WELCOME

TO

HPTLC SPECIALISTS
 AIM OF TRAINING

Familiarize the
participants with the state
of the art of High
Performance Thin Layer
Chromatography.
ANCHROM
 ANCHROM VISION

To make MODERN TLC the

first and most preferred
method of analysis in all the
markets we operate in.

ANCHROM
 ANCHROM
¾ DEDICATED TO HPTLC (1985)
¾ MR. DILIP CHAREGAONKAR, TECHNOCRAT FOUNDER
¾ CAMAG ASSOCIATION SINCE 1978
¾ PRODUCT + SERVICES PACKAGE

BUSINESS POLICY
EQUAL EMPHASIS

LAB. SUPPORT

SALES & MARKETING SERVICE

TECHNOLOGISTS, NOT TRADERS
ANCHROM
 ANCHROM 3M
SUPPORT
MAN - UNLIMITED TRAINING
LIBRARY
PRETRAINED STAFF
FREE RECRUITMENT
METHOD – METHOD DEVELOPMENT
METHOD VALIDATION
WORKING STANDARDS
MACHINE – QUALIFICATION
AMC
STOCK – SPARES, CONSUMABLES
COMPONENT LEVEL SERVICE
FIELD UPGRADATION h/w & s/w

ANCHROM
 , SWITZERLAND
¾ DEDICATED TO HPTLC (1960)
¾ DR. DIETER JÄNCHEN, TECHNOCRAT FOUNDER
¾ AT BASEL (HEART OF SWISS CHEMICAL INDUSTRY)
¾ CLOSE CO- ORDINATION WITH CIBA, SANDOZ,
ROCHE, MERCK, BAYER, BASF ETC.)
¾ CBS JOURNAL – SINCE 1965
¾ PIONEERED MOST NEW PRODUCTS
¾ APPLICATION LABS – SWITZERLAND, USA & INDIA
¾ 12 % TURNOVER SPENT IN R & D
¾ WITH ANCHROM SINCE 1978

ANCHROM
PROPRIETORY TECHNOLOGIES
¾ HPTLC SYSTEM MANAGER.
¾ FULLY AUTOMATIC TLC SCANNER.
¾ VARIO CHAMBER FOR METHOD DEVELOPMENT &
SCREENING.
¾ AMD SYSTEM FOR GRADIENT HPTLC
¾ LIBRARY OF SPECTRA OF FORENSIC INTEREST,
WITH SEARCH .
¾ AUTOMATIC PLATE COATER.
¾ TRANSILLUMINATOR ( 302 NM) COMBINED WITH
TLC / HPTLC PHOTODOC.
¾ MICRO – PREPARATIVE CHROM. WITH SAME
HPTLC SYSTEM.
¾ PHOTO DOCUMENTATION WITH INDUSTRIAL CAMERA.

ANCHROM
 SUMMARY OF ANALYTICAL
CHEMISTRY
ORIGIN OF SAMPLES

BIOLOGICAL INDUSTRIAL
INDUSTRIAL LABS

Q.C. R&D PROCESS ENVIRO.

WET CHEM. INSTRUMENTATION BIOLOGICAL

PHYSICAL

SPECTROSCOPY CHROMATOGRAPHY OTHERS

UV HPLC < AAS >
IR HPTLC X - RAY
NMR GLC THERMAL ETC.
< MS > IC +
GPC “ HYPHENATED”
ANCHROM
ANCHROM
 HPTLC
COMPLIMENTARY
TO GLC / HPLC

NO CHROM. LAB
IS
COMPLETE
W/O HPTLC ANCHROM
 CHROMATOGRAPHY
AIM

MIXTURE

SEPARATE

IDENTIFY QUANTIFY MICRO

- PREP

COMPARE ABSOLUTE
( UV – VIS ) ( IR / MS ) ANCHROM
QUANTITATIVE TLC

HPTLC TLC – FID / FPD

CHROMATOGRAPH

ANCHROM
HPTLC

ANCHROM
 H P T L C HISTORY
1951 KIRCHNER
1957 STAHL “ HANDBOOK OF TLC ”
1975 PRECOATED PLATES, HPTLC
1980 INSTRUMENTATION, CAMAG
1985 GRADIENT HPTLC
1990 AUTOMATION
1996 VIDEO DOCUMENTATION
2000 SYSTEM MANAGER
2005 12-bit DIGITAL PHOTODOC.
2005 AUTO DEVELOPING CHAMBER

ANCHROM
PRACTICE OF HPTLC
( SIMPLIFIED SAMPLE PREPARATION )

METHOD CREATION

SAMPLE APPLICATION

CHROMATOGRAPHY DEVELOPMENT

VISUALISATION ( DOC / DERIV )

SCANNING
( QUANTIFICATION & IDENTIFICATION )

PHOTO DOC
POST CHROM DERIV

SCANNING PHOTO DOC

ANCHROM
 HPTLC
ADVANTAGES
“WHY FIRST CHOICE”
¾ FASTEST
¾ OFF – LINE
¾ VISIBLE
¾ SIMPLEST
¾ LOW RUNNING COST
¾ NO RISK ANALYSIS
¾ DOUBLE CONFIRMATION

ANCHROM
 Stationary Phase
Chemically well defined mostly porous inorganic
substances with high surface area are used.
The performance of TLC sorbent depends on
surface area of the particle, pore volume, mean pore
diameter, uniform particle size.
Small particle size increases separation efficiency,
resolution & decreases analysis time.
Polarity decreases as shown :
Silica > amino silica > cyano silica > octadecyl silica
Polar Med. Polar Non-Polar

ANCHROM
 Stationary Phase
¾ Silica Gel (fairly Universal)
¾ 60ºA, 7 µm
¾ Zinc silicate with Mn
¾ 0.2 mm layer

ANCHROM
 Vapour Phase
¾ The vapor phase in the developing chamber.
The gas phase can heavily affect the
chromatographic result.

¾ Adjustment of the chromatographic properties

of the stationary phase in TLC/HPTLC through
exposure to a defined gas phase (relative
humidity, solvent composition, etc.).

ANCHROM
 Mobile Phase
¾ The mobile phase is the solvent or solvent mixture moving
through the stationary phase on the TLC/HPTLC plate during
development.
¾ Mobile phase should be chosen taking into consideration
chemical properties of analytes & sorbent layers.
¾ Use of mobile phase containing more than three or four
components should be avoided as it is often difficult to get
reproducible ratios of different components.
¾ Advantages:
Mobile phase evaporates before derivatization
Does not interfere with determination of the position
of solute spots/ bands
Smaller volume of mobile phase required.

ANCHROM
 Solvent Polarity
To increase Rf increase polarity & vice versa
¾ n- Hexane 0.1 ¾ Ethanol 4.3
¾ Cyclohexane 0.2 ¾ Ethyl acetate 4.4
¾ Carbon disulphide 0.3 ¾ Ethyl methyl ketone 4.7
¾ Carbon tetrachloride 1.6 ¾ Dioxane 4.8
¾ Isopropyl ether 2.4 ¾ Acetone 5.1
¾ Toluene 2.4 ¾ Methanol 5.1
¾ Chlorobenzene 2.7 ¾ Pyridine 5.3
¾ Benzene 2.7 ¾ Acetonitrile 5.8
¾ Diethyl ether 2.8 ¾ Acetic acid 6.0
¾ Dichloromethane 3.1 ¾ Nitromethane 6.0
¾ 1,2- dichloroethane 3.5 ¾ Aniline 6.3
¾ 2- propanol 3.9 ¾ Ethylene glycol 6.9
¾ Tetrahydrofuran 4.0 ¾ Dimethyl sulphoxide 7.2
¾ Chloroform 4.1 ¾ Water 10.2

ANCHROM
 Reference to Front (Rf)

The position of
any fraction in Solvent front

TLC is Fraction

characterized by
a b
Rf. App. Position

Rf = a / b
Rf value can be
from 0 to 1.
ANCHROM
 HPTLC COMPLETE
SYSTEM

¾ SYSTEM MANAGER - DOCUMENTATION

¾ SAMPLE BAND APPLICATOR
¾ CHROM. DEVELOPMENT – CHAMBERS
¾ VISUALISATION – UV CABINET
¾ QUANT. EVALUATION – SCANNER
¾ PHOTODOCUMENTATION

ANCHROM
ANCHROM
 SAMPLE APPLICATION CHROM. DEVELOPMENT
DEVICES DEVICES

HPTLC
SYSTEM
POST
SCANNER MANAGER CHROM.
DEVICES
S/W

winCATS

CONTROL AND OR DOCUM.

ANCHROM
 STEP 1

CREATE METHOD
winCATS

ANCHROM
 STEP 2

SAMPLE
APPLICATION

ANCHROM
HPTLC SAMPLE APPLICATION
SPOT
NON – UNIFORM
VS
UNIFORM
BAND
9
DISTRIBUTION

DIFFUSSION COUNTER – DIFF.

¾ ENLARGED ¾ COMPACT
¾ BETTER RESOLVED

LOW VOLUME LARGER VOLUME

¾ NO OVERLOAD
¾ BETTER SENSE
¾ BETTER PRECISION

SCAN PROBLEMS NO PROBLEM

¾ NOT UNIFORM SENSE ¾ UNIFORM SENSE
¾ BEAM / SPOT ¾ ALIQUOT SCAN
ALIGNMENT
ANCHROM
 HPTLC QUANTIFICATION
SPOT VS BAND
9
BROAD SHARP
PEAK PEAK

BAND –
HIGHER Rf WIDTH
BIGGER ALMOST
DIAMETER SAME

HIGHER
SENS
BEAM/SPOT
ALIGN. ALIQUOT
SCAN

ANCHROM
 LINOMAT 5
SAMPLE APPLICATION TECHNIQUE
QUANTITATIVE ANALYSIS
Most often used. Usual application
volume 6 – 20 ul. Usual band size 6 mm.
For aq. samples 8 mm, distance from side.
12 – 15 mm, distance from bottom – 8 mm.
MICRO – PREP. ISOLATION
For isolation of fractions on a mg scale (for
identification by spectroscopy). Usual band length
– 190 mm. Usual volume applied 500 ul
( = 5 – 20 mg ).
IN – SITU CLEAN – UP
Used to separate ( extracted ) fatty matrix
from the sample. Samples applied 110 mm
from bottom, developed in ether, plate dried,
cut 120 mm from bottom, turned 180˚ and
developed with appropriate mobile phase.
SUPERIMPOSE
Required for overlapping of internal std. or
spiking or derivatizing reagent or improving
quantification; Single method for
application and superimpose.
ANCHROM
AUTOMATIC TLC SAMPLER 4

ANCHROM
 LINOMAT 5

4 – MODE APPLICATOR
ANCHROM
SAMPLE APPLICATION
SUMMARY
¾ BAND APPLICATION, NOT SPOTS

¾ ACCURATE POSITIONING

¾ ACCURATE VOLUMES

¾ SHARP START ZONES

ANCHROM
 STEP 3

CHROMATOGRAM

DEVELOPMENT

ANCHROM
CHROMATOGRAM
DEVELOPMENT
ALL PHASES ACTIVE

¾STATIONARY
¾MOBILE

AND
¾VAPOUR

ANCHROM
 HPTLC
DEVELOPMENT CHAMBERS
¾ TWIN TROUGH CHAMBERS
¾ VARIO METHOD DEVELOPMENT
¾ HORIZONTAL DEVELOPMENT
¾ AUTOMATIC MULTIPLE
DEVELOPMENT ( GRADIENT )
¾ VERTICAL / HORIZONTAL
¾ SATURATION / SANDWICH

ANCHROM
 TWIN TROUGH CHAMBERS

(4-15 ml)

CIBA PATENT
ANCHROM
VARIO CHAMBER

VARIO
CHROMATOGRAM
METHOD DEVELOPMENT
ANCHROM
 AUTOMATIC
DEVELOPING CHAMBER
¾Automated
¾Reproducible
¾Humidity control
¾GLP
¾System manager control

ANCHROM
GRADIENT CHAMBER

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 GRADIENT H P T L C
WHERE TO USE :
¾ HIGH RESOLUTION –
POLYMERS, ENVIRONMENT, N.P.
¾ SCREENING –
AZO DYES, N.P., BIOTECH
¾ CONSTANT Rf / COMPLEX MATRIX –
FORENSIC, DOPING
¾ TRACE ANALYSIS –
PESTICIDE ETC RESIDUES
¾ MULTIPOLAR SAMPLES –
LIPIDS, NATURAL PRODUCTS
¾ CLOSELY RELATED COMPOUNDS –
ISOMERS, ORG. SYNTHESIS
¾ UNIVERSAL GRADIENT – PILOT RUN ANCHROM
 GRADIENT
¾ Up to 25 developments
¾ In same direction
¾ Polar Non- polar
¾ Each time front moves further
¾ Computer controlled
¾ All polarity covered
¾ Up to 40 components separated

ANCHROM
Stage 1
Normal universal gradient
based on dicloromethane, gas
phase basic
Stage 2
Universal gradient based on
diisopropylether
Stage 3
From these conclusions the gradient
depicted below was defined. Three
isocratic dev. steps with methanol
were found to be necessary to ensure
that no substance remains stationary
at the line of sample application
where it could entrap analytes

SUBSTANCE 1 AND 2 DIFFER IN ONLY 1 METHYL GROUP. ANCHROM

 HPTLC
CHROMATOGRAM
DEVELOPMENT
SUMMARY
¾ All 3 PHASES ACTIVE
¾ CAPILLARY FORCE ONLY
¾ ANY COMBO : SP / MP/ VP
¾ ISOCRATIC OR GRADIENT
¾ HORIZONTAL / VERTICAL
ANCHROM
STEP 4
POST
CHROMATOGRAPHY
a. VISUALISATION

b. PHOTO DOCUMENTATION

c. DERIVATISATION

ANCHROM
VISUALISATION
UV CABINET

254 + 366 nm. SAFE

ANCHROM
UV 366nm Longwave

Observe first at 366nm. Acclimatize

eyes for 30 sec. before judgement.
Check each sample for fluorescence!
ANCHROM
UV 254nm Shortwave

Bright background (green or blue).

Fluoroscence indicator F254. Fractions
appear dark
ANCHROM
 PHOTODOCUMENTATION
WITH INDUSTRIAL CAMERA

ANCHROM
PHOTODOCUMENTATION

¾ CONVINCING DATA
¾ VISUAL CONFIRMATION
¾ 254 + 366 nm, UV, VISIBLE
¾ GLP

ANCHROM
POST – CHROMATOGRAM
DERIVATIZATION
FOR

¾ SPECIFICITY
¾ ULTRA HIGH SENSITIVITY
¾ ADDITIONAL INFORMATION
¾ VISUAL CONFIRMATION
(Rarely required if scanner available)

ANCHROM
 DERIVATIZATION
TECHNIQUES
SPRAY
NOT UNIFORM
DIP
UNIFORM
9
UNCONTROLLED CONTROLLED

POLLUTION NO POLLUTION

MORE PRECISE

ANCHROM
DERIVATISATION

IMMERSION DEVICE ANCHROM

 POST
CHROMATOGRAPHY
SUMMARY
¾ UV CABINET - VISUALISATION

¾ PHOTODOCUMENTATION

¾ DERIVATIZATION

ANCHROM
 STEP 5

QUANTITATIVE
EVALUATION
ANCHROM
 HPTLC
QUANTITATIVE EVALUATION

SCAN
+
SPECTRUM

SCANNER 3
OPTIONS
MWL
SPECTRUM LIBRARY
TRACK OPTIMISATION
DUAL WAVELENGTH
TRANSMISSION

ANCHROM
SYSTEM MANAGER

32 BIT S/W
ANCHROM
 SCANNER
¾ FULLY AUTOMATIC SCAN + SPECTRA
¾ ULTRA FAST SCAN & SPECTRUM
¾ BEAM / SAMPLE ALIGNMENT CHECK
¾ GLP
¾ UV – VIS – FLUOR. REFLECTANCE
¾ 190 – 800 NM RANGE
¾ SPOT CHECK
¾ LAMP USE TRACKED

ANCHROM
 D2 ENTRANCE LENS SYSTEM

MONOCHROMATOR ENTRY SLIT

W MONOCHROMATOR GRATING
Hg

LAMP LENS SYSTEM

SELECTOR MACRO = TLC
MICRO = HPTLC

MIRROR
MIRROR
DISK WITH
SLIT
APERTURES

REFERENCE BEAM
PHOTO - SPLITTER
MULTIPLIER

MEASURING
SCANNER PHOTOMULTIPLIER

OPTICS SCANNING OBJECT

PHOTODIODE
(TRANSMISSION)
ANCHROM
 Signal to Noise Ratio
(S/N)
Expression :
S/N = Es / (Ss2 + Sbk2 )1/2

Es – Analytical signal obtained by subtracting

the blank signal from the total
Ss – Standard deviation of the analytical
signal
Sbk - Standard deviation of the blank signal

ANCHROM
SCANNED DATA
(Quantification)

ANCHROM
CALIBRATION CURVE

ANCHROM
SPECTRA (IDENTIFY)
e.g. Food colour
std

sample

Spectra of allura red & sample at Rf 0.45

ANCHROM
 SCANNER S/W OPTION
QUALIFICATION
¾ 17 MAIN TESTS & 200 SUB – TESTS; LAMP ALIGNMENT AUTO CORRECT
¾ MECHANICALS / OPTICS / ELECTRONICS TESTED; RECOM. – ONCE A YEAR

200 TESTS

ANCHROM
 SCANNER S/W OPTION
MULTIWAVE SCAN
¾ 31 WAVELENGTHS
¾ λ MAX OF EACH FRACTION
¾ DATA STORED FOR EVALUATION

A
N
C
H
R
O
QUANTIFICATION / FINGERPRINT M
 Dual Wavelength

¾ The subtraction result does not produce any

meaningful data unless the lamp,
measurement mode, optical filter are identical
for both measurements, and the detector mode
is set to automatic.

ANCHROM
 SCANNER S/W OPTION
SPECTRUM LIBRARY
¾ CREAT YOUR OWN LIBRARY
¾ SEARCH - λ MAX + RF
¾ LIBRARY OF SPECTRA ( CAMAG )
¾ LIBRARY OF SPECTRA ( ANCHROM )

IDENTIFICATION ANCHROM
 SCANNER S/W OPTION
TRACK OPTIMISATION
¾ FOR SPOTS. NOT BANDS NON- GLP
¾ ALL FRACTIONS OF ALL SAMPLES OPTIMISED
¾ USE ONLY IF SAMPLE EXHAUSTED

ANCHROM
 DENSITOMETRY
SUMMARY
¾ AUTO SCAN - QUANTIFICATION
¾ AUTO SPECTRA – IDENTIFICATION
¾ UV – VIS - FLUOR
¾ REFLECTANCE
¾ 2 SECS / SAMPLE !

ANCHROM
MICRO – PREPARATIVE
CHROMATOGRAPHY
¾ ISOLATION FOR IDENTIFICATION BY
OTHER TECHNIQUES

¾ 5 – 25 mg sample loading

¾ 0.25 TO 2 mm thick LAYER

¾ MANY FRACTIONS COLLECTED

ANCHROM
MICRO – PREPARATIVE CHROMATOGRAPHY

ANCHROM
 HPTLC
MULTI LEVEL
ANALYTICAL TECHNIQUE

TLC ( CONVENTIONAL )

INSTRUMENTAL TLC

HPTLC

GRADIENT H P T L C
ANCHROM
HPTLC FINGERPRINT PROCEDURE
PREPARED SAMPLE

CHROMATOGRAPH

PHOTODOC – 254, 366 nm

GLP
DERIVATISATION MULTIWAVE + FLOUR SCAN

PHOTODOC SPECTRA – IDENTITY PURITY

SCAN / PHOTODOC
FLUOR VISIBLE

ANCHROM
 HERBAL
CHARACTERISATION

1 FINGERPRINT

2 QUANTITATIVE ANALYSIS

e.g. ANDROGRAPHIS
PANICULATA

ANCHROM
 HERBAL ANALYSIS
FINGERPRINT
e.g. ANDROGRAPHIS P.
METHOD DEVELOPMENT
P. 1 - CHROM. METHOD DEVELOPMENT
P. 2 - MULTIWAVELENGTH SCAN (190–350 nm)
P. 3 - MULTIWAVELENGTH SCAN (190–270 nm)
FINGERPRINT
P. 4 - UV SPECTRUM OF - ALL PEAKS
P. 5 - MULTIWAVELENGTH SCAN
( 202, 229, 255 & 279 nm )
P. 6 - FLUORESCENCE SCAN (366 nm)
P. 7 - DIGITAL PHOTODOC.
( 254nm + 366nm & VISIBLE )

ANCHROM
 CHROMATOGRAPHY
METHOD DEVELOPMENT

ANCHROM
MULTIWAVE SCAN AT 20 nm INTERVAL. WAVELENGTH
230 nm SEEMS TO GIVE MAXIMUM NUMBER OF PEAKS.

ANCHROM
 SPECTRA OF MAJOR PEAKS DETECTED AT 230 nm. THE λ max OF
PEAKS ARE THE WAVELENGTH CHOSEN FOR ACTUAL MULTIWAVELENGTH
ANALYSIS e.g. PEAK AT Rf 0.16 HAS λ max OF 280 nm.
ANCHROM SO 280 nm IS TO BE CHOSEN AS ONE OF THE MWLs.
 MULTIWAVE MEASUREMENT FOR ACTUAL ANALYSIS. BY
CHOOSING THE λ max OF EACH PEAK, IT IS ENSURED THAT ALL
IMPORTANT PEAKS ARE DETECTED AT THEIR BEST WAVELENGTH.
USUALLY BETWEEN 3 TO 5 WAVELENGTHS IS SUFFICIENT.
ANCHROM
FLUORESCENCE SCAN FOR FINGERPRINT.
GIVES UNIQUE INFORMATION.
ANCHROM
PHOTODOCUMENTATION

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 QUANTITATIVE HERBAL
ANALYSIS
e.g. ANDROGRAPHOLIDE FROM
ANDROGRAPHIS P.
P. 1 - SCAN OF ALL TRACKS( 229 nm )
P. 2 - 3 - SPECTRUM MATCH OF
ANDROGRAPHOLIDE
STD. PEAK VS SAMPLE
P. 4 - 8 - ANALYSIS METHOD / REPORT
P. 9 - CALIBRATION CURVE
ANCHROM
 IDENTITY

MATCHING OF SPECTRA OF KNOWN FRACTIONS WITH

THE CORRESPONDING FRACTION IN UNKNOWN TO
ENSURE THAT THEY ARE THE SAME SUBSTANCES.
ANCHROM
 PEAK PURITY

PURITY CHECK OF A FRACTION IS DONE BY RECORDING ITS ABS.

SPECTRA AT THREE DIFFERENT PLACES i.e. PEAK MAX PLUS BOTH
THE FLANKS. COMPUTER COMPARES THE THREE AND GIVES
CORREATION FACTORS. THIS TEST CHECKS FOR HOMOGENITY OF
THE FRACTION. ANCHROM
 MULTILEVEL CALIBRATION USING KNOWN
CONCENTRATIONS OF ANDROGROPHOLIDE, IS
NECESSARY TO QUANTIFY IT FROM THE SAMPLES.
ANCHROM
 C A M A G TLC Evaluation Software
***********************************************************

ANCHROM
ANCHROM
Impurity Profiling

ANCHROM
 Known Impurity Quantification

Sample NB 362 Step 1 impurity Step 2 impurity

Spectra Step 1 impurity Spectra Step 2 impurity Image(254nm)

ANCHROM
 Impurity Quantification by
(Universal) Dilution Method

U1 : Batch No.1 : 800 ug

U2 : Batch No. 2 : 800ug
U3 : Batch No. 3 : 800ug
E : REF. SOLUTION ‘E’ : 0.4ug(0.05%)
D : REF. SOLUTION ‘D’ : 0.8ug(0.1%)
C : REF. SOLUTION ‘C’ : 1.6ug(0.2%)
B : REF. SOLUTION ‘B’ : 2.4ug(0.3%)
A : REF. SOLUTION ‘A’ : 8ug
 Impurity Quantification

Sample Std. Impurity

Spectral match of impurity in sample

m-chlorobenzoic acid in Bupropion HCl ANCHROM

Content Uniformity Test of Cinchocaine HCl

3D- display of 26 tracks

Cinchocaine Calibration curve

ANCHROM
COMPARISON
OF SAMPLES

ANCHROM
 TLC vs. HPTLC
DETECTION OF FALSE POSITIVE

Suspect

Sample Standard Spectra of std & sample at Cannot be visually

Rf 0.22 do not match! So they distinguished.
are different compounds.
The standard and one fraction of the sample have the same
Rf (0.22).By TLC, we would say that the impurity is present
but the TLC Scanner can record a UV abs spectrum in situ.
Here we see that the two spectra are different and hence
they are not the same compounds. Without the spectrum, a
false positive would have been reported!
ANCHROM
 Process Monitoring
Process Monitoring of Ethyl Carbazole

1. REACTION MASS AFTER 2 Hours 2. 4 Hours. 3. 6 Hours.

4. 8 Hours. 5. STD CARBAZOLE

ANCHROM
 Residue
Quant. of pesticide residues in grapes

Sample Std Carbaryl

Image (254 nm) Image (Vis)

ANCHROM
Quantification by Fluorescence
Aflatoxin B1 in fungal extract, ppb level

Fungal sample Aflatoxin B1

Image(366nm) ANCHROM
DRUGS FROM
BODY FLUIDS
Forensic drug screening
Morphine in urine

ANCHROM
BATCH TO BATCH CONSISTENCY
Polyherbal formulation

Batch Spectra

Image(254nm) Image(366nm)
ANCHROM
FERMENTATION
BROTH

Determination of rape seed oil in fermentation broth

ANCHROM
 RESIDUE ANALYSIS

SAMPLE RDX K6

HMX TNT TNAZ

ANCHROM
Spectra
EXPLOSIVES
 LIMIT TEST

Banned Amines – Limit : 30ppm

ANCHROM
 ASSAY

Known Impurity Spectrum of

Image at 254 nm
4-chloro-acetanilide 4-chloro-acetanilide

Quantification of 4- chloro- acetanilide from tablet

ANCHROM
MULTI DRUG FORMULATION

Image at 254 nm Spectral match of ambroxol HCl std. & sample

Cetrizine in sample Ambroxol in sample Spectra of cetrizine HCl std. & sample
ANCHROM
 IMPURITY PROFILE OF
NORGESTREL IN FORMULATION

Image (254 nm) Image (366 nm)

Norgestrel std (1%) Sample

ANCHROM
 HPTLC ANALYSIS OF
EZETIMIBE

Linearity Recovery & Label Claim

Ezetimibe Std Spectra Calibration

ANCHROM
DETECTION OF CORTICOSTEROIDS IN
FORMULATIONS

Std. Mix. Soln.

Image (254nm)

Spectra

ANCHROM
BANNED COLOURS IN
CHILLI POWDER

IMAGE (254nm)

IMAGE (Visible)

ANCHROM
 IDENTIFICATION OF E-123, E-124, E-129 IN
CARBONATED DRINK

Image (254) Image (visible)

std
std

sample sample

Spectra of allura red & sample at Rf 0.45 Spectra of Ponceau 4R & sample at Rf 0.19 Spectra of amaranth std

ANCHROM
FORMALDEHYDE IN MILK

Image (254) Milk Sample Dimedone

Milk sample + Dimedone Dimedone + Formaldehyde Milk Sample + Dimedone + Formaldehyde

ANCHROM
SHODAN IN AYURVEDA
Before Derivatization After Derivatization

Image at 254 nm Image at 36nm

ANCHROM
 GC - HPLC - HPTLC
GC HPLC HPTLC

¾ Solid stationary phase Liquid Solid

¾ Gaseous mobile phase Liquid Liquid
¾ Conditioning phase-nil nil Gas
¾ Samples should be volatile Non-volatile Non-volatile
¾ Sample invisible invisible visible during
Chromatography
¾ Closed system Closed system Open system
¾ Separating medium is
Tubular column Tubular column Planar column ( plate)
¾ 1 sample analysed at a time 1 sample upto 22 at a time / plate.
5 plate at a time
¾ Full automation Full automation Stepwise
Automation
¾ High temperature High pressure Room T & P

ANCHROM
 GC HPLC HPTLC

¾ Sample cleanup essential Essential Not essential

¾ Sample + stds Sequential Simultaneous
sequentially analysed analysis
¾ Reusable columns Reusable columns Disposable plates
¾ Unknown samples not No Yes
acceptable
¾ Preparative – Rare Not common Routine
¾ No absorbance spectrum Yes (PDA) Yes
¾ Post chrom. derivatisation
Very difficult Very difficult Easy
¾ Maintenance-Medium High Low
¾ Running cost – Medium High Low
¾ Samples/shift 5-25 5-25 Upto 100
¾ Analysis time
5-60 min/sample 5-60 min. 1 to 5 min / Sample

ANCHROM
 HPTLC
QUALIFICATION

1. INSTALLATION

2. OPERATION

3. PERFORMANCE
ANCHROM
 HPTLC
ELECTRONIC
SECURITY

21 CFR RULE 11

ANCHROM
 Anchrom Lab Activities
¾ Analysis
¾ Training
¾ Customer support
¾ Installation
¾ New method for India

ANCHROM
ANCHROM TEAM
PHOTO

ANCHROM
www.anchrom.com

ANCHROM
THANK YOU ! !

HPTLC SPECIALISTS