Journal of Liquid Chromatography & Related

Technologies

ISSN: 1082-6076 (Print) 1520-572X (Online) Journal homepage: http://www.tandfonline.com/loi/ljlc20

A review of thin layer chromatography methods

for determination of authenticity of foods and
dietary supplements

Joseph Sherma & Fred Rabel

To cite this article: Joseph Sherma & Fred Rabel (2018): A review of thin layer chromatography
methods for determination of authenticity of foods and dietary supplements, Journal of Liquid
Chromatography & Related Technologies, DOI: 10.1080/10826076.2018.1505637

To link to this article: https://doi.org/10.1080/10826076.2018.1505637

Published online: 08 Oct 2018.

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JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES
https://doi.org/10.1080/10826076.2018.1505637

A review of thin layer chromatography methods for determination of

authenticity of foods and dietary supplements
Joseph Shermaa and Fred Rabelb
a
Department of Chemistry, Lafayette College, Easton, PA, USA; bChromHELP, LLC, Woodbury, NJ, USA

ABSTRACT ARTICLE HISTORY

This paper reviews advances in qualitative and quantitative thin layer chromatography analytical Received 17 July 2018
methods published from 2000–2018 for assessing quality, adulteration, and authenticity of foods, Accepted 25 July 2018
beverages, and dietary supplements. Samples covered include animal and butter fats, oils, spices,
KEYWORDS
flavorings, grains, honeys, soft drinks and juices, tea, sweeteners, wine and other alcoholic bever-
Thin layer chromatography;
ages, and vegetables, which are analyzed for a wide variety of characteristic analytes. It is sug- densitometry; food
gested that TLC methods of the kind cited will continue to be used to complement the LC-MS/MS authentication; food
based peptide analysis methods that are being developed for food allergen detection and food adulteration; quality control;
fraud testing. food fraud; dietary
supplement; review
GRAPHICAL ABSTRACT

Introduction contaminants, and decomposition products for the deter-

[1] mination of alkaloids; amines, amides, and amino acids;
A previous review of thin layer chromatography (TLC) in
food and agricultural analysis included references up to anthocyanidines; anthraquinones; antibiotics; carbohydrates;
1999 on food composition, intentional additives, adulterants, flavonoids; imidazoles; lipids; pesticides, phenols; pigments

CONTACT Fred Rabel frabel@comcast.net ChromHELP, LLC, 136 Progress Ave, Woodbury, NJ 08096, USA.
Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/ljlc.
ß 2018 Taylor & Francis
2 J. SHERMA AND F. RABEL
and dyes; saponins; sulfonamides; toxins; vitamins; and
other miscellaneous compound types. A list of advantages of
TLC compared to analyses by high performance column
liquid chromatography (HPLC) and gas chromatography
(GC), including simplicity, minimum required sample prep-
aration, less solvent consumption, and possibility of analyz-
ing multiple samples side by side in less time, are also
provided in the earlier review. This review updates coverage
of the most important TLC analysis methods reported in the
literature up to mid 2018 for assessment of food (including
beverages) and dietary supplement authenticity and adulter-
ation in order to determine misrepresented ingredients.
Food fraud motivated by intentional contamination for eco-
nomic gain by manufacturers and potential terrorism with
chemical or biological agents against food supplies causing
toxic effects on consumer health is a critical concern at this
time, and TLC is an important analytical tool for quality
control (QC) and safety evaluation in its detection.
Techniques of TLC food analysis
This section will describe the general techniques that have
been applied for food analysis studies by TLC, and the next
sections will present examples of selected applications with
specific experimental details provided in each paper.
Foods are usually extracted prior to TLC using an appro-
priate solvent with traditional techniques such as shake,
homogenization, Soxhlet, or ultrasound assisted extraction
(UAE). Liquid samples such as wines can often be applied
as packaged with no sample preparation. Solid phase extrac-
tion (SPE) and supercritical fluid extraction (SFE) are more
modern techniques applied for sample preparation prior to
TLC analysis. Solvent mixtures described for sample prepar-
ation in this article are given in volume proportions, v/v,
unless otherwise noted.
Sample and standard solutions are applied to the plate as
spots manually with a micropipet or syringe or automatic-
ally as bands or spots with a commercial instrument.
Stationary phases reported were usually Merck 20
20 or
10
20 cm glass, aluminum, or plastic backed TLC or high
performance TLC (HPTLC) normal phase silica gel 60 plates
(60 nm pore size) with an organic binder and with or with-
out a fluorescent phosphor [plates with the designation F or
F254 or UV or UV254 contain a phosphor that emits green
light when irradiated with 254 nm ultraviolet (UV) light in
order to facilitate fluorescence quenching detection of com-
pounds that absorb this radiation and appear as dark zones].
Plates designated with a G contain gypsum inorganic binder.
Reversed phase (RP) octyldecylsilyl (C18 or RP18) chem-
ically bonded silica gel, amino (NH2) bonded silica gel, alu-
minum oxide, cellulose, and impregnated silica gel (e.g.,
with caffeine or silver nitrate) have been used when the
necessary selectivity and efficiency for the food constituent
separation were not provided by plain silica gel. Plated des-
ignated W are wettable with water. Thicker layers
(0.5–2 mm) are often used in preparative TLC (PTLC) for
purification of separated compounds by scraping layer
fractions, elution, and filtration to remove stationary phase
particles prior to further analysis. Unless otherwise stated,
plates used in applications were from Merck.
Plate development is usually done at laboratory tempera-
ture in a large volume, mobile phase vapor saturated N-
chamber in the one dimensional ascending or horizontal
mode. Forced flow overpressured layer chromatography
(OPLC) and automated multiple development (AMD) with
a CAMAG chamber and isocratic or gradient elution have
also been reported.
The method used for choosing the mobile phase is sel-
dom given in research papers, but it is almost always based
on a guided trial and error approach employing the analyst’s
personal knowledge of the analyte, layer, and mobile phase
properties and literature searching for previous systems
applied for the required separation. Mobile phase composi-
tions given in this article are in v/v proportions unless
otherwise noted, “ammonia” stands for concentrated ammo-
nium hydroxide, 28–30% NH3; “acetic acid” for glacial acetic
acid; “hexane” for n-hexane, and “water” is deionized
or distilled.
If the analyte is not naturally colored or does not quench
fluorescence on an F plate, detection of its separated zones
is achieved by viewing plates in daylight or under 254 or
366 nm UV light after applying a derivatization (detection)
reagent by spraying or dipping followed by heating if neces-
sary. Chromatogram quantification is by slit scanning densi-
tometry or image analysis.
HPTLC is usually carried out in literature papers using
CAMAG instrumentation, e.g., Linomat 4 or 5 or Automatic
TLC Sampler 4 (ATS4) band or spot applicator; Twin
Trough Chamber (TTC), Horizontal Developing Chamber,
or Automatic Developing Chamber (ADC2); Chromatogram
Immersion Device for applying detection dip reagents;
Digistore 2 Documentation System or TLC Visualizer for
chromatogram documentation; TLC Scanner 2, 3, or 4 for
densitometric quantification or Videoscan Digital Image
Evaluation Software for image analysis; and TLC-MS
Interface. CAMAG instruments are usually controlled by
winCATS or visionCATS software. Use of the Desaga appli-
cator AS-30, automatic separation chamber DC-MAT, scan-
ner CD-60, and ProQuant software is also specified in some
papers, but much less often. Validation of quantitation
methods is usually carried out by ICH (International
Conference on Harmonization) guidelines for specificity, lin-
earity, limit of detection (LOD), limit of quantification
(LOQ), precision, accuracy, robustness, and ruggedness.
Applications of TLC in food analysis
References are arranged in chronological order in the sub-
sections below.
Animal fat analysis
Methyl sterols were analyzed as chemical descriptors to
authenticate the quality of pig fat, which makes the animal’s
food products most appreciated by consumers due to their

aroma, taste, and texture. The determination of 4-methyl-
sterols and 4,4-dimethylsterols in 47 samples of subcutane-
ous fat from pigs reared on different fattening systems
comprised lipid extraction by melting the fat in a microwave
oven, GC-mass spectrometry (MS) identification, and quan-
tification by capillary column GC-FID (flame ionization
detector) with an SPB-5 column and hydrogen carrier gas.
For PTLC, the complete unsaponifiable fraction was dis-
solved in chloroform, the solution was spotted on a silica
gel 60 plate impregnated with potassium hydroxide, the
plate was developed twice with hexane-diethyl ether (65:35),
and then it was sprayed with 2,7-dichlorofluorescein to visu-
alize pink methylsterol bands under UV light. The band was
scraped off, methylsterols were dissolved in chloroform-
diethyl ether (1:1), and the solution obtained was filtered
and used for GC-FID and GC-MS analyses. Six pattern rec-
ognition techniques were applied to differentiate between
the fattening systems.[2]
Butter fat analysis
Ten-20% adulterant beef tallow in cow ghee (butter fat), an
important dairy product in India, was detected by HPTLC
on silica gel 60 F aluminum plates using the mobile phase
hexane-diethyl ether-acetic acid (4:6:0.2) for the unsaponifi-
able portion and (6.5:3.5:0.2) for the saponifiable portion.
Spraying with 10% methanolic sulfuric acid reagent provided
zone detection. Ghee adulteration with tallow down to 20%
was clearly visible in the chromatographic profile.[3]
Dietary supplement analysis
Diazapam was identified as an adulterant in dietary supple-
ments using a combination of HPLC-DAD (diode array
detector), HPLC-MS, and TLC. Fourteen tablet and capsule
supplements were extracted by sonication with methanol,
extracts were separated on silica gel 60 plates by develop-
ment was with hexane-ethyl acetate (1:1), and spots were
detected under 254 nm UV light.[4]
A TLC screening analysis for six synthetic sulfonylurea
type oral antidiabetic agents (tolbutamide, acetohexamide,
chlorpropamide, gliclazide, glibenclamide, and glimepiride)
as adulterants in health food was devised using acetone
shake extraction; silica gel 60 F TLC plates developed with
n-butyl acetate-formic acid (99.6:0.4) for separation of sam-
ples and standards; and detection under 254 nm UV light
and by spraying of three detection reagents: Dragendorff,
10% phosphomolybdic acid (PMA) in methanol, and 30%
sulfuric acid methanol solution. Quantitative analysis was
based on gradient elution HPLC-DAD with a
C18 column.[5]
A multistep analytical method comprising TLC, HPLC-
DAD, electrospray ionization (ESI)-positive ion MS, and one
and two dimensional 1H and 13C nuclear magnetic reson-
ance (NMR) spectrometry was developed for the reliable
determination of the slimming agent sibutramine in
botanical food supplements.[6] The analyte was prepared for
TLC by ultrasonic extraction with chloroform or methanol
JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES 3
and purification by C18 SPE (Chromabond cartridge,
Macherey-Nagel). Samples and standard were chromato-
graphed on silica gel 60 F plates using toluene-methanol
(95:5) mobile phase, and identity was confirmed by compar-
ing Rf values of separated zones detected with Dragendorff
(orange spot) and acidified potassium iodoplatinate (blue
spot) reagents.
Analysis of polyphenols (flavonoids and phenolic acids)
was described as a means of authentication and geographical
traceability of propolis, used in beverages, health foods, and
nutritional supplements, the food industry, and wine.[7] The
proposed HPTLC method for propolis involved silica gel 60
F plates for fingerprint and quantitative analysis. The
method for wine, defined here as a functional food, sug-
gested the use of silica gel sheets with benzene-ethyl acetate-
formic acid (6:3:1) mobile phase, which was chosen as opti-
mal by application of information theory and clustering
methods, identification of substances on the basis of reten-
tion values and UV spectra, and densitometry with scanning
at 250–260 or 350–365 nm.
Fingerprinting of flavonoids from Ginkgo biloba supple-
ments by HPTLC was reported for establishing quality and
authenticity. Capsules, tablets, and liquids were extracted by
sonication with methanol, and extracts were applied as
bands onto Sorbent Technologies nano silica gel F plates.
Development was with dichloromethane-acetonitrile-ethyl
formate-acetic acid-formic acid (11:3.5:3:1.5:1.5), and plates
were imaged by videodensitometry before (254 nm) and after
(366 nm) dipping into natural product (NP) reagent (2-ami-
noethyl diphenylborinate). Ultra-HPLC (UHPLC)-MS ana-
lysis confirmed adulteration of many low quality ginkgo
products with pure flavonoids or extracts made from
another flavonoid-rich material in place of natural flavonol
glycosides in standardized products.[8]
A TLC method was developed for rapid and inexpensive
detection of adulteration of grape seed extract (GSE) by pea-
nut skin extract.[9] GSE, containing proanthocyanidin mono-
mers and oligomers (PACs) as the major bioactive
compounds, was prepared by sonication of capsules with
70% methanol (aqueous) plus 1% added acetic acid. Sample
solutions were spotted manually onto Sorbent Technologies
silica XG F plates and separated by development with acet-
one-acetic acid-toluene (3:1:3). Chromatograms were visual-
ized by spraying with or submerging in vanillin stain (10 g
vanillin/50 mL HCl) and heating. Nine tested products were
found to be adulterated with peanut skin oil based on the
content of PACs at levels not comparable to authentic GSE.
An HPTLC method was devised to permit effective
screening for the presence of three phosphodiesterase type 5
inhibitors (PDE5-Is; sildenafil, vardenafil, and tadalafil) and
eight of their analogs as adulterants in finished products,
including chocolate, instant coffee, syrup, and chewing
gum.[10] Sample preparation was by ultrasonic extraction in
methanol, Rf values of individual bands afforded preliminary
identification of PDE5-Is, scanning densitometry enabled
comparison of unknown compound UV spectra with those
of known standard compounds, and single quadrupole ESI-
MS of zones eluted with a TLC-MS interface confirmed
4 J. SHERMA AND F. RABEL
Figure 1. Scan peaks of three bands obtained from an investigated sample and their UV spectra are shown. The strategy for identification of these unknown
PDE%-Is was to attempt to match the Rf values and UV spectra of available reference standards, followed by TLC-MS for additional information. (Figure reproduced
from Reference 10 with permission of AOAC International).
identity (Figure 1). The method was successfully applied to
45 commercial lifestyle products, 31 of which were shown to
be adulterated with at least one illegal compound (sildenafil,
tadalafil, propoxyphenyl hydroxyhomosildenafil, or dime-
thylsildenfil). Silica gel 60 F Premium Purity glass plates
were developed at 47% humidity with tert-butyl methyl
ether-methanol-ammonia (20:2:1) mobile phase, and chro-
matograms were imaged under 254 and 366 nm UV light.
HPTLC, image processing, and principal component ana-
lysis (PCA) allowed phenolic fingerprinting for determin-
ation of botanical and geographical origin and authenticity
of European propolis.[11] Samples were extracted by sonic-
ation with dichloromethane, and extracts and phenolic
standards were applied as bands onto silica gel plates that
were developed with toluene-ethyl acetate-formic acid
(6:5:1). Dried plates were dipped into NP and polyethylene
glycol (PEG) reagents, images of fluorescent zones were cap-
tured with a Digistore 2 and processed with ImageJ soft-
ware, and PCA was carried out by PLS ToolBox.
Forty seven commercial granulated powders and extracts
of St John’s wort from Germany and America were analyzed
by HPTLC and 1H-NMR spectrometry coupled with multi-
variate analysis software. Samples were prepared by sonic-
ation with methanol and centrifugation and analyzed
according to USP (United States Pharmacopeia) general
chapter 203 on HPTLC of articles of botanical origin.
Samples were applied as bands to silica gel 60 F plates, ethyl
acetate-dichloromethane-water-formic acid-acetic acid
(100:25:11:10:10) was the mobile phase, and visualization by
viewing in white light and 254 nm and 366 nm UV light and
application of NP and PEG dip detection reagents. Both
regulated and unlicensed product consistency was found to
vary significantly, mainly due to adulteration by incorrect
Hypericum species and food dyes.[12]
Adulteration of health care food with insomnia medicines
(triazolam, estazolam, clonazepam, and oxazepam) was
detected using TLC-surface enhanced Raman spectrometry
(SERS). The added drugs were separated on silica gel with
the mobile phase cyclohexane-ethyl acetate-ethanol (5:3.5:2)
and visualized under 254 nm UV light. The SERS excitation
wavelength was 790 nm, and the surface enhancer was aque-
ous phase silver sol. LOD of the illegally added drugs
was 1–4 ng.[13]
Licorice root is a widely used dietary supplement for
treatment of digestive problems, menopausal symptoms, and
cough, and its extract is used to prepare licorice candy and
is added to many herbal tea mixtures for better flavor. Both
the differentiation of licorice roots of Glycyrrhiza glabra and

G. uralensis based on their HPTLC fingerprints viewed
under 254 nm UV light and then under 366 nm UV light
and white light after derivatization with 10% sulfuric acid in
methanol as well as quantitative densitometric determination
at 254 nm to confirm the minimum 2.5% of 18b-glycyrrhizic
acid required by the USP were performed on one silica gel
60 F plate. Sample preparation was based on sonication of
powdered root with 70% ethanol, and applied bands of
extracts and standards were developed with dichlorome-
thane-methanol-water-formic acid (125:75:15:1)
mobile phase.[14]
Edible oil analysis
Fused-silica capillary column GC-FID and selective ion
monitoring (SIM)-MS determination of the triglyceride (TG;
also termed triacylglycerol, TAG) content of olive oil was
shown to provide authentication of virgin oil compared to
oil adulterated with oils such as corn, cottonseed, or linseed.
The total TG fraction was separated from other fractions by
dilution of oil with isopropanol-hexane (1:4), TLC on silica
gel 60 plates with petroleum ether (40–60  C)-diethyl ether-
1% acetic acid (70:30:1) mobile phase, and visualization with
iodine vapors. The TG fraction that migrated the same dis-
tance as a pure TG standard was scraped off and recovered
from the silica gel particles in chloroform to prepare the
solution for GC analysis. The normalized percentage area of
the TG species was sufficient for the method limits used in
this study.[15]
Edible mustard oil adulteration with cheap, nonedible
argemone oil was evaluated by quantification of dihydrosan-
guinarine after conversion to sanguinarine using Soxhlet
extraction with hexane, HPTLC of applied bands on silica
gel 60 plates using hexane-acetone-methanol (16:3:1) mobile
phase, and fluorescence densitometry at 366 nm. Scraped
and eluted bands were further analyzed by MS. The method
can be used routinely to assess adulteration and in forensic
laboratory poisoning cases due to argemone oil toxicity.[16]
Quantitative Ag-TLC of eight samples of TAGs in sun-
flower oil with different linoleic acid content was done on
silica gel G 0.2 mm thick layers impregnated by dipping into
a 0.5% methanolic solution of silver nitrate and continuous
ascending development in an open cylindrical chamber with
petroleum ether-acetone (25:1) and petroleum ether-acet-
one-ethyl acetate (100:5:2 and 50:3:2) mobile phases.
Charring detection of TAGS was by consecutive treatment
of the plate with bromine and sulfonyl chloride vapors
(30 min each) and heating, and quantitative determination
by absorbance densitometry measurement at 450 nm
(Shimadzu 930 densitometer, zig-zag reflection mode). A
characteristic fingerprint of sunflower TAGs was obtained,
and it was not affected by linoleic acid content. It was sug-
gested to be useful in authenticity and adulteration control
of sunflower and olive oils.[17] The same research group
used similar analytical Ag-TLC methods and PTLC of TAGs
on silica gel with hexane-acetone (25:4) mobile phase in a
later study to detect adulteration of olive oil with vege-
table oil.[18]
JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES 5
It was shown that detection of adulteration of virgin olive
oil by sunflower or soybean oil and of corn oil by canola oil
was possible based on the composition of desmethylsterols
determined by isothermal GC-FID with a Zerbon ZB5 capil-
lary column.[19] The unsponifiable matter was fractionated
by PTLC on silica gel plates impregnated with potassium
hydroxide and developed with diethyl ether (65:35). The
desmethylsterol zone detected by spraying with a solution of
0.2% dichlorofluorescein in ethanol was scratched off,
extracted with hot chloroform, and analyzed by GC.
Flavorings analysis
Vanillin and its homolog ethyl vanillin in food flavorings
were determined by TLC on Sorbfil PTSKh-P-V silica gel
plates with a polymer support using the mobile phases hex-
ane-ethyl acetate (8:2 and 9:1) and detection reagent hepta-
none-ethanol-sulfuric acid (4:5:1). After development, plates
were imaged using a Mustek Scan Express 6000P flatbed
scanner, and quantitative analysis was based on measure-
ment of color intensities of sample and standard zones using
Adobe Photoshop 5.0. The method was used in monitoring
the composition of vanilla flavorings, as well as for the
authentication of artificial or identical-to-natural
flavorings.[20]
Detection of banned coumarin in vanilla extract by TLC
was performed on silica gel 60 by co-spotting coumarin
standard as a control, unknown sample, and coumarin
spiked sample with capillary tubes, developing with hexane-
ethyl acetate (8:2), and viewing under UV light. Rf values of
the yellow-green fluorescent zones in the three lanes were
compared, and identification was confirmed by a fluoromet-
ric spot test and low field NMR spectrometry.[21]
Grain analysis
TLC-digital image analysis methods were described for
determination of ergosterol and chitin content of food
grains as an index of fungal contamination. Fungus infected
maize, rice, sorghum, wheat, groundnut, and sunflower food
grains were subjected to a single step liquid/liquid extraction
followed by manual Hamilton microsyringe application of
samples and standards onto Sigma-Aldrich polyester backed
250 mm silica gel G layers. The mobile phase for ergosterol
determination was toluene-acetone (9:1), and density of
fluorescent spots (peak volumes) were recorded using a
CCD (charge couple device)-based digital image analyzer.
Similar methods were used for TLC-digital image analysis
based fluorometric identification and quantification of chito-
san-calcofluor complex.[22]
Honey analysis
A validated HPTLC method was described for detecting
adulteration and QC of Romanian honeys based on fruc-
tose/glucose ratio and amount of sucrose. Samples were pre-
pared by homogenization with methanol, bands were
applied to silica gel aluminum plates that were developed
6 J. SHERMA AND F. RABEL
twice with ethyl acetate-pyridine-water-acetic acid (6:3:1:0.5)
followed by zone visualization with diphenylamine-aniline
reagent, documentation of the plates, and processing of
chromatogram images by videodensitometry.[23]
HPTLC fingerprinting of non-sugar constituents defined
differences in botanical origin for authentication of 10 test
honeys from the United Kingdom (UK), Queensland, and
Australia based on band profiles (Rf values, colors, and
intensities. Honeys were extracted with dichloromethane,
extracts were applied as bands to silica gel 60 F plates that
were developed in an automated chamber with toluene-ethyl
acetate-formic acid (6:5:1), chromatograms were docu-
mented using an imaging device under white light and 254
and 366 nm UV light, and images were digitally processed
using specialized software. Zones were then derivatized with
vanillin spray reagent and documented under white light
and 366 nm UV light.[24]
Milk analysis
An analytical silver ion TLC (Ag-TLC-densitometry) proced-
ure for rapid assessment of the authenticity of questionable
milkfat samples preliminarily converted to fatty acid esters
(FAMEs) was proposed. On the basis of quantitative analysis
of the fatty acid classes, grouped according to the number
and geometry of the double bonds, the levels of trans-mono-
enoic (Mt) and cis,cis-dienoic (Dcc) fatty acids were used as
preliminary markers of authenticity. Then, each of the satu-
rated (S), cis-monoenoic (Mc), and Dcc fractions of butter
samples isolated by preparative Ag-TLC were subjected to
GC analysis of the component fatty acids for confirmation
of authenticity. Ag-TLC-densitometry was done on silica gel
G layers (0.2 mm thickness) impregnated by dipping into
0.5–1% methanoilc silver nitrate solution, continuous devel-
opment with petroleum ether-acetone (100:1.3) mobile phase
in an open cylindrical tank, charring by consecutive treat-
ment with bromine and sulfuryl chloride, and recording and
quantitative measurement of chromatograms by zigzag
reflection scanning at 450 nm (Shimadzu CS-939 densitom-
eter). PTLC was carried out on Ag-coated 1 mm silica gel
preparative layers.[25]
An HPTLC method was developed and validated for rou-
tine monitoring of sucrose, lactose, urea, and machine oil
and applied to analysis of 20 milk samples collected in Agra,
India. Bands of standards and milk without extraction were
applied to silica gel 60 F plates that were developed with
mobile phase in an unsaturated TTC and then scanned in
the visible mode for quantification. The mobile phase and
detection reagent were n-butanol-acetic acid water (2:1:1)
and aniline-diphenylamine-phosphoric acid for sucrose and
lactose, n-propanol-water (8:2) and Ehrlich reagent for urea,
and hexane-diethyl ether-methanol (7:2:1.5) and 5% sulfuric
acid solution for machine oil. Urea and machine oil were
found as adulterants in tested samples from all dairies
except one.[26]
Vegetable oil adulteration of ghee (clarified milk fat) was
determined by RP TLC of unsaponifiable matter from ghee
samples on C18 Fs plates (s designates the presence of acid
stable indicator) developed with petroleum ether-aceto-
nitrile-methanol (2:4:4). Sterols were detected using PMA
reagent. Amounts of adulteration with cheaper oils were
estimated by visual comparison of separated standard and
sample zones to be up to 7.5% coconut oil; up to 1% sun-
flower, soybean, and groundnut oils; and up to 2%
“designer oil”.[27]
Routine analysis of melamine added to milk in order to
increase the apparent protein content for economic adulter-
ation was carried out by HPTLC-densitometry-MS. Market-
purchased milk was filtered and applied as bands with
standards to silica gel 60 F plates that were subsequently
developed with isopropanol-dichloromethane-water (5:2.5:3)
and scanned at 200 nm. Melamine identity was confirmed
by matching sample and standard in situ UV spectra and
elution of zones using of a TLC-MS Interface into a single
quadrupole mass spectrometer and monitoring the charac-
teristic 127.23 amu molecular ion peak (M þ 1).[28]
Cow and buffalo milk fats adulterated with 1% or more
vegetable oils (groundnut, soya bean, and sunflower) were
analyzed on RP18 plates based on the presence of b-sitos-
terol as a marker and some additional spots ascribable to
their occurrence in vegetable oils only. Unsaponifiable mat-
ter from fat samples was extracted with chloroform, extracts
were spotted manually for development with petroleum
ether-acetonitrile-methanol-ethyl acetate (1:1:2:1) or petrol-
eum ether-chloroform-acetonitrile-2-propanol-ethyl acetate
(1.5:0.5:5:3:1), and spots were detected by spraying with a
15% ethanol solution of PMA.[29]
Soft drink and juice analysis
UV-visible spectrometry and MS were used after TLC isola-
tion on silica gel 60 F G plates to prove that orange juices
contain provitamin A carotenoid a-cryptoxanthin accompa-
nying b-cryptoxanthin or the less nutritional nonprovitamin
zeinoxanthin. b-Cryptoxanthin standard was isolated from a
saponified extract of peppers on silica gel 60 G F plates
using petroleum ether (40–60  C)-acetone-diethylamine
(10:4:1) mobile phase, and lutein standard from saponified
extract of spinach leaves and the monohydroxycarotenoid
fraction of orange juice from a saponified extract of ultrafro-
zen juice using diethyl ether mobile phase.[30] It was shown
in a similar study that one of the carotenoids traditionally
thought to occur in orange juices, isolutein, was mis-identi-
fied and was really cis-antheraxanthin. Evidence for this was
obtained by isolation of the diepoxycarotenoid fraction of
orange juice separated by PTLC on 0.5 mm slica gel G F
layers developed with petroleum ether (40–60  C)-acetone
(7:3) by scraping and elution, preparation of antheraxanthin
standard by reaction of zeaxanthin with 3-chloroperoxyben-
zoic acid and purification by PTLC on 0.7 mm layers of sil-
ica gel 60 F developed with petroleum ether (65–95  C)-
acetone-diethylamine (10:4:1), and identification in orange
juice by co-chromatography on silica gel analytical alumi-
num plates developed with diethyl ether mobile phase and
by HPLC-DAD with a C30 column.[31]

HPTLC and headspace (HS)-solid phase microextraction
(SPME)-GC-MS methods to differentiate between authentic
and adulterated (watered down or containing illegal artificial
preservatives) Aloe vera products were reported.[32] Applied
sample bands were resolved on silica gel 60 F plates devel-
oped using n-butanol-n-propanol-acetic acid-water (3:1:1:1).
Chromatograms were detected with a solution of anisalde-
hyde reagent (anisaldehyde-sulfuric acid) by dipping fol-
lowed by heating and comparted to Aloe-typical bands of an
authentic Aloe vera standard to evaluate authenticity.
TLC-densitometry was used to establish the authenticity
of commercial juices and conformity with the label inscrip-
tion based on fingerprints of anthocyanins from ethanolic
berry fruit extracts and anthocyanidins from their hydro-
lyzed products. Macherey Nagel CEL 300-25 cellulose glass
plates were developed with hydrochloric acid-acetic acid-
water (10:1:3) in a saturated glass chamber. Quantitative
determination was performed by densitometry in the reflect-
ance-absorption mode at 520 nm. A complementary HPLC
method with an ODS-2 Hypersil column, gradient elution
with different ratios of acetonitrile and phosphate buffer,
and detection at 520 nm was also described.[33]
TLC on silica gel 60 F plates detected the marker 30 50 -di-
C-b-glucopyranosylphloretin (PD) present in calamondin
juice adulterating shiikuwasha juice for economic gain.[34]
The mobile phase was chloroform-methanol-1% phosphoric
acid (65:35:5), and visualization of separated PD was by
spraying 10% sulfuric acid and heating.
Eleven anthocyane pigments in juice, pomace, animal
feed, and wine were separated on silica gel 60 F plates with
ethyl acetate-2-butanone-formic acid-water (7:3:1.2:0.8)
mobile phase for anthocyanins and ethyl acetate-toluene-for-
mic acid-water (10:3:0.8:1.2) for anthocyanidins in a cham-
ber with humidity adjusted to 33% by the presence of
saturated magnesium chloride hydrate. Detection was by
means of DPPH (2,2-diphenyl-1-picrylhydrazyl radical)
reagent and Vibrio fischeri reagent in a CAMAG
BioLuminizer. Zones were identified by direct elution into
an Agilent single quadrupole-ESI mass spectrometer. The
fully validated effect directed analysis (EDA) method
allowed discrimination between plant sources of products
for authentication and characterization of unknown ingre-
dients in the foods.[35]
Tartrazine, amaranth, sunset yellow, and brilliant blue
synthetic food dyes were determined in commercial carbo-
nated orange and grape soft drinks by TLC after SPE on
C18 Sep-Pak cartridges. Extracts and standards were applied
with a capillary onto silica gel plastic backed plates, and col-
orants were separated with isopropanol-ammonia (8:3)
mobile phase in a glass chamber and identified by compari-
son of zone Rf values. Quantitative determination was per-
formed by HPLC-DAD. The methods were applied to
confirm that the dye composition found matched that stated
on product labels and complied with the legislative
requirements.[36]
Direct manual spotting with a micropipet, silica gel 60 F
plates, toluene-ethyl acetate-formic acid (8:4:1) mobile phase
in a glass tank, and viewing under 254 nm UV light and
JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES 7
then under 366 nm light after spraying with NP reagent
were used to separate and identify the phenolic compounds
ellagic acid and gallic acid in pomegranate juice. Their anti-
oxidant activity was evaluated with DPPH spray reagent.
These two phenolics can serve as markers in authenti-
city studies.[37]
TLC confirmed that permitted food dyes claimed on the
labels (e.g., sunset yellow and tartrazine) were found in
most branded soft drinks and juices but illegal colors that
could harm health were in many local drinks collected in
Pakistan. AOAC 2005 standard food dye extraction method
comprising addition of 30 mL of 5% acetic acid solution to
50 mL of sample, adding wool thread to soak 15 min while
boiling, and eluting the trapped dyes with 1 M ammonia
was used before TLC with silica gel stationary phase, pro-
panol-ammonia (60:15) mobile phase, and identification by
comparing Rf values of standard and separated sam-
ple dyes.[38]
A method was developed and validated for b-carotene
determination from dietary supplements and juices contain-
ing carrots, bananas, and apples.[39] Dietary supplement cap-
sules and tablets were extracted by shaking with ethyl
acetate and the juices with chloroform. Of the mobile phases
tested, the optimum separation was obtained on aluminum
oxide 60 F neutral (type E) layers activated with methanol
and the mobile phase chloroform-methanol-acetone-ammo-
nia (10:22:53:0.2). Densitometry was performed in the
absorbance mode at 450 nm. TLC fingerprints of all extracts
were obtained to give reliable authentication of identity.
Spice analysis
SFE with carbon dioxide was used to detect the adulteration
of black pepper powder with ground papaya seed. TLC ana-
lysis of the SFE extracts on silica gel 60 plates with ethylene
dichloride mobile phase showed a fluorescent band under
366 nm UV light at Rf 0.17 that proved to be a marker for
the presence of papaya with an LOD of 20 g/kg. GC-MS
identified the straight chain aldehydes n-nonanal, n-decanal,
and n-dodecanal as components in the marker zone.[40]
Turmeric powders from markets in India were analyzed
for curcumin and metanil yellow as a measure of quality by
ethanol extraction, development of silica gel 60 F aluminum
sheets with chloroform-methanol (9:1) mobile phase, and
densitometry at 420 nm. A significant number of non-
branded powders were found to contain curcumin amounts
below the major brand minimum of 2.1% and 1–8.5 mg/g of
the extraneous dye, which may cause a health threat.[41] A
“2-directional” HPTLC method reported earlier by the same
laborator.[42] for the simultaneous qualitative and quantita-
tive determination of curcumin, metanil yellow, and sudan I
and IV in mixed curry powders is rather complex and
poorly described, and it is not a true “two-dimensional
(2D)” metho.[43] in which one sample is applied in the cor-
ner of a plate and developed with two different mobile
phases with complementary selectivities at right angles to
increase resolution of mixtures, and the standards and sam-
ples are not applied together and developed identically in
8 J. SHERMA AND F. RABEL
parallel as is required for reliable quantitative HPTLC-
densitometry.
HPTLC determination of protocatechuic acid present in
methanol and acetone Soxhlet extracts of the spice and con-
diment Amomum subulatum Roxb. (Greater Cardamom) at
levels of 1.05 and 0.863% was the basis of its authentication.
The validated method included silica gel 60 F 0.2 mm thick-
ness plates, chloroform-acetic acid (9:1) mobile phase in a
TTC at 40% humidity, and scanning at 254 nm for
quantification.[44]
TLC fingerprints containing the pungent compounds 6-,
8-, and 10-gingerol; 10-shogaol; and five other discrete
zone.[45] were characteristic for the authentication of
Jamaican ginger powders were extracted by shaking with
methanol according to the ISO 13685:1997(E) method,
bands were applied to silica gel plates that were developed
with hexane-ethyl acetate-formic acid (55:40:5), and chroma-
tograms were visualized with ammonium molybdate-sulfuric
acid and p-anisaldehyde-sulfuric acid-acetic acid spray
reagents. The gingerols and shogaol can be quantified by
HPTLC to confirm authenticity by extraction of the samples
four times by sonication and vortexing with methanol,
application of samples and standards as round spots on
LiChrospherV Si 60 F aluminum plates (spherical particle
R
silica gel layer) using a CAMAG Nanomat, separation with
toluene-ethyl acetate (3:1) mobile phase, and absorbance
reflection densitometry at 577 nm after spraying with anisal-
dehyde-sulfuric acid reagent.[46]
In a study of saffron authentication from different ori-
gins.[47] dried and ground stigmas were extracted with etha-
nol-water (8:2) by vortexing and ultrasonication, and the
extracts separated by TLC on silica gel G F plates using
development in a horizontal chamber with n-butanol-acetic
acid–water (4:1:1). Chromatograms were photographed with
a Canon digital camera model Isus 115 Hs, and intensity
profiles of RGB (red, green, blue) characteristics were pro-
duced and processed by specially designed image analysis
software based on MATLAB Version 7. The method allowed
comparison of different types of saffron from Iran and
determination of various adulteration colorants.
Sudan I is a carcinogenic and mutagenic azo dye used as
a common adulterant in spices to impart a desirable red
color. MIP (molecularly imprinted polymer)-TLC was
employed to determine Sudan I adulteration in paprika
powder down to a level of 1 ppm (or 2 ng/spot) by using
Sudan I imprinted polymer as the stationary phase to select-
ively retain dye spotted at the origin with little interference
after development using hexane-chloroform (1:1).
Synthesized gold colloid was applied onto the original spot
to enhance SERS intensity for Sudan I. PCA plot and partial
least squares regression models were constructed and a lin-
ear regression model correlated spiking levels of 5–100 ppm
with peak intensity of Sudan I spectra. Separation (30–40 s)
and detection (1 or 0.1 s) were extremely fast by using both
commercial bench top and custom made portable Raman
spectrometers.[48]
TLC of 80% methanol extracts of ground spice samples
spotted on silica gel 60 plates, developed with cyclohexane-
dichloromethane-ethyl acetate-methanol (5:1:4:0.8), and
visualized under 366 nm UV light and after spraying with
vanillin-sulfuric acid produced fingerprints with species spe-
cific spots of alkaloids, flavonoids, saponins, steroid glyco-
sides, and tannins. These were found helpful for
standardization and authentication of the spice and condi-
ment Allspice grown in Sri Lanka.[49]
TLC-image analysis with chemometric techniques was
applied to validating the authenticity of saffron and identify-
ing the adulterants (e.g., sumac, turmeric, safflower, com-
mon madder, quinolone yellow, sunset yellow, and
tartrazine). Extraction by sonication with ethanol-water
(8:2); silica gel 60 F plates and n-butanol-acetic acid-water
(4:1:1) mobile phase; image capture with an Apple iPhone
65 camera; processing of images by MATLAB environment
software (R2016b); and different unsupervised and super-
vised chemometric methods, variable selection, and linear
discriminant analysis (LDA) were applied in this QC
method for Iranian sourced saffron in the international
food market.[50]
Caffeine impregnated HPTLC silica gel plates (Macherey-
Nagel Nano-SIL-PAH) were used in a new method to deter-
mine the eight most frequently found unauthorized azo
dyes, i.e., para red and sudan I, II, III, IV, 7B, orange G,
and red B, in spices, spice mixtures, pastes, sauces, and
palm oils. A simple post-chromatographic UV irradiation
provided cleanup of sample extracts, which took 4 min for
up to 46 samples in parallel. Twenty three simultaneous sep-
arations with isohexane-ethyl methyl ketone (5:1) in an
automated chamber controlled at 45% humidity could be
carried out within 20 min for quantification or 46 separa-
tions within 5 min for screening. Linear (4–40 ng/band) and
polynomial (10–200 ng/band) calibration curves at
390–550 nm had high densitometric correlation coefficients
and low RSDs; LOD and LOQ were 2–3 and 6–9 ng/zone,
respectively; and recoveries of dyes from samples of
70–120% at spike levels of 25–50 mg/kg and 150–300 mg/kg.
Identification of the dyes was by elution with a TLC-MS
Interface through a short C18 HPLC column to remove coe-
luted caffeine from the layer into a single quadrupole ESI
mass spectrometer.[51]
Sweetener analysis
Carbohydrate profiles obtained by TLC and PCA were
found helpful for the authentication, characterization, and
subsequent detection of intentional adulteration of maple
syrup and distinguishing it from other natural sweeteners
(agave, sugarcane, corn, and honey). Additional information
about the type, amount, and difference in content of carbo-
hydrates in the syrups was obtained by anion exchange col-
umn chromatography coupled to pulsed amperometric
detection (HPAEC-PAD) and Fourier transform infrared
(FTIR) spectrometry. Syrup solution samples were applied
to a silica gel aluminum plate, developed with butanol-pro-
panol-water, and sprayed with the reagent aniline-diphenyl-
amine-phosphoric acid in acetone for carbohydrate
visualization.[52]

Tea analysis
An HPTLC fingerprinting method was established for
authentication of 12 commercially available fruit teas (rasp-
berry, strawberry, blueberry, rose hip, and wildberry). The
fingerprints of red fruit and tea extracts obtained in visible
light and fluorescence at 366 nm UV light after derivatiza-
tion with NP and PEG reagents indicated different charac-
teristic colored zones allowing a clear differentiation
between all teas in terms of their type and producer using
PCA of chromatogram image data, obtained using ImageJ
computer software, performed using the STATISTICA 7
package. Fruit and teas were extracted by maceration with
acidified ethanol (0.1% HCl), and extract bands were applied
to silica gel 60 F aluminum plates that were developed with
n-butanol-formic acid-water (12:3:4.5).[53]
A validated HPTLC method was reported for quantifica-
tion of 7-O-glucoside (active marker) and fingerprinting of
commercial chamomile tea products. The method was com-
pared to an HPLC method given in the European
Pharmacopoeia and found superior for adulteration evalu-
ation. Silica gel 60 NH2 F HPTLC plates combined with pat-
tern recognition techniques found that not tea bags but
crude flowers sold on the market were adulterated with
other plant material. Wild chamomile, chamomile like sam-
ples, and tea products were ultrasonically extracted by
methanol, and extracts were membrane filtered. Samples
were applied as bands onto plates that were developed with
ethyl acetate-formic acid-acetic acid-water (30:1.5:1.5:3)
mobile phase. NP and PEG dip reagents were used for
detection, plates were documented at 366 nm, and quantifi-
cation was performed via a quadratic calibration curve rang-
ing from 10 to 50 ng/band.[54]
Vegetable analysis
Oxytocin is used as an adulterant in vegetable and fruit
samples to increase growth rate and in milk to enhance pro-
duction from lactating animals. A validated HPTLC method
was devised to determine oxytocin and applied to market-
purchased brinjal (spicy eggplant) and bottle gourd vegeta-
bles, watermelon and papaya fruit, and milk. The method
comprised vegetable and fruit extraction with methanol by
percolation, sonication, and soaking, and dilution of milk
with methanol; application of samples and standards band-
wise onto silica gel 60 F plates; methanol-ammonia (9:1)
adjusted to pH 6.8 with sodium bicarbonate-sodium hydrox-
ide buffer as the mobile phase; and densitometry at the
absorption peak maximum of 270 nm. Oxytocin was found
in watermelon at a 75.2 mg/kg level and milk at 39.1 mg/kg.
Papaya and brinjal were not adulterated.[55]
Thickening agent analysis
HPTLC was applied in combination with pattern recogni-
tion techniques for authentication of highly similar thicken-
ing agents, which are mainly based on polysaccharides or
biopolymers. After methanolysis, the monomer units of the
JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES 9
thickeners were separated on HPTLC silica gel 60 plates
using i-propyl acetate-ethyl acetate-methanol-water
(5:4:1:0.1) mobile phase and detected by derivatization with
aniline diphenylamine o-phosphoric acid reagent.
Documentation was performed under white light. According
to their resulting fingerprint and chemical pattern, the
agents were classified in several groups by PCA and hierar-
chic cluster analysis.[56]
Wine and other alcoholic beverage analysis
Biogenic amines and resveratrol were quantified in 25 differ-
ent Hungarian wines from the same region using OPLC.
PCA was able to characterize the wines according to con-
tents of biogenic amines and polyphenols (determined by
the Folin-Ciocalteu spectrometric method) and distinguish
red and white wines. Derivatization of biogenic amines with
dansyl chloride was carried out before analysis using a BS
50 personal OPLC chromatograph with HPTLC silica gel 60
F layers and two step gradient elution with hexane-n-buta-
nol-triethylamine (90:10:8.1) followed by n-hexane-n-butanol
(8:2). Densitometric evaluation of chromatograms was at
300 nm in the fluorescence mode. Resveratrol was analyzed
by linear isocratic development with chloroform-methanol
(100:8) mobile phase and densitometry at 305 nm.[57]
Purification by SPE on C18 Supelco cartridges and then
HPTLC on C18 W F plates using methanol—water-tri-
fluoroacetic acid (55:45:1) enabled clear separation and
accurate quantification of the three classes of anthocyanins
contained in Vitis vinifera grapes and in red wines produced
from them. The ester group on the glucose unit was found
to be the key factor enabling separation of glycosides, acetyl-
glycosides, and coumaroylglycosides. Differentiation of dif-
ferent vintages and an overview of the pigment profile
during aging were determined. Twenty samples were ana-
lyzed within a few hours.[58]
TLC was used for separation and identification of phen-
olic compounds in samples of two red wines commercially
available in Croatia.[59] Sample extraction was by liquid-
liquid extraction with diethyl ether at pH 2. Samples applied
at bands were developed on silica gel 60 F plates with ben-
zene-ethyl acetate-formic acid (6:3:1), the most efficient of
11 mobile phases evaluated by information theory and clus-
tering methods. Phenolic compounds were detected in chro-
matograms by their visible colors or by treatment with
ammonia vapor followed by inspection under 254 and
366 nm UV light before and after spraying with 1% etha-
nolic aluminum chloride. Six compounds were identified:
gallic acid, caffeic acid, apigenin, kaempferol, p-coumaric
acid, and naringenin. Results serve as baseline information
for studies of wine authenticity.
The sedative and hypnotic agent chloral hydrate is added
to alcoholic beverages to increase their potency and
make the drink addictive. A TLC method was described for
detection of chloral hydrate in a toddy with 5% alcoholic
content involving filtration of the drink, application of a
sample and standards manually with a micropipette on
silica gel G glass plates, development in a chamber with
10 J. SHERMA AND F. RABEL
hexane-acetone-methanol (8:3:0.5), and spraying with
orcinol under alkaline conditions to give a yellow fluores-
cent spot of chloral hydrate with an LOD of 10 mg. Other
adulterants of alcoholic beverages, e.g., saccharin, diazepam,
and phenobarbitone, do not interfere.[60]
The origin of Romanian red wines (Cabernet Sauvignon,
Merlot, and Burgundy) and their adulteration were moni-
tored based on separation of pigments.[61] Samples were dir-
ectly applied as bands with a microsyringe to aluminum
backed silica gel RP18 F plates that were developed in an N-
chamber with acetonitrile-water-formic acid (20:29:1) mobile
phase. Separated zones were detected in visible light and
under 366 nm UV light. The pigments were mostly phenols
with antioxidant activity that was evaluated by use of
DPPH spray reagent.
A TLC method based on the European Pharmacopeia
was adapted and a simple and economical HPTLC assay was
developed in order to determine absinthin in alcohol bever-
ages. Also, the bitter principle of wormwood was used for
the first time to assess the authenticity of absinthe. Samples
prepared by extraction of absinthe with dichloromethane
were applied bandwise onto silica gel 60 F glass plates that
were developed with acetone-acetic acid-toluene-dichlorome-
thane (1:1:3:5). Bands were detected by dipping into acetic
anhydride-sulfuric acid-ethanol (1:1:10) reagent and heating,
and densitometry was used for quantification of brown
zones of absinthin at 554 nm and recording spectra of bands
in the 400–700 nm range for identification. Using external
standards of wormwood extracts, the concentration of
wormwood in the absinthe samples was determined.[62]
Dansyl derivatives of biogenic amines were separated on
silica gel 60 glass plates developed with chloroform-triethyl-
amine (4:1). The detection was done at 312 nm, and images
captured by a Vilber-Lourmat Infinity-1000 system were
analyzed by means of BIO-1D software. Histamine, tyram-
ine, putrescine, and cadaverine were determined at concen-
trations between 1 and 20 mg/L. This method is high
throughput, inexpensive, and simple for identifying undesir-
able wines containing levels of amines high enough to cause
health problems.[63]
Levels of gallic acid, caffeic acid, resveratrol, and rutin
phenolic antioxidants linked to the health benefits of
wine were quantified in 43 red and two white wines by
application without pretreatment along with standards as
bands on silica gel 60 F plates developed in a two-step gra-
dient comprising dichloromethane-methanol-formic acid
(7.5:2:0.75) followed by a water in oil microemulsion con-
sisting of sodium dodecyl sulfate-pentanol-water-heptane
(8 g:25 mL:8 mL:160 mL). Images of plates were captured
under 366 nm UV light before and after spraying with NP
reagent, and collection and transformation of images into
chromatograms and quantitative evaluation were made using
image analysis software. Concentrations of the analytes were
higher in red compared with white wines and varied with
type and country of origin.[64] The Agatonovic-Kustrin
research group later used the same TLC techniques and che-
mometrics (PCA and artificial neural network: ANN) to
characterize the wines as a function of grape variety and for
their classification, discrimination, and authentication.[65]
An HPTLC fingerprint method was developed for
authentication of three varieties of Romanian white wines,
produced in three different years in different vineyards.
After application of wines as bands, silica gel 60 F layers
were developed with ethyl acetate-formic acid-acetic acid-
water (10:1:1:2) and RP18 layers with methanol-water-acetic
acid (5:5:0.1). Images of chromatograms taken under 254
and 366 nm UV light after detection with NP and PEG dip
reagents were digitally processed using ImageJ software. The
digitized data and cluster analysis were used to distinguish
wines by their variety, vineyards, and vintage and to authen-
ticate closely related white wines.[66] The same research
group performed fingerprinting plus antioxidant activity
evaluation of 27 Romanian red wines with the same techni-
ques on the silica gel 60 F plates by adding detection with
DPPH dip reagent. The fingerprints were useful for QC of
wines and their differentiation as well for finding that anti-
oxidant activity increased in the order Feteasca Neagra,
Carernet Sauvignon, and Merlot.[67]
Conclusions and future prospects
It is documented in the references cited above and the
others following that TLC/HPTLC can be successfully
applied for the determination of many different classes of
compounds in order to authenticate and determine adulter-
ation of all types of foods,[68–71] beverages.[72] and dietary
supplements.[68] requiring minimal sample preparation and
providing high sample throughput and low cost
per sample.
It is certain that food authentication will be significantly
aided by greater use of TLC coupled online with MS, usually
through a commercial TLC-MS interface for direct elution
from the layer into the mass spectrometer inlet. As an
example, different levels of adulteration of olive oils were
recognized by laser desorption ionization (LDI) MS directly
from a sample spot on a silica gel 60 TLC plate.[73] This
method could perhaps have provided even more analytical
information if the plate was developed with a mobile phase
before spectra were recorded from the separated zones.
Chemometric pattern recognition methods combined
with TLC e.g..[74] is a very powerful tool that will be applied
in the future more often for characterization and authentica-
tion of foods. As examples of chemometric applications in
addition to those cited above, the TLC profiles of intra- and
interday precision for the natural food preservative and food
flavoring Piper bele L. (PBL) folium methanol extract were
studied for their peak marker recognition and identification.
The numerical chromatographic parameters of the peak
markers, hierarchical clustering analysis (HCA) and PCA
were applied to authenticate PBL folium extract from
another Piper species folium extract. The spotted extract was
developed on silica gel 60 F plates with the mobile phase
toluene-ethyl acetate (93:7), images of the plate were cap-
tured, the plate was scanned by densitometry in the absorp-
tion and fluorescent modes, and in situ spectra were

recorded between 190 and 400 nm.[75] Also, efficient detec-
tion of small quantities of hazelnut oil in virgin olive oil was
affirmed by silica gel PTLC of oil samples for isolation of
TAGs, diacylglycerols, and 1,2-phosphatidic acids prior to
high resolution GC-FID, with elaboration of the analytical
data using LDA and ANN.[76]
Image analysis is being more reported each year, although
slit scanning densitometry is still most widely used for
accurate and precise quantification in food analysis. One
reason for this increasing use of image analysis is the greater
availability of digital image analysis software, some of which
is free of charge open source such as ImageJ and
quanTLC.[77] A chapter on TLC image processing and che-
mometrics that is in a new book on chemometrics in chro-
matography, which is included in the Chromatographic
Science Series edited by Nelu Grinberg.[78] should be a cata-
lyst for further applications of both image analysis and che-
mometrics in TLC.
TLC will complement much more sophisticated and
expensive LC-MS/MS method.[79] that have been developed
for detection of one or more food allergens in a single
run.[80] This technology, which is quickly replacing ELISA
(enzyme linked immunosorbent assay) and PCR (polymerase
chain reaction) as the most used methods for food allergen
detection.[81] has the additional benefit of allowing detection
of protein based adulteration (food fraud). Examples are
identification of peanuts in a product labeled to contain
only pine nuts,[82,83] and determination of fish species by
species specific peptides, allowing identification of the pres-
ence or absence of fish species and also the replacement of a
high quality species with a low quality species.[84] AOAC
has recognized a protein-based LC-MS/MS method for mul-
tiple allergen detection as a First Action method (not yet
published), and European government working groups have
been formed to develop and validate methods for food aller-
gen detection and food fraud testing. Additional examples of
food fraud analysis by LC-MS/MS have been published for
bovine, pig, fish, and milk products.[85] plant-based prod-
ucts.[86] and herbs and spices.[87]
Acknowledgments
We thank Karen F. Haduck, Lafayette College Interlibrary Resource
Sharing Specialist, for her invaluable work in obtaining copies of many
of the publications cited in this chapter, without which it could not
have been written.
References
[1] Sherma, J. Thin Layer Chromatography in Food and
Agricultural Analysis. J. Chromatogr. A. 2000, 880, 129–147.
[2] Jurado, J. M.; Jimenez-Lirola, A.; Narvaez-Rivas, M.; Gallardo,
E.; Pablos, F.; Leon-Camacho, M. Characterization and
Quantification of 4-Methylsterols and 4,4-Dimethylsterols from
Iberian Pig Subcutaneous Fat by Gas Chromatography-Mass
Spectrometry and Gas Chromatography-Flame Ionization
Detector and Their Use to Authenticate the Fattening Systems.
Talanta. 2013, 106, 14–19.
[3] De, S.; Nariya, P.; Jirankalgikar, N. Development of a Novel
High Performance Thin Layer Chromatographic-Densitometric
JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES 11
Method for Detection of Tallow Adulteration in Cow Ghee. J.
Planar Chromatogr. Mod. TLC. 2013, 26, 486–490.
[4] Mikami, E.; Goto, T.; Ohno, T.; Oka, H.; Kanamori, H.
Simultaneous Analysis Seven Benzodiazepines in Dietary
Supplements as Adulterants Using High Performance Liquid
Chromatography and Its Application to an Identification
System for Diazepam. J. Health Sci. 2005, 51, 278–283.
[5] Kumasaka, K.; Kojima, T.; Honda, H.; Doi, K. Screening a
Quantitative Analysis for Sulfonyl Type Oral Antidiabetic
Agents in Adulterated Health Food Using Thin Layer
Chromatography and High Performance Liquid
Chromatography. J. Health Sci. 2005, 51, 453–460.
[6] Csupor, D.; Boros, K.; Danko, B.; Veres, K.; Szendrei, K.;
Hohmann, J. Rapid Identification of Sibutramine in Dietary
Supplements Using a Stepwise Approach. Pharmazie. 2013, 68,
15–18.
[7] Medic-Saric, M.; Bojic, M.; Rastija, V.; Cvek, J. Polyphenolic
Profiling of Croatian Propolis and Wine. Food Technol.
Biotechnol. 2013, 51, 159–170.
[8] Avula, B.; Sagi, S.; Gafner, S.; Upton, R.; Wang, Y. -H.; Wang,
M.; Khan, H. A. Identification of Ginkgo biloba Supplements
Adulteration Using High Performance Thin Layer
Chromatography and Ultra High Performance Liquid
Chromatography-Diode Array Detector-Quadrupole Time of
Flight Mass Spectrometry. Anal. Bioanal. Chem. 2015, 407,
7733–7746.
[9] Villani, T. S.; Reichert, W.; Ferruzzi, M. G.; Pasinetti, G. M.;
Simon, J. E. Chemical Investigation of Commercial Grape Seed
Derived Products to Assess Quality and Detect Adulteration.
Food Chem. 2015, 170, 271–280.
[10] Do, T. T. K.; Theocharis, G.; Reich, E. Simultaneous Detection
of Three Phosphodiesterase Type 5 Inhibitors and Eight of
Their Analogs in Lifestyle Products and Screening for
Adulterants by High Performance Thin Layer Chromatography.
J. AOAC Int. 2015, 98, 1226–1233.
[11] Milojkovic-Opsenica, D.; Ristivojevic, P.; Trifkovic, J.; Vovk, I.;
Lusic, D.; Tesic, Z. TLC Fingerprinting and Pattern Recognition
Methods in the Assessment of Authenticity of Poplar Type
Propolis. J. Chromatogr. Sci. 2016, 54, 1077–1083.
[12] Booker, A.; Agapouda, A.; Frommenwiler, D. A.; Scotti, F.;
Reich, E.; Heinrich, M. St John’s Wort (Hypericum perforatum)
Products – an Assessment of Their Authenticity and Quality.
Phytomedicine. 2018, 40, 158–164.
[13] Li, J. H.; Cheng, N. N.; Liu, J. C.; Li, L.; Jia, S. S. Rapid On-Site
TLC-SERS Detection of Four Sleep Problems Drugs Used as
Adulterants in Health Care Food. Spectrosc. Spect. Anal. 2018,
38, 1122–1128.
[14] Widmer, V. M.; Frommenwiler, D. A. Qualitative and
Quantitative HPTLC Analysis of Licorice Root. CAMAG
Bibliography Service CBS. 2018, 120, 12–13.
[15] Andrikopoulos, N. K.; Giannakis, L. G.; Tzamtzis, V. Analysis
of Olive Oil and Seed Oil Triglycerides by Capillary Gas
Chromatography as a Tool for the Detection of Adulteration of
Olive Oil. J. Chromatogr. Sci. 2001, 39, 137–145.
[16] Ghosh, P.; Reddy, M.M.K; Sashidhar, R. B. Quantitative
Evaluation of Sanguinarine as an Index of Argemone Oil
Adulteration in Edible Mustard Oil by High Performance Thin
Layer Chromatography. Food Chem. 2005, 91, 757–764.
[17] Marekov, I.; Tarandjiiska, R.; Momchilova, S.; Nikolova-
Damyanova, B. Quantitative Silver Ion Thin Layer
Chromatography of Triacylglycerols from Sunflower Oils
Differing in the Level of Linoleic Acid. J. Liq. Chromatogr.
Relat. Technol. 2008, 31, 1959–1968.
[18] Marekov, I.; Panayotova, S.; Tarandjiiska, R. Silver Ion TLC of
Minor Triacylglycerol Components for Unambiguous Detection
of Adulteration of Olive Oil with Vegetable Oil. J. Liq.
Chromatogr. Relat. Technol. 2010, 33, 1013–1027.
[19] Youseff, R.; Soubh, L.; Alassaf, Z. Detection of Vegetable Oil
Adulteration Using Desmethylsterols Composition. Int. J.
Pharm. Sci. Rev. Res. 2014, 28, 229–233.
12 J. SHERMA AND F. RABEL
[20] Gerasimov, A. V.; Gornova, N. V.; Rudometova, N. V.
Determination of Vanillin and Ethylvanillin in Vanilla
Flavorings by Planar (Thin Layer) Chromatography. J. Anal.
Chem. 2003, 58, 758–765.
[21] Garcia, O.; Keeney, C.; Coticone, S. Detection of Coumarin in
Artificially Adulterated Vanilla Bean Extracts in a Forensic
Science Laboratory. Chem. Educator. 2015, 20, 227–228.
[22] Kosuri, T.; Nayak, S.; Gul, M. Z.; Rupula, K.; Beedu, S. R. TLC-
Digital Image-Based Fluorometric Analysis of Ergosterol and
Chitin Content in Food Grains Artificially Infested with
Aspergillus flavus and Fusarium verticilloides. Food Anal. Meth.
2018, 11, 1267–1280.
[23] Puscas, A.; Hosu, A.; Cimpoiu, C. Application of a Newly
Developed and Validated High Performance Thin Layer
Chromatographic Method to Control Honey Adulteration. J.
Chromatogr. A. 2013, 1272, 132–135.
[24] Locher, C.; Neumann, J.; Sostaric, T. Authentication of Honeys
of Different Floral Origins via High performance Thin Layer
Chromatographic Fingerprinting. J. Planar Chromatogr.-Mod.
TLC. 2017, 30, 57–62.
[25] Marekov, I.; Nedelcheva, D.; Panayotova, S.; Tarandjiiska, R.
Detection of Milkfat Adulteration by GC Analysis of Saturated,
cis-Monoenoic and cis,cis-Dienoic Fatty Acid Fractions Isolated
By Silver Ion TLC. J. Liq. Chromatogr. Relat. Technol. 2011, 34,
888–901.
[26] Rani, R.; Medhe, S.; Raj, K. R.; Srivastava, M. M. High
Performance Thin Layer Chromatography for Routine
Monitoring of Adulterants in Milk. Natl. Acad. Sci. Lett. 2012,
35, 309–313.
[27] Rani, A.; Sharma, V.; Arora, S.; Lal, D.; Kumar, A. A Rapid
Reversed Phase Thin Layer Chromatographic Protocol for
Detection of Adulteration in Ghee (Clarified Milk Fat) with
Vegetable Oils. J. Food Sci. Technol. 2015, 52, 2434–2439.
[28] Rani, R.; Medhe, S.; Srivastava, M. HPTLC-MS Analysis of
Melamine in Milk: Standardization and Validation. Dairy Sci. &
Technol. 2015, 95, 257–263.
[29] Upadhyay, N.; Kumar, A.; Rathod, G.; Goyal, A.; Lal, D.
Development of a Method Employing Reversed Phase Thin
Layer Chromatography for Establishing Milk Fat Purity with
Respect to Adulteration with Vegetable Oils. Int. J. Dairy
Technol. 2015, 68, 207–217.
[30] Melendez-Martinez, A. J.; Britton, G.; Vicario, I. M.; Heredia, F.
J. Identification of Zeinoxanthin in Orange Juices. J. Agric.
Food Chem. 2005, 53, 6362–6367.
[31] Melendez-Martinez, A. J.; Britton, G.; Vicario, I. M.; Heredia, F.
J Identification of Isolutein (Lutein Epoxide) as cis-
Antheraxanthin in Orange Juice. J. Agric Food Chem., 2005, 53,
9369–9373.
[32] Lachenmeier K.; Kuepper, U.; Musshoff, F.; Madea, B.; Reusch,
H.; Lachenmeier, D. W. Quality Control of Aloe Vera
Beverages. EJEAFChe 2005, 4, 1033–1042.
[33] Filip. M.; Vlassa, M.; Copaciu, F.; Coman, V. Identification of
Anthocyanins and Anthocyanidins from Berry Fruits by
Chromatographic and Spectroscopic Techniques to Establish
Juice Authenticity from Market. J. Planar Chromatogr.-Mod.
TLC. 2012, 25, 534–541.
[34] Yamamoto, K.; Yahada, A.; Sasaki, K.; Ogawa, K.; Koga, N.;
Ohta, H. Chemical Markers of Shiikuwasha Juice Adulterated
with Calamondin Juice. J. Agric. Food Chem. 2012, 60,
11182–11187.
[35] Krueger, S.; Urmann, O.; Morlock, G. E. Development of a
Planar Chromatographic Method for Quantitation of
Anthocyanes in pomace, feed, juice and wine. J. Chromatogr. A.
2013, 1289, 105–118.
[36] de Andrade, F. I.; Guedes, M. I. F.; Viera. I. G. P.; Mendes, F.
N. P.; Rodrigues, P. A. S.; Maia, C. S. C.; Avila, M. M. M.; de
Matos Ribeiro, L. Determination of Synthetic Food Dyes in
Commercial Soft Drinks by TLC and ion-Pair HPLC. Food
Chem. 2014, 157, 193–198.
[37] Fakhri, E.; Petroczi, A.; Naughton, D. P. Assessing the Efficacies
of Phenolic Compounds in Pomegranate Juice Using Thin
Layer Chromatography. Acta. Chromatogr. 2014, 26, 563–573.
[38] Naseem, Z.; Alim-un-Nisa, Z. F.; Kalim, I.; Saeed, K.
Identification of Synthetic Food Dyes in Beverages by Thin
Layer Chromatography. Pak. J. Food Sci. 2015, 25, 178–181.
[39] Starek, M.; Guja, A.; Dabrowska, M. Assay of b-Carotene in
Dietary Supplements and Fruit Juices by TLC-Densitometry.
Food Anal. Methods. 2015, 8, 1347–1355.
[40] Bhattacharjee, P.; Singhal, R. S.; Gholap, A. S. Supercritical
Carbon Dioxide Extraction for Identification of Adulteration of
Black Pepper with Papaya Seeds. J. Sci. Food Agric. 2003, 83,
783–786.
[41] Dixit, S.; Purshottam, S. K.; Khanna, S. K.; Das, M. Surveillance
of the Quality of Turmeric Powders from City Markets of India
on the Basis of Curcumin Content and the Presence of
Extraneous Colours. Food Addit. Contam. 2009, 26, 1227–1231.
[42] Dixit, S.; Khanna, S. K.; Das, M. A Simple 2-Directional High
Performance Thin Layer Chromatographic Method for the
Simultaneous Determination of Curcumin, Metanil Yellow, and
Sudan Dyes in Tumeric, Chili, and Curry Powders. J. AOAC
Int. 2008, 91, 1387–1396.
[43] Rabel, F.; Sherma, J. A Review of Advances in Two
Dimensional Thin Layer Chromatography. J. Liq. Chromatogr.
Relat. Technol. 2016, 39, 627–639.
[44] Shivanand, P.; Mahalaxmi, R. Identification and Determination
of Protocatechuic Acid Present in Greater Cardamom Fruit
Extracts by HPTLC Technique. Int. J. Pharm. Sci. Rev. Res.
2010, 1, 27–32.
[45] Salmon, C. N. A.; Bailey-Shaw, Y. A.; Hibbert, S.; Green, C.;
Smith, A. M.; Williams, L. A. D. Characterisation of Cultivars
of Jamaican ginger (Zingiber officinale Roscoe)by HPTLC and
HPLC. Food Chem. 2012, 131, 1517–1522.
[46] Melianita, F.; Witha, J.; Arifin, S.; Kartinasari, W. F.;
Indrayanto, G. Simultaneous Densitometric Determination of 6-
Gingerol, 8-Gingertol, 10-Gingerol, and 6-Shogaol in Some
Commercial Gingers. J. Liq. Chromatogr. Relat. Technol. 2009,
32, 567–577.
[47] Djozan, D.; Karimian, G.; Jouyban, A.; Iranmanesh, F.;
Gorbanpour, H.; Alizadeh-Nabil, A. Discrimination of Saffron
Based on Thin Layer Chromatography and Image Analysis. J.
Planar Chromatogr.-Mod. TLC. 2014, 27, 274–280.
[48] Gao, F.; Hu, Y.; Chen, D.; Li-Chan, E.C.Y.; Grant, E.; Lu, X.
Determination of Sudan I in Paprika Powder by Molecularly
Imprinted Polymer-Thin Layer Chromatography-Surface
Enhanced Raman Spectroscopic Biosensor. Talanta. 2015, 143,
344–352.
[49] De Soysa, E.J.S.; Abeysinghe, D. C.; Dharmadasa, R. M.
Comparison of Phytochemicals Antioxidant Activity and
Essential Oil Content of Pimenta dioica (L.) Merr. (Myrtaceae)
with Four Selected Spice Crop Species. World J. Agric. Res.
2016, 4, 158–161.
[50] Sereshti, H.; Poursorkh, Z.; Aliakbarzadeh, G.; Zarre, S. Quality
Control of Saffron and Evaluation of Potential Adulteration by
Means of Thin Layer Chromatography-Image Analysis and
Chemometrics Methods. Food Control. 2018, 90, 48–57.
[51] Schwack, W.; Pellissier, B.; Morlock, G. Analysis of
Unauthorized Sudan Dyes in Food by High Performance Thin
Layer Chromatography. Anal. Bioanal. Chem. https//doi.org/10.
1007/s00216-018-0945-6.
[52] Mellado-Mojica, E.; Seeram, N. P.; Lopez, M. G. Comparative
Analysis of Maple Syrups and Natural Sweeteners:
Carbohydrates Composition and Classification (Differentiation)
by HPAEC-PAD and FTIR Spectroscopy-Chemometrics. J.
Food Compos. Anal. 2016, 52, 1–8.
[53] David, L.; Hosu, A.; Moldovan, B.; Cimpoiu, C. Evaluation and
Authentication of Red Fruits Teas by High Performance Thin
Layer Chromatography Fingerprinting. J. Liq. Chromatogr. Relat
Technol. 2014, 37, 1644–1653.

[54] Guzelmeric, E.; Ristivojevic, P.; Vovk, I.; Milojkovic-Opsenica,
D.; Yesilada, E. Quality Assessment of Marketed Chamomile
Tea Products by a Validated HPTLC Method Combined with
Multivariate Analysis. J. Pharm. Biomed. Anal. 2017, 132,
35–45.
[55] Rani, R.; Medhe, S.; Raj, K. R.; Srivastava, M. Standardization
of HPTLC Method for the Estimation of Oxytocin in Edibles. J.
Food Sci. Technol. 2013, 50, 1222–1227.
[56] Ristivojevic, P.; Morlock, G. E. High Performance Thin Layer
Chromatography Combined with Pattern Recognition
Techniques as Tool to Distinguish Thickening Agents. Food
Hydrocoll. 2017, 64, 78–84.
[57] Csomos, E.; Heberger, K.; Simon-Sarkadi, L. Principal
Component Analysis of Biogenic Amines and Polyphenols in
Hungarian Wines. J. Agric. Food Chem. 2002, 50, 3768–3774.
[58] Lambri, M.; Jourdes, M.; Glories, Y.; Saucier, C. High
Performance Thin Layer Chromatography (HPTLC) Analysis of
Red Wine Pigments. J. Planar Chromatogr.-Mod. TLC. 2003,
16, 88–94.
[59] Rastija, V.; Mornar, A.; Jasprica, I.; Srecnik, G.; Medic-Saric, M.
Analysis of Phenolic Components in Croatian Red Wines by
Thin Layer Chromatography. J. Planar Chromatogr.-Mod. TLC.
2004, 17, 26–31.
[60] Mali, B. D.; Garad, M. V. Thin Layer Chromatographic
Detection of Chloral Hydrate in an Alcoholic Beverage. J.
Planar Chromatogr.-Mod. TLC. 2005, 18, 397–399.
[61] Cimpoiu, C.; Hosu, A.; Briciu, R.; Miclaus, V. Monitoring the
Origin of Wine by Reversed Phase Thin Layer
Chromatography. J. Planar Chromatogr.-Mod. TLC. 2007, 20,
407–410.
[62] Lachenmeier, D. W. Assessing the Authenticity of Absinthe
Using Sensory Evaluation and HPTLC Analysis of the Bitter
Principle Absinthin. Food Res. Int. 2007, 40, 167–175.
[63] Romano, A.; Klebanowski, H.; La Guerche, S.; Beneduce, L.;
Spano, G.; Murat, M.-L.; Lucas, P. Determination of Biogenic
Amines in Wine by Thin Layer Chromatography/Densitometry.
Food Chem. 2012, 135, 1392–1396.
[64] Agatonovic-Kustrin, S.; Hettiarachchi, C. G.; Morton, D. W.;
Razic, S. Analysis of Phenolics in Wine by High Performance
Thin Layer Chromatography with Gradient Elution and High
Resolution Plate Imaging. J. Pharm. Biomed. Anal. 2015, 102,
93–99.
[65] Agatonovic-Kustrin, S.; Milojkovic-Opsenica, D.; Morton, D.
W.; Ristivojevic, P. Chemometric Characterization of Wines
According to Their HPTLC Fingerprints. Eur. Food Res.
Technol. 2017, 243, 659–667.
[66] Hosu, A.; Cimpoiu, C. HPTLC Fingerprinting: A Useful Tool
for White Wines Authentication. J. Liq. Chromatogr. Relat.
Technol. 2016, 39, 303–307.
[67] Hosu, A.; Danciu, V.; Cimpoiu, C. Validated HPTLC
Fingerprinting and Antioxidant Activity Evaluation of Twenty-
Seven Romanian Red Wines. J. Food Compos. Anal. 2015, 41,
174–180.
[68] Mankar, S. N.; Banerjee, D.; Vaidya, P. S.; Husain, M. I.;
Jaiswal, P. K. Aflatoxin B1 in Amla Whole (Dry), Amla
Powder, Caster Seed Shikakai, Mahua and Tamarind. J. Inst.
Chemists (India). 2005, 77, 169–170.
[69] Teo, P.; Ma, F.; Liu, D. Evaluation of Taurine by HPTLC
Reveals the Mask of Adulterated Edible Bird’s Nest. J. Chem.
2013, Article ID 325372, http://dx.doi.org/10.1155/2013/325372.
[70] Upadhyay, N.; Kumar, A.; Goyal, A.; Lal, D. A Planar
Chromatographic Method to Detect Adulteration of Vegetable
Oils in Ghee. J. Planar Chromatogr.-Mod. TLC. 2014, 27,
431–437.
[71] Peng, G.; Chang, M.; Fang, M.; Liao, C.; Tsai, C.; Tseng, S.;
Kao, Y.; Chou, H.; Cheng, H. Incidents of Major Food
JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES 13
Adulteration in Taiwan Between 2011 and 2015. Food Control.
2017, 72, 146–152.
[72] Hosu, A.; Cimpoiu, C. Thin Layer Chromatography Applied in
Quality Assessment of Beverages Derived From Fruits. J. Liq.
Chromatogr. Relat. Technol. 2017, 40, 239–246.
[73] de Oliveira, D. N.; Catharino, R. R. Direct Metabolic
Fingerprinting of Olive Oils Using STELDI-MS. J. Food
Compos. Anal. 2015, 38, 131–134.
[74] Bosque-Sendra, J. M.; Cuadros-Rodriguez L.; Ruiz-Samblas, C.;
de la Mata, A. P. Combining Chromatography and
Chemometrics for the Characterization and Authentication of
Fats and Oils from Triacylglycerol Compositionsl Data-A
Review. Anal. Chim. Acta. 2012 724, 1–11.
[75] Wirasuta, M. A. G., Srinadi, G. A. M.; Dwidasmara, I. B. G.;
Ardiyanti, N. L. P. P.; Trisnadewi, G. A. A.; Paramita, N. L. P.
V. Authentication of Piper betle L. Folium and Quantification
of Their Antifungal Activity. J. Tradit. Complement. Med. 2017,
7, 288–295.
[76] Damiani, P.; Cossignani, L.; Simonetti, M. S.; Del Signore, A.;
Giaccio, M.; Damiani, F.; Favretto, L. G. Stereospecific Analysis
of Triacylglycerol Fraction to Detect Small Amounts of
Hazelnut Oil in Virgin Olive Oil. J. Commodity Sci. 2001, 40,
95–114.
[77] Fichou, D.; Morlock, G. E. quanTLC, an Online Open Source
Solution for Videodensitometric Quantification. J. Chromatogr.
2018, 1560, 78–81.
[78] Hemmateenejad, S.; Talebanpour, B. Rafatmah, E.; Shojaeifard,
Z.; Mobaraki, N.; Yousefinejad, S. Chemometrics and Image
Processing in Thin Layer Chromatography. In Chemometrics in
Chromatography, Komsta, L., Vander Heyden, Y., Sherma, J.,
Eds.; CRC Press/Taylor & Francis Group: Boca Raton, FL,
2018; pp 469–494.
[79] Brockmeyer, J. Novel Approaches for MS Based Detection of
Food Allergens: High Resolution, MS3, and Beyond. J. AOAC
Int. 2018, 101, 124–131.
[80] Boo, C. C.; Parker, C. H.; Jackson, L. S. A Targeted LC-MS/MS
Method for the Simultaneous Detection and Quantitation of
Egg, Milk, and Peanut Allergens in Sugar Cookies. J. AOAC Int.
2018, 101, 108–117.
[81] Diaz-Amigo, C.; Popping, B. A Global Reflection on Food
Allergen Regulations, Management, and Analysis. J. AOAC Int.
2018, 101, 1.
[82] Korte, R.; Lepski, S.; Brockmeyer, J. Comprehensive Peptide
Marker Identification for the Detection of Multiple Nut
Allergens Using a Non-Targeted LC-HRMS Multi-Method.
Anal. Bioanal. Chem. 2016, 408, 3059–3069.
[83] Korte, R.; Brockmeyer, J. MRM3 Based LC-MS Multi-Method
for the Detection and Quantification of Nut Allergens. Anal.
Bioanal. Chem. 2016, 408, 7845–7855.
[84] Korte, R.; Monneuse, J. M.; Gemrot, E.; Metton, I.; Humpf, H.
U.: Brockmeyer, J. New High Performance Liquid
Chromatography Coupled Mass Spectrometry Method for the
Detection of Lobster and Shrimp Allergens in Food Samples via
Multiple Reaction Monitoring and Multiple Reaction
Monitoring Cubed. J. Agric. Food Chem. 2016, 64, 6219–6227.
[85] Marchis, D.; Altomare, A.; Gili, M.; Ostorero, F.; Khadjavi, A.;
Corona, C.; Ru, G.; Cappelletti, B.; Gianelli, S.; Amadeo, F.;
et al. LC-MS/MS Identification of Species-Specific Muscle
Peptides in Processed Animal Proteins. J. Agric. Food Chem.
2017, 65, 10638–10650.
[86] Huschek, G.; Bonick, J.; Merkel, D.; Huschek, D.; Rawel, H.
Authentication of Leguminous Based Products by Targeted
Biomarkers Using High Resolution Time of Flight Mass
Spectrometry. LWT-Food. Sci. Technol. 2018, 90, 164–171.
[87] Galvin-King, P.; Haughey, S. A.; Elliott, C. T. Herb and Spice
Fraud; the Drivers, Challenges and Detection. Food Control.
2018, 88, 85–97.