Basics of LC/MS

A Primer
Contents

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

Instrumentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

Interfacing LC and MS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Atmospheric Pressure Ionization . . . . . . . . . . . . . . . . . . . . . . . . . 4
API-Electrospray . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Atmospheric Pressure Chemical Ionization (APCI) . . . . . . . . . . 6
Scan and SIM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Collision-Induced Dissociation . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Adapting LC Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Molecular Weight Determination . . . . . . . . . . . . . . . . . . . . . . . . . 10

Combinatorial Chemistry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Pharmaceutical Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Biochemical Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Clinical Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Food Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Environmental Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Introduction
Liquid Chromatography/Mass Spectrometry (LC/MS) is fast becoming the preferred
tool of liquid chromatographers. It is a powerful analytical technique that combines
the resolving power of liquid chromatography with the detection specificity of mass
spectrometry. Liquid chromatography (LC) separates the sample components and
then introduces them to the mass spectrometer (MS). The MS creates and detects
charged ions. The LC/MS data may be used to provide information about the molecu-
lar weight, structure, identity and quantity of specific sample components.

Sample Types they impart little or no heat to the analyte

molecules, LC and LC/MS-based methods
LC/MS systems facilitate the analysis
can be applied to most organic compounds.
of samples that traditionally have been
Sample types range from small pharma-
difficult to analyze. Despite the power and
ceutical compounds to large proteins.
usefulness of gas chromatography/mass
spectrometry (GC/MS), many compounds Because it is a much more widely
are impossible to analyze with GC/MS. applicable method than GC/MS, LC/MS
is suitable for the analysis of large, polar,
LC/MS significantly expands the effective
ionic, thermally unstable and involatile
analytical use of mass spectrometry to a
compounds. Some of these compounds
much larger number of organic compounds.
can be made amenable to GC/MS by
Gas chromatography and GC/MS can be
derivatization, but LC/MS eliminates
used to analyze a small percentage of the
the need for time-consuming chemical
9 million registered compounds. Because
modifications. This permits MS analysis of
non-volatile, thermally
labile, or charged
100,000 molecules.
API-Electrospray
Molecular Weight

10,000

APCI Thermospray
1000 FAB
Particle
GC/MS Beam

nonpolar very polar

Analyte Polarity

Figure 1. Applications of various LC/MS techniques

2
Selectivity and Sensitivity
A mass spectrometer combined with a liquid
chromatograph can detect masses charac-
teristic of a compound or of a class of
compounds. The system can selectively
detect compounds of interest in a complex
matrix, thus making it easy to find and
identify suspected impurities at trace levels.
When configured to simultaneously detect
a range of masses (and depending on the
compound) LC/MS sensitivity can be com-
parable to that provided by a diode-array
detector (DAD). Far greater sensitivity is
possible when the LC/MS is configured to
detect only those masses characteristic of
the compounds being monitored.
Complementary Information
Using MS in combination with other LC
detectors gives richer information. For
example, a DAD acquires data on selected
ultraviolet (UV) and visible (Vis) wave-
lengths and spectra. This information is
useful for identifying unknown peaks and
1200000
800000 MS TIC
400000
4000
m/z
2000
648
12000
8000
m/z
4000
376
300
200 m/z
100 1325
5 10 15 20 25
for determining peak purity or for both.
An MS acquires mass information by
detecting ions; it offers molecular-weight
and structural information. The LC/MS
can be used with analytes that do not
have chromophores. The two orthogonal
sets of data can be used to confidently
identify, confirm, and quantitate
compounds. In addition, an LC/MS
can be used as a highly selective and
sensitive tunable detector. An MS
chromatogram for a single mass often
produces an interference-free signal that
offers high precision and low minimum
detection limits.
Using both a UV detector and a mass
selective detector is more effective
than using either one alone. There are
compounds (such as metabolites or
degradents) for which the UV-Vis spectra
of two analytes will be very similar and
it may be difficult to detect an impurity
based on UV spectra alone. It is also
possible to have impurities that have the
same mass, especially
at lower molecular
weights. It is rare,
however, for two
components to have
identical UV-Vis
spectra and mass.
Figure 2 shows the
ability to separate
polymer components
from an unresolved
peak using the
information available
in a mass spectrum.
This separation
would not be possible
using a conventional
UV detector.
30 35 40 min.

Figure 2. Separation of isomers in a chromatographically unresolved peak

3
Instrumentation
LC/MS systems have improved dramatically over the last 20 years. Instruments have
been transformed from complex, high-cost, highly advanced research tools to low-cost,
robust, easy-to-use routine detectors. And, as the instruments have been refined, more
applications have been developed.

Interfacing LC and MS Atmospheric Pressure

There has been a major focus on improving
the interface between the LC and the MS.
Liquid chromatography uses high pressure
to separate a liquid phase and produces a
high gas load. Mass spectrometry requires
a vacuum and a limited gas load. For
example, common flow from an LC is
1 ml/min of liquid which, when converted
to the gas phase, is 1 l/min. However, a
typical mass spectrometer can accept only
about 1 ml/min of gas. Furthermore, an
LC operates at near ambient temperature
where as an MS requires an elevated
temperature. There is no mass range
limitation for samples analyzed by the
LC but there are limitations for an MS
analyzer. Finally, LC can use inorganic
buffers and MS prefers volatile buffers.
Ionization
Atmospheric pressure ionization (API)
techniques are soft ionization processes
well suited for the analysis of large
and small, polar and nonpolar, labile
compounds. These techniques can be
used to rapidly confirm the identity of
a wide range of volatile and nonvolatile
compounds by providing sensitive
and accurate molecular-weight and
fragmentation information. API techniques
can be used in metabolite confirmation
analysis of most pharmaceutical
compounds, and other applications.

Recent developments in atmospheric

pressure ionization sources have expanded
the molecular weight, sample polarity,
and flow-rate limitation of older LC/MS
techniques. In many cases, analysts are
able to use unmodified high-pressure
LC methods.

4
API-Electrospray
Application
API-electrospray (API-ES) is useful in
analyzing samples that become multiply
charged such as proteins, peptides, and
oligonucleotides, as well as in analyzing
samples that are singly charged, such as
benzodiazepines and sulfated conjugates.
API-ES can be used to measure the
molecular weights of most polymers,
peptides, proteins, and oligonucleotides
up to 150,000 Daltons quickly and with
high mass accuracy. In biopharmaceutical
applications, chemists use API-ES to speed
protein characterization, to accurately
identify and characterize post-translational
modifications, and to quickly confirm the
molecular weight of synthetic peptides.
Process
API-ES is a process of ionization followed
by evaporation. It occurs in three basic
steps: (1) nebulization and charging;
(2) desolvation and;
(3) ion evaporation.
Nebulizer gas
Nebulization
The HPLC effluent
is pumped through a
Solvent spray
nebulizing needle which
surface of the liquid and forms a spray
of charged droplets. There is a concentric
flow of gas which assists in the nebuli-
zation process.
Desolvation
The charged droplets are attracted toward
the capillary sampling orifice. There is a
counterflow of heated nitrogen drying gas
which shrinks the droplets and carries
away the uncharged material.
Ionization
As the droplets shrink, they approach a
point where the electrostatic (coulombic)
forces exceed the cohesive forces. This
process continues until the analyte ions
are ultimately desorbed into the gas phase.
These gas-phase ions pass through the
capillary sampling orifice into the low-
pressure region of the ion source and the
mass analyzer, see Figure 3.
Electrospray ions
Heated nitrogen drying gas
-5,000 V
+

is at ground potential. +
+
+
+

+
+
+ + + + + + + + + + + + + + +

The spray goes through

a semi-cylindrical
electrode which is at Dielectric capillary entrance
a high potential. The
potential difference
Rayleigh Coulomb
between the needle and Evaporation limit
explosion
Evaporation Analyte ion
the electrode produces reached

++ +
+

a strong electrical field.

+ ---+

+ + + + -+ +
+ -+ +
+
+ -+ + ---+ +
+ -----++ ++---++
+-+ + + - - +
+ +- +
+ -- - + +++ + + + -+ +
+++ -----+ + -- - +
+
This field charges the + - -+ + + -+ + +++-- ---++
++- ++
++ -----+

+ -+ +++ +
+ -- ---++ + -+
++++ + +-- +
-+
+ --+-+ + -+ +++
+++
++--+
++ + +- + + -+-+
+ --+ + ++ +-+- +

Figure 3. API-electrospray ionization

5
Atmospheric Pressure Chemical
Ionization
Application
Atmospheric pressure chemical ionization
(APCI) is an ionization technique that is
applicable to a wide range of polar and
nonpolar analytes that have moderate
molecular weights.
Process
APCI, a process of evaporation followed by
ionization, is complementary to API-ES.
Nebulization and Desolvation
APCI nebulization is similar to that in
API-ES. However, APCI nebulization
occurs in a hot (typically 250°C–400°C)
vaporizer chamber. The heat rapidly
evaporates the spray droplets, resulting
in gas-phase HPLC solvent and analyte
molecules, see Figure 4.
Ionization
The gas-phase solvent molecules are
ionized by the discharge from a corona
needle. In APCI there is a charge transfer
from the ionized solvent reagent ions to
the analyte molecules in a way that is
similar to chemical ionization in GC/MS.
These analyte ions then are transported
through the ion optics to the filter and
detector.

HPLC inlet
Nebulizer gas
High wattage
heater Vaporizer

[Solv + H] + + A
Dielectric capillary
Solv + [A + H] +
+
+ +
+
+
+ + +
+ + +
+ +
+ +
+
+ +
+
+

+
+
+

Corona discharge needle

Figure 4. Atmospheric Pressure Chemical Ionization (APCI)

6
Scan and SIM
Mass spectrometers can be operated in
either a scan mode or a selected ion
monitoring (SIM) mode (Figure 5).
Scan Mode
In the scan mode, the instrument detects
signals over a mass range (e.g. from
50–2000 m/z) during a short period of time
(e.g. 2 sec). During this scan period, the
MS electronics sequentially read the
signals detected within narrower mass
intervals until the full mass range is
covered. The spectra that are stored
represent the detected signal for the
full mass range. Since full mass spectra
are recorded, this mode of operation is
typically selected for qualitative analysis,
or for quantitation when all analyte masses
are not known in advance.
Samples may be introduced into a mass
spectrometer by infusion or through an
HPLC. In the latter, it is important to match
the peak width and the scan range. The
1 Scan
Scan
abundance
m/z
time
SIM 1 Scan
abundance
m/z
time
Figure 5. Scan and SIM data acquisition
narrower the peaks, the shorter the total
scan time must be in order to get proper
peak definition. In order to get a short total
scan time, it may be necessary to reduce
the scan range.
SIM Mode
Mass spectrometers can also operate in
the selected ion monitoring (SIM) mode.
Rather than scanning continuously, they
can be set to only monitor a few mass-
to-charge ratios (m/z). As a result the
quadrupole is able to spend significantly
more time sampling each of the m/z values,
with a concomitant and large increase in
sensitivity. Moreover, because the cycle-
time between data points is often shorter
than it is in scan mode, quantitative
precision and accuracy are improved
through optimal peak-shape profiling.
Since the m/z values to be sampled must
be set in advance, SIM is most often used
for target compound analysis. For analyses
consisting of multiple target compounds,
SIM ion sampling choices can be time-
programmed to
Mass Range
match compound
elution time
windows. No data
is collected at m/z
values other than
those specifically
m/z sampled, so SIM
is rarely used in
Discrete Masses
qualitative analysis.
m/z
7
Collision-Induced Dissociation the parent ion is fragmented to a larger
degree. Electrospray CID has the
MS/MS is accomplished by a process advantage of more efficient transfer than
called collision-induced dissociation (CID) in a triple quadrupole.
in which ions break apart as a result
There are some limitations to using CID
of collisions with other molecules.
in a single quadrupole mass spectrometer.
Electrospray ionization can also be used
In a triple quadrupole or ion trap, a single
to produce CID spectra even with a single
ion can be selected and fragmented. In
quadrupole system. In many instances, a
CID with a single quadrupole instrument,
single quadrupole system can be used for
there will be multiple ions in the source
work that has traditionally required triple
that are fragmented. These other ions
quadrupoles or ion traps.
may interfere with the analyte of interest
Electrospray is a soft ionization technique by generating additional fragments. In
that produces a large number of molecular many cases, this problem can be solved
adduct ions. Adduct ions are typically by improving the chromatography to
protonated parent ions [M+H]+. These isolate the analyte (Figure 6).
ions are guided into the vacuum region by
applied voltages on lenses. By changing the
voltage, various degrees of fragmentation
may be achieved. With a low voltage, there
is little fragmentation; with higher voltages,

LC/MS
abundance

CID Q1
m/z

LC/MS/MS
abundance

Q1 Q2 (CID) Q3
m/z

Figure 6. CID with single quadrupole and triple quadrupole mass spectrometers

8
Adapting LC Methods
Compared to previous LC/MS interfaces,
the API-ES and APCI interfaces are
relatively rugged. In many cases, existing
methods may be used with little or no
adaptation. Some instruments allow flow
rates of 1 to 2 ml/min without splitting.
One of the most critical factors in adapting
LC methods is the choice of buffer.
Involatile buffers interfere with good
MS performance. For the best long-term
perfomance, it is highly recommended
that the method be modified to use a
volatile buffer.
HPLC inlet
Modern mass spec-
trometer designs Nebulizer
incorporate a number
of features to increase
their tolerance for
involatile buffers.
In the first-generation
+ +++ +
orthogonal to the lens axis and ions are
focused into the mass filter (Figure 7).
The extraneous material is pumped away.
Figure 8 shows a source that had been
subjected to 600 injections with a complex
involatile salt solution (approximately
9 g/l). Even after this abuse, the
performance was only slightly degraded.
This ruggedness makes this technique
attractive for laboratories in which there
are many users of the system and in which
large varieties of sample are commonly
handled. The instrument does not need to
be finely adjusted for every sample.
Figure 7.
Orthogonal
Skimmers spray of API-
Octopole
HED detector electrospray
source
Capillary
+ + + + + +

systems, the spray from + + +

the LC was directed

into the lens axis for
quadrupole systems.
In newer designs, the
Fragmentation
flow is directed zone (CID) Lenses Quadrupole

Figure 8. Salt deposits on an

HP 1100 Series LC/MSD System
API interface

9
Applications
LC/MS is suitable for many applications, from pharmaceutical development to envi-
ronmental analysis. The ability to detect a wide range of compounds has made API
techniques popular with scientists in a variety of fields.

Molecular Weight Determination is at the C-terminus where one peptide

One of the key applications of API-
electrospray and APCI LC/MS systems
is in determining molecular weights.
Differentiation of Similar Octapeptides
Figure 9 shows the spectra of two peptides
whose mass-to-charge ratio differ by only
1 amu. The only difference in the sequence
has threonine and the other has threonine
amide. The smaller fragements are
identical in the two spectra, indicating
that large portions of the two peptides
are similar. The larger fragments contain
the differentiating peptides.

100
Ala-Ser-Thr-Thr-Thr-Asn-Tyr-Thr 858.5

80

60

40
739.3

880.3
444.2

576.3
498.2

841.3
343.1

397.2

20

0
857.5

100 Ala-Ser-Thr-Thr-Thr-Asn-Tyr-Thr amide

80

60
879.5

40
739.3

840.3
576.3
444.2
379.1

20

400 600 800 m/z

Figure 9. Octapeptides with 1 amu difference

10
Determining the Molecular Weight of spectrometer measures the mass-to-charge
Green Fluorescent Protein ratio, the molecular weight of a large
molecule such as GFP can be determined
API-electrospray LC/MS can be used to
if it is multiply charged.
rapidly determine the molecular weight
of a protein. Although pure proteins can The upper part of the display in Figure 10
be done by infusion, this recombinant shows the mass spectrum of the chromato-
protein required chromatography. Green graphic peak. There is a regular pattern
flourescent protein (GFP) is a 27,000 to the spectral peaks, each one of which
Dalton polymer with 238 amino acids. It represents the molecule with a different
emits a green light when excited by UV. number of charges. The lower display
is a deconvoluted spectrum generated
Even though GFP is a very large molecule,
by the data system for the singly
a mass spectrometer with a smaller mass
charged molecule.
range can be used to determine its
molecular weight. Since the mass

Figure 10. Molecular weight

determination of
789.95
839.45
895.45 866.45

Green Fluorescent Protein

120000 by API-electrospray
767.55
746.15

100000
926.15
959.15

80000
726.25

1032.95
994.85

1074.15

60000
1118.85
1167.55
1220.45
1278.75

40000
1342.65

1413.25

1579.65

20000

1000 1500

1000000

800000
Deconvoluted spectrum
600000 MW = 26828.84
400000

200000

26700 26800 26900 27000

11
Combinatorial Chemistry
Combinatorial chemistry has transformed
the drug discovery process. Rather than
creating compounds through manual
manipulations, robotics is applied to
generate whole classes of compounds from
sets of reagents. Reactions are typically
done using 96-well capacity (or greater)
plates. These plates are loaded into an
autosampler that injects samples into the
LC/MS. The instrument acquires the data
and produces a report showing whether
the compounds detected are of the
expected molecular weight.
12
Recent advances in software allow many
chemists to use the high sample capacity
of automated LC/MS systems. Trays are
loaded on the autosampler; the method is
specified; the instrument analyzes each
well in the plate; and the data system
prints a report. Figure 11 shows a typical
display of a sample tray. Green dots show
the locations where the reaction product
is of the expected molecular weight. Red
dots show where the reaction product did
not have the expected molecular weight.
A more detailed display and report of each
position is possible.
Figure 11. Display for
combinatorial chemistry
on the HP 1100 Series
LC/MS System
Pharmaceutical Applications a series of benzodiazepines was run using
both UV and MS detectors. The UV trace
Rapid Chromatography could not be used for quantitation, but the
of Benzodiazepines extracted ion chromatograms for the MS
could be used.
The information available in a mass
spectrum allows some compounds to be The mass spectral information provides
separated even though they are chroma- additional confirmation of the identity.
tographically unresolved. In this example, Chlorine has a characteristic pattern
because of the relative
abundance of the
mAU Rapid chromatography and isotopic information
2 UV two most common
1.5 isotopes. In Figure 12,
1 343.1
the triazolam spectrum
0.5 Triazolam
345.1 shows that the molecule
0
2 Cl has two chlorines,
350
1 2 3 4 min and the diazepam
MS spectrum shows that
300
Triazolam 285.2
it has only one.
250
Clobazam
Diazepam
200
Diazepam
Detection of
287.2
150 Alprazolam 1 Cl Degradation Products
100 Estrazolam for Salbutamol
Lorazepam 250 300 350
50 Detecting degradation
oxazepam Diazepam
0 products can often be
1 2 3 4 min difficult because they
can be structurally very
Figure 12. Benzodiazepines by API-ES similar to the original
product. If the chromo-
Norm. UV Signal and spectra MS TIC and EICs
phoric region is intact,
160 the two compounds
120 120 cannot be distinguished
80
with a UV detector.
100 m/z 346
40
m/z 282 The UV spectra for the
80 m/z 224 salbutamol and its
200 250 300 350 nm MS TIC degradation products
60
mAU are very similar. The
20
40
unique mass spectral
15
* fragments can be used
10 *
5 * * * 20 to identify the com-
*
0 pounds. Figure 13
0 shows the extracted
5 10 15 20 25 30 min ion chromatograms
2 4 6 8 10 12 14 16 min
for various masses.

Figure 13. Salbutamol degradation products

13
Biochemical Applications
Detection of Glycosylation in a Tryptic
Map of a 60 kD Protein
Proteins are sometimes chemically or
enzymatically digested to identify the
components. The digestion mixture is then
chromatographed to produce a peptide
map. Figure 14 shows two chromatograms
for a 60,000 Dalton glycoprotein from a
mass spectrometer and a UV detector.
Although the chromatographic patterns
with the two detectors are very similar,
the mass spectrometer can provide
additional data. Glycosylation is the
covalent modification of certain amino
acids which result in the attachment of
complex carbohydrates. The sugars can
be fragmented from the peptide fragments
using in-source collision-induced
dissociation (CID). The code at the right
in Figure 15 is used to designate various
markers. Since glactose and mannose have
the same molecular weight, they cannot be
distinguished from each other. Therefore,
they are designated simply as Hex.
Figure 16 shows the extracted ion
chromatogram (EIC) for three of the
glycosylation markers. The fragments with
the glycosylation markers can be easily
identified.

Abundance MS TIC
36 pmol
2000000

1500000

1000000

500000

mAU
20 UV @ 214nm
15

10

15 20 25 30 35 40 45 min

Figure 14. Tryptic map of a 60 kD protein

14
 Fucose Mass: 146
Residue Marker Ion: 147

Sialic Acid Mass: 291

Residue Marker Ion: 292

HexNAc Mass: 203

Residue Marker Ion: 204

Galactose Mass: 162

Residue Marker Ion: 163

Mannose Mass: 162

Residue Marker Ion: 163

Figure 15. Typical glycosylation markers

Total Ion Current

20000000
10000000

m/z 204: HexNAc Fragmentor @ 300 V

150000

50000

m/z 292: Sialic Acid

80000

40000

m/z 366: HexNAc + Hex

150000

50000

15 20 25 30 35 40 45 50 55 min

Figure 16. Detection of glycosylation

15
Clinical Applications Food Applications

High Sensitivity Detection of Aflatoxins are toxic metabolites produced

Trimipramine and Thioridazine by certain fungi in foods. Figure 18
shows the total ion chromatogram for
For some compounds, MS provides more
four aflatoxins; each could be uniquely
sensitive detection. Trimipramine is a
identified by their mass spectra
tricyclic antidepressant with sedative
(Figure 19).
properties. Thioridazine is a tranquilizer.
Figure 17 shows these compounds in a
urine extract at a level that could not be
detected by UV. To get the maximum
sensitivity, the analysis was done by
selected ion monitoring.

mAU
UV
200
280 nm
0

80000
Trimipramine EIC
m/z 295
0

40000 Trimipramine EIC

m/z 100
0

20000
EIC Thioridazine
m/z 371
0

8000
EIC Thioridazine
m/z 126
0
1 2 3 4 5 6 min

Figure 17. Trimipramine and thioridazine in urine extract

16
 Scan of 2 ng
4
TIC
200000

160000
1) Aflatoxin G2 3
2
120000 2) Aflatoxin G1
1

3) Aflatoxin B2
80000
4) Aflatoxin B1

40000

6.00 8.00 10.00 12.00 14.00 16.00 18.00

Figure 18. Afltoxins by API-ES

315
+ +
331 [M+H] [M+H]
90 Aflatoxin G2 90 Aflatoxin B2
O O 313 O O
+
O O [M-OH] O
50 50
353
O O
OCH 3
+ 337
[M+Na] O O OCH3 287 +
[M+Na]

120 160 200 240 280 320 360 120 160 200 240 280 320

329 313
[M+H] + [M+H] +
90 Aflatoxin G1 311 90 Aflatoxin B1
O O +
[M-OH] O O
O O
50 50 O
351 285 335
O O OCH3 243
283 [M+Na]+ O O OCH3 [M+Na]
+

120 160 200 240 280 320 360 120 160 200 240 280 320

Figure 19. Mass spectra of aflatoxins

17
Environmental Applications derivativation and fluorescence detection.
The chromatogram in Figure 20 is
Carbamates by APCI-LC/MS that of a tomato extract that has been
spiked with 11 carbamates. Using APCI,
Carbamates are a class of pesticides these compounds are detected without
usually analyzed by post-column derivitization. The extracted ion chromato-
gram displays the peaks
41.5 pg/component
41.5 pg/component for the individual
compounds.
400
MS TIC
200 Detection of
Phenylurea
Herbicides
40 Aldicarb Sulfoxide m/z 132
20 Many of the phenylurea
herbicides are very
300 Carbofuran m/z 222 similar and difficult to
150
distinguish with a UV
detector (Figure 21).
40 Carbaryl m/z 202
20 Monuron and diuron
have one ring and differ
40 Promecarb m/z 226 by a single chlorine.
20 Chloroxuron has two
2.5 5 7.5 10 12.5 15 17.5 min chlorines and a second
benzene ring attached
Figure 20. Carbamates by APCI to the first by an
oxygen. The UV-Vis
UV shows class; MS identifies species 233.1 spectra are similar for
Norm. Diuron diuron and monuron,
500
400 199.2
but different for
300 Monuron
chloroxuron. Each
200 of these compounds
100 291.2 has a distinct mass
0 Chloroxuron
spectrum. The standards
200 250 300 350 nm
100 200 300 400 500 were run with an
*
API-electrospray LC/MS
mAU
100 system.
80
* *
60

40
20

5 10 15 20 25 min

Figure 21. Phenylurea herbicides by API-ES

18
19
Notes

20
21

The HP LC/MSD system has
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Information, descriptions and
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Hewlett-Packard Company
All Rights Reserved.
Reproduction, adaption or translation
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