Edn 31:
8. Block E, Bayer T, Naganathan S, Zhao SH (1996) Allium chemistry:
Synthesis and sigmatropic rearrangements of alk(en)yl 1-propenyl
disulfide S-oxides from cut onion and garlic. J Am Chem Soc 118:
9. Lukes TM (1986) Factors governing the greening of garlic puree. J
Food Sci 51:15771582
10. Lee CH, Parkin KL (1998) Relationship between thiosulfinates and
pink discoloration in onion extracts, as influenced by pH. Food Chem
11. Bai B, Chen F, Wang Z, Liao X, Zhao G, Hu X (2006) Mechanism of
the greening color formation of Labagarlic, traditional homemade
Chinese food product. J. Agric. Food Chem. 2005, 53, 71037107.
Imai S.; Akita K.; Tomotake M.; Sawada, H.; Identification of Two
Novel Pigment Precursors and a Reddish-Purple Pigment Involved in
the Blue-Green Discoloration of Onion and Garlic. J Agric Food
Chem 54:843847
12. Imai S, Tsuge N, Tomotake M, Nagatome Y, Sawada H, Nagata T,
Kumagai H (2002) An onion enzyme that makes the eyes water.
Nature 419:685
13. Imai S, Akita K (2006) Tomotake, M; Shinsuke Imai, Kaori Akita,
Muneaki Tomotake, Hiroshi Sawada, Model Studies on Precursor
System Generating Blue Pigment in Onion and Garlic. J Agric Food
Chem 54:848852
14. Lee EJ, Cho JE, Kim JH, Lee SK (2007) Green pigment in crushed
garlic (Allium sativum L.) cloves: Purification and partial character-
ization. Food Chem 101:16771686
15. Block E (2009) Garlic and other Alliums: The Lore and the Science.
Royal Society of Chemistry, London
Solution to blue garlic challenge 2745
... Including to the physical development, increased density would also be observed from the hardness of the tissue. Heating causes a Maillard reaction which causes the white garlic to become yellow then light brown until dark brown in color [14]. The brown color comes from melanoidin which is formed during process. ...
Full-text available
Black garlic is popular as food supplement as well as herbal medicine due to its rich and beneficial chemical contents. However, the chemical changes during the process are rarely investigated. The purpose of this study was to determine the effect of heating time on sulfur-containing compound profiles and antioxidant activity during fermentation process at 60oC by solvent extraction followed by GC-MS analysis, as well as antioxidant activity, using DPPH method. The result indicated that black garlic underwent changes of sulfur-containing organic secondary metabolites, such as allicin and its derivative compounds. The process also increased antioxidant activity in black garlic
The new biologically active title compound (DL)-(III), which can be isolated from onion extracts, is stereoselectively synthesized by a simple oxidation of the known compound (I).
Reduction (LiAlH4) of propyl 1-propynyl sulfide (8) to (E)-1-propenyl propyl sulfide ((E)-10), C−S cleavage (Li/NH3) to lithium (E)-1-propenethiolate (Li (E)-11), and reaction with MeSO2Cl gives (E,E)-bis(1-propenyl) disulfide ((E,E)-2); i-Bu2AlH reduction of 8 to (Z)-10 and reaction with Li/NH3 and then MeSO2Cl gives (Z,Z)-2 via Li (Z)-11. Reaction of MeSO2SR (R = Me (12a), n-Pr (12b), CH2CHCH2 (12c), CHCHMe (12d)) with K (E)-11 gives (E,Z)-2 from (Z)-12d; Li (E,Z)-11 gives alkyl (E)- and (Z)-1-propenyl disulfides (MeCHCHSSR, R = Me (3a), n-Pr (3b), CH2CHCH2 (3c)) from 12a−c, respectively. Oxidation at −60 °C of (E,E)-, (Z,Z)-, and (E,Z)-2 gives (E)-1-propenesulfinothioic acid S-(E)-1-propenyl ester ((E,E)-13, (E,E)-MeCHCHS(O)SCHCHMe) from (E,E)-2, (Z,Z)-13 from (Z,Z)-2, and ca. 2:1 (E,Z)-13)/(Z,E)-13 from (E,Z)-2. Warming (Z,Z)-13 gives (±)-(1α,2α,3β,4α,5β)-2,3-dimethyl-5,6-dithiabicyclo[2.1.1]hexane 5-oxide (1a), endo-5-methyl-exo-6-methyl-2-oxa-3,7-dithiabicyclo[2.2.1]heptane (14a), and exo-5-methyl-endo-6-methyl-2-oxa-3,7-dithiabicyclo[2.2.1]heptane (14b). Warming (E,E)-13 gives 14a and 14b; (E,Z)-13/(Z,E)-13 gives (1α,2α,3α,4α,5β)-2,3-dimethyl-5,6-dithiabicyclo[2.1.1]hexane 5-oxide (1b), exo-5-methyl-exo-6-methyl-2-oxa-3,7-dithiabicyclo[2.2.1]heptane (14c), and endo-5-methyl-endo-6-methyl-2-oxa-3,7-dithiabicyclo[2.2.1]heptane (14d). Oxidation of 3a−c gives MeCHCHSS(O)R (4) and MeCHCHS(O)SR (5). At −60 °C, m-CPBA (2 equiv) converts (E,E)-2 into (Z,Z)-d,l-2,3-dimethyl-1,4-butanedithial 1,4-dioxide (26) while (Z,Z)-2 gives meso- and d,l-26. With NaIO4, 4/5 (R = Me) gives (E)- or (Z)-12a and MeCHCHSO2SMe (6a); with m-CPBA (Z)-MeS(O)CHMeCHS+O- (25a) forms. At 85 °C 2 gives 1:1 cis- and trans-2-mercapto-3,4-dimethyl-2,3-dihydrothiophene (29).
A Cook's tour is presented of the organosulfur chemistry of the genus Allium, as represented, inter alia, by garlic (Allium sativum L.) and onion (Allium cepa L.). We report on the biosynthesis of the S-alk(en)yl-L-cysteine S-oxides (aroma and flavor precursors) in intact plants and on how upon cutting or crushing the plants these precursors are cleaved by allinase enzymes, giving sulfenic acids—highly reactive organosulfur intermediates. In garlic, 2-propenesulfenic acid gives allicin, a thiosulfinate with antibiotic properties, while in onion 1-propenesulfenic acid rearranges to the sulfine (Z)-propanethial S-oxide, the lachrymatory factor (LF) of onion. Highlights of onion chemistry include the assignment of stereochemistry to the LF and determination of the mechanism of its dimerization; the isolation, characterization, and synthesis of thiosulfinates which most closely duplicate the taste and aroma of the freshly cut bulb, and additional unusual compounds such as zwiebelanes (dithiabicyclo[2.1.1]hexanes), a bis-sulfine (a 1,4-butanedithial S,S′-dioxide), antithrombotic and antiasthmatic cepaenes (α-sulfinyl disulfides), and vic-disulfoxides. Especially noteworthy in the chemistry of garlic are the discovery of ajoene, a potent antithrombotic agent from garlic, and the elucidation of the unique sequence of reactions that occur when diallyl disulfide, which is present in steam-distilled garlic oil, is heated. Reaction mechanisms under discussion include [3, 3]- and [2, 3]-sigma-tropic rearrangements involving sulfur (e.g. sulfoxide-accelerated thio- and dithio-Claisen rearrangements) and cycloadditions involving thiocarbonyl systems. In view of the culinary importance of alliaceous plants as well as the unique history of their use in folk medicine, this survey concludes with a discussion of the physiological activity of the components of these plants: cancer prevention, antimicrobial activity, insect and animal attractive/repulsive activity, olfactory–gustatory–lachrymatory properties, effect on lipid metabolism, platelet aggregation inhibitory activity and properties associated with ajoene. And naturally, comments about onion and garlic induced bad breath and heartburn may not be overlooked.
A preparation of alliinase (alliin alkylsulphenate lyase) from garlic and a system of pyridoxal and Cu +2 which simulates the catalytic action of alliinase caused the formation of pigment precursors in the amino acid fraction from onions. The precursors formed were an ether-soluble, ultraviolet-absorbing compound and one or more unidentified carbonyl compounds. Reactions for pigment formation are proposed.
The reactions involving and the conditions affecting the formation of pink pigments from precursors isolated from purees of white onion were investigated. A colorless ether-soluble precursor reacts with certain amino acids in onions to form a second colorless compound insoluble in ether. The latter compound then reacts with formaldehyde or naturally occurring carbonyls to form the pigment. The final pigment-forming reaction proceeds at about seven times the rate of the first reaction and has an optimum near pH 4.8. Pigment formation is inhibited by the sulfhydryl group of cysteine. Studies with isotopes indicate that amino acids and formaldehyde are incorporated in the pigment molecule. The rate and extent of pigment formation and the color of pigment formed were affected by the kinds of amino acids and carbonyls.
A water-soluble red pigment is formed in acidified macerates of white onion bulb tissue and in bruised, sliced onion tissue during dehydration. The resulting reddening in onion extracts preserved by addition of acetic acid or vinegar is objectionable and the pinking of dehydrated sliced white onions requires manual sorting. Pigment formation in purees is affected by variety, storage conditions, heat treatment, and acidity. Storage at 50°C., after maceration and before acidification, increases rate and intensity of reddening. Heating sliced onions in steam before maceration inhibits reddening. Reddening usually is accelerated in rate and increased in intensity by acidification with acetic acid to pH 3 to 3.5 and does not occur below 2.5 or above 5.5. The isolation and purification by solvent extraction, chromatographic separation on column and paper, its characterization by absorption spectroscopy, and other properties are reported. The pigment was a new nitrogenous water-soluble pigment which differed significantly from all previously reported red plant pigments. Knowledge of its structure and the factors influencing reddening will be useful in explaining the differences in susceptibility to reddening with variety and growing conditions, and in avoiding this by better selection of onion bulbs and control of the factors influencing reddening.
Storing garlic bulbs for a month at or above 23°C, prior to processing, prevented the production of a green pigment in the garlic puree. The amino acid S-(1-propenyl) cysteine sulfoxide was necessary for the development of the green color.