CAMAG TLC-MS INTERFACE 2 User manual

INSTRUCTION MANUAL
TLC-MS INTERFACE 2

Content
1Introduction................................................................................................................................................... 2
1.1 Precaution............................................................................................................................................... 2
1.2 Parts supplied.......................................................................................................................................... 3
2Unpacking/Installation.................................................................................................................................. 4
2.1 Unpacking............................................................................................................................................... 4
2.2 Installation environment .......................................................................................................................... 4
2.3 Installation .............................................................................................................................................. 4
2.4 Connection of the valve........................................................................................................................... 5
3Getting started .............................................................................................................................................. 6
3.1 Intended use ........................................................................................................................................... 6
3.2 Instrument .............................................................................................................................................. 6
3.3 Functional principle ................................................................................................................................. 7
3.4 Elution process ........................................................................................................................................ 9
3.5 Operation advice ................................................................................................................................... 10
4Maintenance and Service............................................................................................................................ 10
4.1 Cleaning................................................................................................................................................ 10
4.2 Decontamination................................................................................................................................... 10
4.3 User maintenance.................................................................................................................................. 11
4.4 Maintenance Data Sheet ....................................................................................................................... 11
4.5 Replacing of elution head ...................................................................................................................... 12
5Technical data.............................................................................................................................................. 12
6Application hints ......................................................................................................................................... 13
6.1 Practical procedure for elution ............................................................................................................... 13
6.2 Fittings .................................................................................................................................................. 15
6.3 Dead volumes ....................................................................................................................................... 17
6.4 System leaks.......................................................................................................................................... 18
Telltale signs of system leaks .......................................................................................................................... 18
6.5 Application example.............................................................................................................................. 19
Sample: diclofenac in methanol at a concentration of 0.113 mg/mL ............................................................... 19
Separation of Caffeine, Paracetamol, Acetylsalicylic Acid................................................................................. 20
Results ........................................................................................................................................................... 22
EC Declaration of Conformity

Introduction
Instruction Manual, Feb-17 2
1Introduction
1.1 Precaution
•Please read this operating manual before starting the installation! This manual
contains information and warnings the user has to follow to ensure reliable
operation of the instrument
•If the instrument is used in a manner not specified in this manual, the protection
provided by the equipment may be impaired
•This sign indicates (on instrument and in this manual) that failure to take note of
the accompanying information may result in damage of the instrument
•Attention: For safety reasons the instrument may only be used for the purposes
described in the operating manual
•To avoid injury use adequate safety equipment (protective goggles, gloves etc. if
applicable) when working with the instrument
•When working with the fluids of the instrument, be sure to take the appropriate
caution (protect your eyes from direct contact with liquid)
•The instrument may be used only by properly trained laboratory staff
•Use a damp lint free cloth for cleaning the instrument surface. Do not employ
aggressive detergents
•Only authorized personnel may open the instrument. Service and repair is only to
be performed by trained specialists. Use spare parts and consumables supplied by
CAMAG only. The warranty is voided if parts from other sources are used. Check
the service manual before you start service to reduce product-specific risks
•If the instrument is found to be defective, it must be switched off and steps must
be taken to ensure that it cannot be switched on by mistake
•If liquids penetrate the inside of the instrument, the power has to be disconnected
immediately. Small amounts of liquid can be wiped off and/or dried by means of a
hairdryer, with larger amounts of liquid a service technician has to be called. A test
of functionality has to be performed in all cases
•Carry out all safety checks and the preventive maintenance as recommended by the
manufacturer in order to assure your personal safety and the full functionality of
the instrument. Have an authorized service specialist perform any service not
described by this manual
•See original manufacturers' manuals for further safety data on third party
equipment supplied with the system
•The safety of any system incorporate with the equipment is the responsibility of the
assembler of the system

Introduction
Instruction Manual, Feb-17 3
•Lift/move/transport the system with the necessary care and with sufficient
manpower (install the transport security devices if applicable, transport it only in the
original packaging)
•This symbol indicates that this equipment must not be disposed of as unsorted
municipal waste but is to be collected separately as electrical and electronic
equipment (WEEE-Directive 2002/96/EC). To properly recycle the instrument or
parts of it you are requested to send the equipment back to the distributor,
producer or an adequate collection system at the end of its life. This will have
potential effects on the environment and human health
•Do not mix batteries -When replacing batteries, replace them at the same time with
new batteries of the same type
•Exhausted batteries should immediately be removed from the equipment and
disposed of accordingly. When discharged batteries are kept in the equipment for a
long time, electrolyte leakage may occur and cause damage to the instrument
•Laser (class 1) inside: Do not look directly into the laser beam
1.2 Parts supplied
Part no Description
140.0450-1
Pneumatic kit containing:
672.1016
Hose clip Ø 8-16mm
672.0029
Hose PVC
115.0050
Hose fitting
672.0080
Hose black
370.0002
2 Battery 1.5V ,AA size
022.8450
Colum Saver 2µm, SST
B.022.8440E
Instruction Manual
2 Allen key (for maintenance of 6 port Valve)
Accessories and spare parts
Part no Description
022.8446
Elution head for circular zones of 4mm diameter
022.8445
Elution head for oval zones of 2mm width
666.0001
10-32 (1/16”) RheFlex ® SS Fitting (nut and ferrule)
666.0002
Adapter -1/4-28 x 10-32 PEEK Interface 2
666.0003
Nanotight FEP tubing sleeve 1/16” OD
666.0004
Nanotight peek fitting (fingertight nut and ferrule)
666.4007
F-120 PEEK fitting
666.0017
Flangeless Nut 1/8"
666.0018
Flangeless Ferrule 1/8"
670.0030
Valve dummy connector (for protection)
022.8450
Colum Saver 2µm, SST
022.8420
Waste filter for cleaning sledge
660.0040
O-ring for waste container
Rotor seal for 6 port valve

Unpacking/Installation
Instruction Manual, Feb-17 4
2Unpacking/Installation
2.1 Unpacking
Observe the environmental requirements (2.1 Installation environment) when set-ting up
the instrument.
•From the upper compartment of the package carefully unpack all components and
accessories listed on the shipping list. Make sure the shipment is complete
•Carefully remove the instrument from the package and place it on a table
•Push the button (laser cross hair on/off) down and you will see laser crosshairs
on the positioning table. If there is no cross please check the batteries
2.2 Installation environment
The place for installation must meet the following requirements:
Connections
5 bar compressed air or N2. (backside of instrument)
Short capillaries from HPLC pump to TLC-MS interface 2 and
from there to the MS
Bench space
Width 27.5 cm; Depth 45 cm; Height 30 cm
Operating
temperature
The temperature should be within a range of 15 to 35 degrees
centigrade.
Humidity
Humidity and temperature conditions must not cause
condensation.
Additional inline
filter
CAMAG strongly recommends using an additional inline filter
between the TLC-MS Interface 2 (Valve port 1) and the MS.
The porosity of the filter is depending on the usage, CAMAG
recommends to use a filter with the porosity of 0.2 µm
2.3 Installation
•Make sure the batteries at the backside of the instrument are correctly inserted
•Connect your LC pump and MS system with the delivered set of 10-32 (1/16”)
RheFlex® Two-Piece fittings or with compatible alternatives.
oPort 2 from the LC pump
oPort 1 to the MS system
•Connect the black tube to your pressured gas supply system of 4 – 5 bar
compressed air or N2.
Your instrument is now ready to use!

Unpacking/Installation
Instruction Manual, Feb-17 5
2.4 Connection of the valve
Bypass position
Elution position
1
2
3
45
6
From LC pump
Compressed air
at cleaning
To inline filter and LCMS
Color
sleeve
Elution head
Internal
filter
5
Dummy plug
666.4008
Fig 1

Getting started
Instruction Manual, Feb-17 6
3Getting started
3.1 Intended use
The TLC-MS Interface 2 is intended for the use described in this manual.
3.2 Instrument
Fig. 2
Control elements:
1 Laser cross hair on/off
2 Piston up/down
3 Cleaning on/off
4 Screw for fixing the security cover
5 Piston with elution head
6 Positioning table
7 Lever in position elution/bypass
8 Air pressure adjustment
9 6-port valve for connection of capillaries from pump and to MS
10 Plate stopper

Getting started
Instruction Manual, Feb-17 7
3.3 Functional principle
The instrument extracts zones (round or oval) from TLC/HPTLC plates or aluminium foils
up to 20 x 20 cm. Plates can be positioned accurately and analysed zone by zone. Elutions
are possible with appropriate solvents, using the standard flow speed of the HPLC-MS
system (e.g. 0.1 ml/min). The system allows a semi-automatic operation involving piston
up/down movement, cleaning and backflushing of the elution head, manual positioning
and valve switching.
The elution head has two connections on the top side, one inlet and one outlet. On the
bottom surface there is a cutting edge seal with a height of about the thickness of the
plate layer. When the elution head is pressed onto a foil or glass plate the cutting edge
seal cuts into the adsorption layer and creates a leak-free seal.
In bypass position solvent flows directly to the MS-System. With the help of a laser
crosshairs the extractor head is easily positioned on a selected zone.
If the position of a zone is not visible, positioning is done with the help of rulers on the
instrument table.
Fig. 3: TLC-MS Interface 2 in bypass position
After lowering the piston, switch the valve to elution position. The solvent passes through
the elution head onto the silica gel, elutes the sample and transports it directly to the
LC/MS-System.
TLC/HPTLC plate or foil with separated substance zone
Force controlled
piston
in upper position
HPLC Pump
delivering
solvent
Laser projects a
crosshairs
Bypass
Frit

Getting started
Instruction Manual, Feb-17 8
Fig. 4: TLC-MS Interface 2 in elution position
After each elution remove the plate/foil. Make sure the valve is in position “Bypass” and
press the cleaning button at least 3 times for 3 seconds. The capillary from the plate to the
valve and internal frit are cleaned with the help of compressed air. At the same time the
cleaning sledge moves underneath the head to collect residuals of silica powder from the
elution head.
TLC/HPTLC plate or foil with separated substance zone
HPLC Pump
delivering
solvent
MS
System
Frit
Elution

Getting started
Instruction Manual, Feb-17 9
Fig. 5: TLC-MS Interface 2 in bypass position , cleaning active
3.4 Elution process
•Make sure, that the valve is in the rear, Bypass position
•Position your TLC plate/foil on the instrument table
•Press the laser cross hair button and you will see the laser cross hair on the plate. If
zones are visible you can easily position the TLC plate with the cross hair
•If zones are not visible you have to evaluate the position with a respective device
and position the plate with the help of the plate stopper and ruler on the
instrument table accordingly
•Toggle the Piston up/down switch to lower the elution head onto the plate/foil
•Switching the valve to front, Elution position will start the elution. An elution
time of about 1 minute should be sufficient (this is strongly depending on the flow
rate, solvent type and length of the capillaries)
•Switch the valve back in the rear, Bypass position
•Toggle the Piston up/down switch to bring the elution head into the upper position
•Continuously push the cleaning button at least 3 times for 3 seconds to clean the
elution head and capillaries.
TLC/HPTLC plate or foil with separated substance zone
Force controlled
piston
in upper position
HPLC Pump
delivering
solvent
Bypass
Frit
Cleaning
Pressure

Maintenance and Service
Instruction Manual, Feb-17 10
3.5 Operation advice
•Elution of a sample–free area on the plate cleans the system from residues of the
last elution. In this manner you can also measure the plate background for later
correction
•The positioning table must be wiped off before you put a plate or foil on it.
•Capillaries to the detector system should be kept as short as possible
•Increasing the flow rate results in faster elution time but larger elution volume and
a rise of pressure
•Caution: It is not recommended to leave the ports of the valve unconnected
without protecting them from dust. Please use the included taps or similar parts to
protect the ports
Check chapter Maintenance and Service for a trouble-free handling of the instrument.
4Maintenance and Service
4.1 Cleaning
Cleaning the instrument on a regular basis in an appropriate way assures problem-free
functionality over a long period.
A good way to perform a cleaning would be to follow the decontamination instruction
outlined below.
4.2 Decontamination
Before transportation or a longer term of not using your system, decontaminate it in an
appropriate manner. The decontamination procedure below reflects the minimal
requirements, so keep in mind that the procedure for your instrument has to be adapted
according the substances used.
Decontamination procedure:
•Disconnect the capillaries to the LC/MS system
•Fill a bottle with 70% Ethanol as cleaning solution and connect it to the pump. Adapt
the cleaning solution according to the used substances if necessary
•Place a clean plate underneath the elution head
•Perform 3 elution cycles (each 1 min). After each cycle, press the cleaning button 3 times
for 3 sec.
•Place a lint free cloth underneath the elution head
•Move the lever into position “Elution”
•Run the pump for 1 min
•Press the cleaning button 3 times for 3 seconds

Maintenance and Service
Instruction Manual, Feb-17 11
•Clean the instrument with a lint free cloth
4.3 User maintenance
Regular check/maintenance (at least every 3 months) by the user is strongly
recommended.
User maintenance procedure:
•Check the capillaries
•Check the 6 port valve for signs of leakage
•Clean the instrument with a lint free cloth
•Replace the respective parts as outlined in chapter “Maintenance Data Sheet”
4.4 Maintenance Data Sheet
Purpose
The maintenance data sheet informs about maintenance interval of the
respective instrument as well as the proposal for IQ/OQ interval if
applicable. In addition, it identifies parts subject to wear with the
respective replacement cycle and CAMAG order number.
Maintenance interval
Maintenance
12 month
IQ/OQ
12 month
Consumable parts
Part No.
Description
Replacement cycle
370.0002
2 Batteries 1.5V AA size
36 months/100h
022.8450
In-line frit
6 months/depends on usage
115.8448
Elution head, round 4mm h=
0.25mm
36 months/if leakage is
detected
115.8446
Elution head, oval 4x2 mm
h=0.25mm
36 months/if leakage is
detected
941.0040
Waste filter for cleaning sledge
36 months
660.0040
O-ring for waste container
36 months
Rotor seal for 6 port valve
36 months/if leakage is
detected or the pressure in
the system is too high

Technical data
Instruction Manual, Feb-17 12
4.5 Replacing of elution head
•Remove the security cover
•Remove the blue top cover
•Remove the elution head capillaries to port 3 and port 6 (the filter) of the valve
•Make sure the filter to port 6 of the valve is changed if the head is replaced
•Unfasten the elution head clip
•Remove the elution head carefully through the top of the instrument
•Insert the new head through the top of the instrument
•Fasten the elution head clip
•Mount the capillaries (color sleeve to the filter on port 6 of valve)
•Mount the blue top cover
•Mount the security cover
•Make an fake extraction to check if the elution head is properly positioned and no
leaks are visible
5Technical data
General data
Plate types
TLC/HPTLC glass plates and aluminium foils of sizes up to
20 x 20 cm and layer thickness up to 300µm
Standard flow rate
50 - 300 µL/min
Down force
400 N max
Gas supply pressure
4 – 6 bar compressed air or N2
Electrical data
Laser type
5 mW, laser class 1
Electrical power
Two batteries 1.5 V, AA/LR6 size
Battery duration
Up to 100 hours
Mechanical data
Dimensions
275 x 425 x 275 mm (w x d x h)
Weight
14.5 kg
Chemical data
Elution head capillaries
Stainless steel passivated and rinsed with reagent-grade
methanol
Frit
2µm stainless steel
Elution head
Stainless steel

Application hints
Instruction Manual, Feb-17 13
6Application hints
6.1 Practical procedure for elution
Installation
The TLC-MS Interface 2 can be connected to any brand of MS.
The type of MS is mainly defined by the tasks of your laboratory and by the availability.
For simple tasks, like confirmation of a structure or mass, an easy to use single quadrupol
MS is adequate. For more complex MS/MS tasks (or if it is the only available instrument) a
triple quadrupol MS, or an ion trap MS may be suitable. For identification of unknowns
even a time of flight (TOF), Q/TOF or an Orbitrap MS can be used.
Eluent delivery
For the delivery of the eluent, a HPLC pump is needed. You may use the internal pump of
the HPLC-MS. Remove the HPLC column if installed and connect the respective capillary
from the TLC-MS Interface 2 to the solvent delivery capillary of the HPLC (as described in
this manual).
You may alternatively use a stand-alone HPLC pump.
Connecting the TLC-MS Interface 2 to the MS
Connect the capillary from the HPLC pump to port 2 of the valve and port 1 directly to the
MS (it is recommended to bypass the UV detector cell, because it is sensitive to back
pressure from the TLC-MS Interface 2).
It is highly recommended to use an inlet filter before the MS. The inlet filter is a standard
feature of most MS. For more information check chapter 2.2 Installation environment.
Setting up the MS
Select the ionization mode according to substance requirements and availability: ESI, APCI
or APPI ionization, select appropriate mode (positive or negative).
Tune the MS according to the MS manufacturer’s recommendations.
Optimizing MS parameters
Ideally the substance to be investigated is available in pure form. If not, another substance
with similar properties can be used to verify the ionization conditions.
Eluents are often mixtures of acetonitrile or methanol with water. Mixtures of 80 %
organic phase with 20 % of water have been successfully used. Depending on the nature
of the substances to be investigated, protonation can be enforced by adding 0.1 % of
formic or acetic acid. Deprotonation can be enforced by adding 0.1 % of ammonia.
A flow of 0.2 mL/min is suitable for the elution.
Caution: Do not use any nonvolatile buffer salts (e.g. phosphates) in the eluent and use
only MS grade solvents!

Application hints
Instruction Manual, Feb-17 14
A general rule of thumb for HPLC-MS: Dilute the solutions you typically use for UV-
detection (approx. 1 mg/mL in e.g. methanol) by a factor of 1000 or 100, resulting in
concentrations of 1 to 10 µg/mL. Use flow injection analysis (FIA) of 0.1 µL and watch the
MS signal. If the MS signal is too small, inject 1 µL or more, until you get a clean mass
spectrum.
Fig. 6: Flow injection analysis FIA of 1 µL of 0.113 mg/mL diclofenac in methanol, upper
curve total ion current (TIC), lower curve single ion monitoring (SIM) of the masses
296.00, 297.00 and 298.00 dalton.
Fig. 7: Flow injection analysis FIA of 1 µL of 0.113 mg/mL diclofenac in methanol;
spectrum of diclofenac.
Use FIA to optimize MS parameters: Nebulizer pressure, drying gas temperature and
drying gas flow, capillary voltage and fragmentor voltage (consult the manufacturer’s
recommendations for your individual brand of HPLC-MS).
Optimizing TLC conditions

Application hints
Instruction Manual, Feb-17 15
If available use HPTLC plates. Prior to use, prewash plates by developing once to the upper
edge with neat methanol. Dry the plates for 10 min at 60°C in a vacuum drying oven or
by heating for 20 min at 120°C in a clean (!) convection oven.
The preferred sample application technique is the spray-on technique using the Automatic
TLC Sampler 4 or the Linomat 5. The spray-on technique allows application of relatively
large volumes in the form of narrow bands, resulting in a good resolution of the sample
zones after chromatography.
Use the sample solution with the concentration optimized by FIA experiments (usually 1 to
10 µg/mL). Apply a series of bands with increasing volume, e.g. 1, 2, 4, 8, 16 µL. The
selected band length depends on the type and the number of samples. For many different
samples on the same plate, a short band of 4 mm may be adequate. For fewer samples
which are to be analyzed more than once, a band length of 10 to 20 mm is preferred.
Alternatively samples can be applied with fixed volume capillaries of 0.5 to 5 µL. In this
case select a fixed volume and apply a series of individual dilutions.
Develop the plate in an appropriate device, e.g. the Automatic Developing Chamber
ADC2 or a Twin Trough Chamber, which allows for control of chamber saturation.
6.2 Fittings
In order to connect your tubing together and to connect it to the equipment in your HPLC
system; nuts, ferrules, and unions must be used. A ferrule is used to form the seal
between components. Utilizing the nut and ferrule combination, tubing is held in place by
a compression action.
Fig. 8: Fittings
Installing high pressure fittings
Before connecting fittings, ensure the end of the tubing has a clean, square, burr-free cut.
This is very important, as the square cut makes a flat surface for the tubing to contact the

Application hints
Instruction Manual, Feb-17 16
bottom of the receiving port. Improper cutting of the tubing could lead to dead volume,
resulting in peak distortion such as fronting, tailing or broadening.
Procedure with stainless steel nuts and ferrules:
Slide the fitting at least 5 mm from the end of the clean cutting tube.
•Insert the assembly into the receiving port, pushing the tubing into the port until it
bottoms out.
•Using the delivered wrench, tighten an additional
½ to 1 turn past finger-tight. Remove the fitting to
confirm the ferrule is swaged onto the tube.
•Because the ferrule is permanently attached to the
tubing we highly recommended that the fitting
only be used in the receiving port into which it was
initially swaged. Failure to this may result in dead
volume and/or leaks.
Fig. 9: Tubing sleeve
Making capillary tubing connections to 1/32”
capillaries
Smaller tubing sizes like 1/32” OD are being used with increasing frequency in
chromatography and related disciplines, and using these tubing sizes requires some special
skills normally unnecessary with 1/16” OD tubing. This is especially evident when using
the smaller tubing in receiving ports normally designed for the larger, 1/16” OD tubing.
Generally, the high pressure receiving ports in most of the equipment on the market have
an internal geometry supporting 1/16” OD tubing. This means that the port has an
internal pocket into which the 1/16” OD tubing extends. As long as tubing with the same
diameter
is used, potential dead volume resulting from the connection is kept to a minimum.
However, when the tubing is smaller than that which the receiving port was designed for,
it’s easy for dead volume to be introduced into the connection.
Numerous manufacturers have developed ways to adapt smaller, capillary tubing into the
receiving ports meant for larger OD tubing. Of the options available, two stand out as the
most popular. The first option involves customized ferrules which look similar to those
used for the larger tubing, but which feature smaller holes drilled through them to better
accommodate the capillary tubing.
Fortunately, the second option - the use of a special tubing sleeve - overcomes both of the
disadvantages exhibited by the customized ferrules. Tubing sleeves generally have a
controlled outer diameter (usually 1/16”) which allows them to fit into standard threaded
ports. And, since the sleeve will slide through a ferrule until it bottoms out in a receiving

Application hints
Instruction Manual, Feb-17 17
port, the tubing pocket beyond the ferrule will be completely filled to help avoid dead
volume in the connection.
Tubing sleeves also offer several side benefits. One is the structural support sleeves provide
to the outside of the capillary tubing, helping prevent damage to the tubing as it leaves
the fitting. Furthermore, the extrusion process generally allows a more concentric
connection than precision drilling and machining can provide, allowing for more accurate
tubing-to-thru-hole alignment and decreasing inline turbulence and mixing. For all these
reasons, using a tubing sleeve to connect capillary tubing into a receiving port is the
method of preference - unless the receiving port is specifically designed for use with the
size of tubing you are using!
6.3 Dead volumes
One of the concerns you will have as a chromatographer is about the dead volume
existing in your system. Dead volume is described as a small space within your HPLC
system where remixing of the sample may occur or where your sample can be diluted with
mobile phase. Dead volume can cause critical problems in your analysis of chemical
compounds, as it may bring about band broadening or split peaks in the end result,
making it difficult to obtain good data on your samples. You must keep this in mind when
making connections throughout your system.
Fig. 10: Potential dead volumes in capillary-based applications
The tube diameter is too small
Mixing chamber
The tube is
too short

Application hints
Instruction Manual, Feb-17 18
6.4 System leaks
Telltale signs of system leaks
Before you even see your first drip from a fitting, your system can tell you that the
problem exists. The most common signs of system leaks are as follows:
1. No flow or pressure
2. Pump pressures up, but no flow
3. Noisy baseline
4. Baseline drift
While all of these symptoms could also indicate problems completely unrelated to leaking
fittings, it is always easiest to start there. Not only are fitting leaks usually easy to repair,
they are also the least expensive part of the system that can cause problems.
Check to make sure your tubing is seated properly
When using finger tight fittings, the tubing must bottom out in the receiving port before
the nut and ferrule are tightened. If a gentle tug disengages your tubing after the fittings
have been tightened, loosen the fitting, push the tubing to the bottom of the receiving
port, and re-tighten the fitting.
The fitting may not be tightened enough
Stainless steel nuts and ferrules require a wrench to tighten them, even after repeated
use. Finger tight fittings also require a good turn; however, don’t use a wrench unless
instructed to do so, or you may damage the fitting.
Notice: Over tightening can cause leakage too!
You may be using incompatible fittings
Make sure you are using a nut and ferrule that are compatible with each other and with
the components of your system
Check the condition of the nut and ferrule
After repeated use, nuts (and especially ferrules) will gradually become de-formed to the
point of being incapable of creating the seal they were de-signed to make. Always keep
an extra supply of all the nuts and ferrules you are using so that you can replace them
quickly and avoid unnecessary down time.
Check the receiving port for damage
Sometimes a leaking connection has nothing at all to do with the nut and ferrule, but
with the receiving port. Ports that have had stainless steel fittings swaged into them are
especially susceptible to damage. Check the receiving port for visible burrs or scratches
and replace if necessary.
Evaluate chemical compatibility
Using fittings made of material that is incompatible with your mobile phase is a sure way
of creating leaks.

Application hints
Instruction Manual, Feb-17 19
6.5 Application example
Sample: diclofenac in methanol at a concentration of 0.113 mg/mL
Layer: 20x10 cm HPTLC plate Si60 F254, prewashed with methanol; UV 254 nm
Application: 1, 2, 4, and 10 μL as 8 mm bands
Mobile phase: toluene – ethyl acetate – acetic acid 50 : 40 : 1 (v/v/v)
Development: ADC2, saturated chamber, 70 mm from lower edge of plate
Under UV 254nm mark zones with a soft pencil.
Fig. 11: Different volumes of 0.113 mg/mL diclofenac in methanol ; 8 mm bands
MS measurement
Start the HPLC pump.
The eluent used in the presented example was 95% methanol – 5% water and 0.02%
formic acid, with a flow rate of 0.2 ml/min.
Start the MS for data acquisition only (no injection).
The following MS settings were used:
Source
APCI
Mode
positive
Capillary Voltage
4'000 V
Charging V
2'000 V
Corona Voltage
4'000 nA
Drying Gas Temp.
350°C
Vaporizer Temp
250°C
Nebulizer
5 l/min
Nebulizer Pressure
20 psig
Fragmentor
100 V
Mass Range
100 – 500 m/z
Scan
20 min