Multi-Residue Pesticide Screening

and Quantitation in Difficult Food

Matrixes Using the Agilent 6495 Triple
Quadrupole Mass Spectrometer

Application Note

Authors Abstract
Dan-Hui Dorothy Yang, Anabel Fandino, This Application Note describes a UHPLC/MS/MS-based multi-residue method
and Na Pi for the determination of more than 250 pesticides and pesticide metabolites in
Agilent Technologies, Inc. food samples. The method benefits from the increased chromatographic resolution
Santa Clara, CA of the Agilent 1290 Infinity UHPLC System, the versatile ionization capabilities
of the Agilent Jet Stream ionization source, and the innate sensitivity of the
Thomas Glauner
Agilent 6495 Triple Quadrupole LC/MS System. The method has been applied to
Agilent Technologies, Inc.
the analysis of pesticide residues in complex matrixes such as black tea. Matrix
Waldbronn, Germany
effects in the ionization were controlled by extensive dilution of the sample
extracts prior to injection.

Our results demonstrate that the increased sensitivity of the 6495 Triple
Quadrupole LC/MS System enables the accurate quantitation of targeted
pesticides below the maximum residue limits (MRLs) specified by the European
Commission, most of them even in the 1:100 diluted extracts, with improved
precision and excellent robustness.
Introduction below the MRLs stipulated by the EC. precision of the analytical method is
Furthermore, the extensive dilution demonstrated for diluted black tea
The screening and quantitation of
of sample extracts requires very high samples.
pesticide residues in food products is one
precision, as even small deviations
of the most important and demanding
applications in food safety. There are
result in considerable inaccuracies when Experimental
multiplied with high dilution factors.
more than 1,000 pesticides and pesticide
Reagents and chemicals
metabolites that can be present in food This Application Note shows the
and, thus, are regulated and controlled. All reagents and solvents were HPLC or
development of an UHPLC/MS/MS
The European Commission regulation (EC) LC/MS grade. Acetonitrile and methanol
method for the screening and quantitation
396/2005 and its annexes set maximum were purchased from Honeywell
of hundreds of pesticides in food samples.
residue limits (MRLs) for more than (Morristown, NJ, USA). Ultrapure water
The method was developed using the
170,000 matrix‑pesticide combinations was produced using a Milli-Q Integral
Pesticide tMRM LC/MS Application
for food1. Similar regulations are in place system equipped with a LC-Pak Polisher
Kit (p/n G1733BA). Transitions for
in other regions. Most pesticides are and a 0.22-µm point-of-use membrane
all compounds in the comprehensive
analyzed with multi‑residue methods filter cartridge (EMD Millipore, Billerica,
pesticide standard mix (p/n 5190-0551)
covering hundreds of compounds, MA, USA). Formic acid was from Fluka
and a few additional pesticides of interest
which are applied to various food (Sigma-Aldrich Corp., St. Louis, MO, USA)
were included in the method. An Agilent
commodities for both screening and and ammonium formate solution (5 M)
1290 Infinity UHPLC System was coupled
quantitation. Therefore, fast and reliable was from Agilent (p/n G1946‑85021).
to the highly sensitive Agilent 6495 Triple
analytical methods are required to Pesticides were included in the Agilent
Quadrupole LC/MS System operated with
allow identification and quantitation comprehensive pesticide mixture
dynamic MRM mode with fast polarity
of hundreds of pesticides at low (p/n 5190-0551). A limited number of
switching. Several modifications to the
concentrations in a broad range of food additional pesticides were purchased
previous high-end triple quadrupole mass
matrixes with confidence. Criteria for the from Fluka (Sigma-Aldrich Corp., St. Louis,
spectrometer design resulted in higher
identification of pesticide residues and MO, USA). Immediately before use, the
analytical performance.
requirements for method validation and eight submixes of the comprehensive
quality control procedures for quantitation • New mass filter one (MS1) ion pesticide mixture and the mixed stock
are specified in guidance documents such optics for increased precursor ion solution of the additional pesticides
as SANCO/12571/20132. were combined and further diluted
transmission
with acetonitrile to a final pesticide
Matrix effects in electrospray ionization, working solution containing more than
which change considerably between • An improved curved and tapered 250 pesticides at a concentration of
different food samples, present a collision cell providing enhanced 10 µg/mL. This solution was used for
significant challenge to the accurate MS/MS spectral fidelity spiking the QuEChERS extracts and
quantitation of pesticides. There are for the preparation of the calibration
different strategies to compensate for • A new ion detector operating at samples. Eight calibration samples
matrix effects such as matrix matched dynode accelerating voltages of up with concentrations ranging from 0.02
calibrations, standard addition, or the to 20 kV to 100 ng/mL were prepared in pure
use of internal standards. However, acetonitrile.
matrix matched calibrations do not • A new autotune optimized for
fully compensate for variations in Sample preparation
speed and sensitivity
matrix effects within a commodity or
Tea, orange, and tomato samples were
a commodity group. Standard addition
In addition, the 6495 Triple Quadrupole obtained from a local grocery store.
requires multiple injections for each
LC/MS System uses the proven Agilent Samples were extracted according to
sample, which reduces productivity.
Jet Stream Ionization source and the dual the citrate buffered QuEChERS protocol
The use of isotopically labelled internal
stage ion funnel. Enhanced sensitivity using Agilent BondElut QuEChERS
standards is probably the most attractive
gives enhanced peak area response and kits (p/n 5982-5650). Ten grams of
approach, however, it is not applicable for
improved peak area precision, which homogenized fruit and vegetable or
all target compounds in a multi-residue
ultimately leads to lower detection limits 2 g of tea were weighed into 50-mL
pesticide method. Sample dilution is
compared to previous designs. The polypropylene tubes and extracted
another approach to minimize matrix
enhanced sensitivity was used for the with 10 mL acetonitrile for 1 minute
effects3 but requires the use of highly
extensive dilution of complex food sample while shaking vigorously by hand. The
sensitive analytical instruments due
extracts to minimize matrix effects in the tea samples were wetted with 8 mL
to the need to detect contaminants
electrospray ionization. The improved ultrapure water for 2 hours prior to
extraction. Raw extracts were cleaned
up by dispersive SPE using primary

2
secondary amine (PSA, p/n 5982‑5256). Data were evaluated using the Agilent minimum amount of analyte required
In black tea samples, graphitized carbon MassHunter Quantitative Analysis to produce a signal that is statistically
black (GCB) contained in the Agilent Software. Calibration was done using distinguishable from background noise
BondElut QuEChERS EN dispersive neat standard solutions and linear, with a confidence level of 99 %. This
SPE tubes (p/n 5982-5356H) was also 1/x weighted calibration curves. The approach has much more relevance for
used for cleanup. Final extracts of blank lower limits of quantitation (LLOQs) routine operation as it avoids ambiguity
samples were spiked in five relevant correlate with the instrument detection related to the variation in the chemical
concentrations with the comprehensive limits (IDLs) in black tea matrix. IDL is noise and subjectivity in the way
pesticide working solution and then calculated based on the relative standard signal‑to-noise (S/N) is determined4. In
diluted 1:5, 1:10, 1:20, 1:50, and 1:100 with deviation of a series of replicates of addition, it is directly correlated to the
acetonitrile. Matrix matched standards a low level sample that is not higher precision of the analytical method, which
and dilutions were prepared immediately in concentration as 2 to 5 times the is important when doing an extensive
before injection, and were measured with detection limit. The IDL is defined as the dilution of sample extracts.
five technical replicates.
Table 1. Instrument parameters.
Equipment
Agilent 1290 Infinity UHPLC System
Separation was carried out using an
Column Agilent ZORBAX RRHD Eclipse Plus C18, 2.1 × 150 mm, 1.8 µm
Agilent 1290 Infinity UHPLC system
(p/n 959759-902)
consisting of an Agilent 1290 Infinity
Column temperature 40 °C
Binary Pump (G4220A), an Agilent 1290
Infinity High Performance Autosampler Injection volume 2 µL
(G4226A), a sample cooler (G1330B), and Speed Draw 100 µL/min, Eject 200 µL/min
an Agilent 1290 Infinity Thermostatted Autosampler temp 6 °C
Column compartment (G1316C). The Needle wash 10 s with acetonitrile/water (50/50; v/v)
UHPLC system was coupled to an Agilent Mobile phase A) 5 mM ammonium formate + 0.1 % formic acid
G6495 Triple Quadrupole LC/MS System B) 5 mM ammonium formate + 0.1 % formic acid in methanol
equipped with an Agilent Jet Stream Flow rate 0.4 mL/min
electrospray ionization source. Agilent Gradient program Time B %
MassHunter Workstation Software was 0 5
used for data acquisition and analysis 0.5 5
(v. B.07.00). 3.5 50
17.0 100
Method 20.0 100
20.1 5
The 1290 Infinity UHPLC System
Stop time 20.1 minutes
conditions are summarized in Table 1, and
a summary of the 6495 Triple Quadrupole Post time 3 minutes
parameters are shown in Table 2. Agilent 6495 Triple Quadrupole LC/MS System
Identification of polarity, precursor and Ion mode Positive and negative ESI with Agilent Jet Stream
product ions, as well as optimization Scan type Dynamic MRM
of collision energies, was taken from Drying gas temperature 120 °C
the Agilent Pesticide tMRM LC/MS Drying gas flow 17 L/min
Application Kit, and was further optimized
Sheath gas temperature 300 °C
using Agilent MassHunter Optimizer
Software. Analysis was carried out Sheath gas flow 12 L/min
with positive and negative electrospray Nebulizer pressure 30 psi
ionization in dynamic multiple reaction Capillary voltage 3,500 (pos/neg)
monitoring (dMRM) in a single analytical Nozzle voltage 300 V (pos); 500 V (neg)
run. A 2-μL amount of the final extract Cycle time 500 msec
was injected into the UHPLC/MS/MS. Total number of MRMs 532 (positive: 509/negative: 23)
Maximum number of 68
concurrent MRMs
Minimum dwell time 5.10 ms
Maximum dwell time 249.09 ms
MS1 and MS2 resolution Unit

3
 Results and Discussion parameters such as collision energy Figure 1 shows the chromatogram of
and cell acceleration voltage were fine a tea extract spiked with more than
Development of the optimized but only minor deviation from 250 pesticides at a concentration
UHPLC/MS/MS method optimized values for previous models was of 10 µg/kg, and diluted 1:20 with
The pesticide screening method observed. The prefilter and the detector acetonitrile prior to injection.
developed for the Agilent Pesticide tMRM were adjusted according to mass during
LC/MS Application Kit was transferred the instrument’s autotune. Sheath gas
to the 6495 Triple Quadrupole LC/MS temperature was optimized using the
System. The method was extended MassHunter Source Optimizer Software
to include several relevant acidic to produce the highest abundance for
herbicides, and fast polarity switching the majority of target compounds, and to
was employed. Compound-dependent not compromise labile and ammonium
adduct-forming compounds.

×10 4

Fenazaquin
2
1.9
1.8
1.7
1.6
DEET

1.5
1.4
Pyracarbolid

1.3
Pirimicarb

Methabenzthiazuron
Prometon

1.2
Furalaxyl

Pyriproxyfen
Thiacloprid
Tricyclazole
Acetamiprid
Counts

1.1
Pencycuron
Thiabendazole

Carbofuran

1
Isoxaben

Beflubutamid
Triazophos

Prosulfocarb

0.9
Clomazone
Aminocarb

0.8
Metolachlor

Triflumizole

Etofenprox
Benalaxyl

0.7
0.6
Picolinafen

Carbosulfan
Spiromesifen

0.5

Ivermectin B1a
0.4
Oxamyl

0.3
0.2
0.1
0
2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 8.5 9 9.5 10 10.5 11 11.5 12 12.5 13 13.5 14 14.5 15 15.5 16 16.5 17
Acquisition time (min)

Figure 1. Chromatograms of more than 250 pesticides spiked into black tea at the MRL and diluted 1:20 with acetonitrile (corresponding to a concentration of
0.1 ng/mL). For the sake of clarity, only part of the chromatographic peaks are labelled.

4
Evaluation of increased These enhancements ultimately lowered lower RSD values for most pesticides
instrument performance detection limits compared to previous at the same dilution levels. The relative
designs. The empirical observation that standard deviation (RSD) of a series of
The MS1 ion optics has demonstrated
supports this hypothesis is shown in replicates at a low concentration level is
a noticeable increase in the precursor
Figure 2, which compares the obtained a universal measure of the ion efficiency,
ion transmission, and an increase up
area RSDs on a 6495 Triple Quadrupole and can be used for the estimation of
to a factor of 3 has been observed,
System versus a 6490 Triple Quadrupole the quantitation limits. A low RSD value
depending on the ion mass. In addition,
for 50 pesticides spiked into black tea at has much more relevance than S/N
the detector design results in signal gains
the MRL and diluted in different ratios measurements, which can change based
especially for large fragment ions and
with acetonitrile. The selection of these on the selected noise region and the
negative ions across a broad mass range.
50 pesticides was based on relevance. software algorithm used for calculation.
When comparing the area response of
Several of those compounds were found A particular RSD can be defined as the
pesticides acquired with the 6495 Triple
in official control samples above the minimum amount that can be reliably
Quadrupole System to results from the
MRL and thus, the import of tea into the detected, as long as the noise level does
earlier model, a compound dependent
European Union was blocked. The area of not significantly contribute to RSD values.
area gain of up to a factor of 5 was
the blue polygon is considerably smaller For pesticide residues, a maximum
observed.
than the area of the red polygon, which RSD of 20 % has been specified as the
Enhanced ion transmission not only indicates that the improved ion statistics minimum performance requirement in
resulted in increased peak areas but of the 6495 Triple Quadrupole System SANCO/12571/2013.
also in improved peak area precision. instrument translates into considerably

Acephate (1 in 100)
Triazophos (1 in 100) Acetamiprid (1 in 100)
Thiamethoxam (1 in 100) Alanycarb (1 in 50)
25.0
Thiacloprid (1 in 100) Aldicarb (1 in 50)
Thiabendazol (1 in 100) Azaconazole (1 in 50)
Tebufenozid (1 in 50) Azinphos -ethyl (1 in 20)
20.0
Tebuconazole (1 in 50) Azinphos -methyl (1 in 20)

Spinosyn A (1 in 50) Azoxystrobin (1 in 100)

15.0
Pyridaben (1 in 100) Benalaxyl (1 in 100)

Pirimicarb (1 in 100) Bifenthrin (1 in 100)

10.0
Phosalone (1 in 50) Buprofezin (1 in 100)

Oxamyl (1 in 100) 5.0 Butocarboxim (1 in 20)

Nitenpyram (1 in 20) Carbaryl (1 in 50)

0.0
Monocrotophos (1 in 100) Carbendazim (1 in 100)

Methomyl (1 in 50) Chlorantraniliprole (1 in 20)

Methidathion (1 in 20) Chloroxuron (1 in 100)

Methamidophos (1 in 100) Chlorpyriphos (1 in 50)

Metamitron (1 in 100) Cyazofamid (1 in 50)

Isocarbophos (1 in 100) Cycluron (1 in 100)

Imidacloprid (1 in 20) Desmedipham (1 in 100)

Imazalil (1 in 20) Diethofencarb (1 in 100)
Fuberidazol (1 in 100) Difenoconazole (1 in 50)
Flufenoxuron (1 in 100) Dimethoate (1 in 100)
Ethion (1 in 100) Dimethomorph (1 in 20)
Diuron (1 in 100) Dimoxystrobin (1 in 100) 6490 (RSD in %, n = 5)
Diniconazole (1 in 20) 6495 (RSD in %, n = 5)

Figure 2. Comparison of area RSDs for pesticides spiked into black tea at the MRL and diluted in different ratios with acetonitrile for the Agilent 6495 LC/MS
(blue) and the Agilent 6490 Triple Quadrupole (red).

5
With the updated design of the 6495
Triple Quadrupole LC/MS System, 3
more pesticides can be detected at low
concentrations in QuEChERS extracts of 14
different food commodities according to 22
the quality criteria specified in the SANCO
guidelines. In tomato and orange extracts,
all pesticides were easily detected at the 1:5 dilution
lowest spiked concentrations of 1 ng/g. 1:10 dilution
45
The tea matrix at this concentration has
a slightly smaller detection rate due to 1:20 dilution
170
the 5-fold lower sample amount and the 1:50 dilution
more complex matrix. Figure 3 shows
the detection rate of the pesticides for 1:100 dilution
different dilution levels in the black tea
matrix.

Under the applied experimental

conditions, approximately 67 % of all the
spiked pesticides were easily detected Figure 3. Pesticides spiked in black tea extract to 2 ng/mL and diluted at different levels. 170 pesticides
with an RSD below 20 % in the 1:100 can be detected at the 1:100 dilution level with an area RSD < 20 %. Additional compounds are detected
dilution, corresponding to a concentration at higher concentrations, that is, at lower dilution levels.
of 0.02 ng/mL. In addition, approximately
20 % were detected with acceptable
precision in the 1:50 dilution, and another
~10 % in the 1:20 dilution. Excellent
precision was observed for replicate
injections of these samples within a
72‑hour worklist.

Minimizing matrix effects by Figure 4 shows the chromatograms of While a weak matrix effect was observed
alanycarb and oxamyl spiked in black tea for diuron and flufenoxuron, the signal
dilution of sample extracts suppression for monocrotophos and
extract corresponding to 10 µg/kg and
The ability to extensively dilute sample diluted with acetonitrile prior to injection alanycarb in the nondiluted tea was
extracts to remove matrix effects is an in different dilution ratios. substantial. However, when diluting the
attractive capability to many routine final extract 1:10, more than half of the
testing labs. It enables quantitation Upon 1:5 dilution, the signal for alanycarb compounds showed adequate recoveries
of complex samples against a solvent increased. For oxamyl and the further of over 70 %. Very few compounds
calibration. A possible cause for matrix dilution levels of alanycarb, the peak required a larger dilution of 1:50, or even
effects in electrospray ionization is the areas decreased less than the extent 1:100 to minimize the matrix effects to
limited number of excess charges, and to which the target compounds were achieve acceptable recoveries based
the limited space on the surface of the diluted. Dilution typically causes the on a solvent calibration. In the 1:100
charged droplet. The dilution of the final concentrations of the pesticides dilution, all detectable pesticides showed
matrix frees up space at the surface, to increase until the point at which full recovery and basically no signal
resulting in more efficient ionization complete recovery is achieved. Table 3 suppression. This is in agreement with
of the target compounds. In addition, shows the recoveries in black tea extract published results, which showed that the
the amount of matrix injected to the for 10 pesticides and different dilution Agilent Jet Stream Ionization required
LC/MS system is limited, which results ratios. less dilution to eliminate matrix effects
in increased robustness of the analytical compared to equivalent techniques3.
method, minimization of instrument
contamination, and increased instrument
uptime.

6
 10 ng/g in black tea 10 ng/g in black tea, 1–5 dil 10 ng/g in black tea, 1–10 dil 10 ng/g in black tea, 1–20 dil 10 ng/g in black tea, 1–50 dil 10 ng/g in black tea,
[Alanycarb] [Alanycarb] [Alanycarb] [Alanycarb] [Alanycarb] 1–100 dil [Alanycarb]
400.1 & 238.0 Area = 17,039 400.1 & 238.0 Area = 23,661 400.1 & 238.0 Area = 14,572 400.1 & 238.0 Area = 5,812 400.1 & 238.0 Area = 2,011 400.1 & 238.0 Area = 1,294
400.1 & 91.0 Area = 4,165 ×10 3
×10 3 400.1 & 91.0 Area = 4,817 ×10 3 400.1 & 91.0 Area = 7,722 ×10 3 400.1 & 91.0 Area = 2,064
×10 3
400.1 & 91.0 Area = 833
×10 3
400.1 & 91.0 Area = 219
3.4 3.4 1:5 3.4 1:10 3.4 1:20 3.4 1:50 3.4 1:100
3.0 3.0 3.0 3.0 3.0 3.0
2.6 2.6 2.6 2.6 2.6 2.6
2.2 2.2 2.2 2.2 2.2 2.2
1.8 1.8 1.8 1.8 1.8 1.8
1.4 1.4 1.4 1.4 1.4 1.4
1.0 1.0 1.0 1.0 1.0 1.0
0.6 0.6 0.6 0.6 0.6 0.6
0.2 0.2 0.2 0.2 0.2 0.2
11.6 11.8 12.0 11.6 11.8 12.0 11.6 11.8 12.0 11.6 11.8 12.0 11.6 11.8 12.0 11.6 11.8 12.0

10 ng/g in black tea 10 ng/g in black tea, 1–5 dil 10 ng/g in black tea, 1–10 dil 10 ng/g in black tea, 1–20 dil 10 ng/g in black tea, 1–50 dil 10 ng/g in black tea,
[Oxamyl] [Oxamyl] [Oxamyl] [Oxamyl] [Oxamyl] 1–100 dil [Oxamyl]
237.0 & 72.1 Area = 45,373 237.0 & 72.1 Area = 22,084 237.0 & 72.1 Area = 13,029 237.0 & 72.1 Area = 8,135 237.0 & 72.1 Area = 3,354 237.0 & 72.1 Area = 1,741
×10 3 237.0 & 90.1 Area = 7,955 ×10 3 237.0 & 90.1 Area = 3,906 ×10 3 237.0 & 90.1 Area = 2,329 ×10 3 237.0 & 90.1 Area = 1,436 ×10 3 237.0 & 90.1 Area = 604 ×10 3 237.0 & 90.1 Area = 336
8 8 1:5 8 1:10 8 1:20 8 1:50 8 1:100
7 7 7 7 7 7
6 6 6 6 6 6
5 5 5 5 5 5
4 4 4 4 4 4
3 3 3 3 3 3
2 2 2 2 2 2
1 1 1 1 1 1

3.5 3.7 3.9 4.1 3.5 3.7 3.9 4.1 3.5 3.7 3.9 4.1 3.5 3.7 3.9 4.1 3.5 3.7 3.9 4.1 3.5 3.7 3.9 4.1

Figure 4. Comparison of peak areas for alanycarb and oxamyl spiked in black tea and diluted with acetonitrile 1:5, 1:10, 1:20, 1:50, and 1:100 prior to injection.

Table 3. Recoveries for selected pesticides calculated for different dilution ratios. Cells shaded in green comply with requirements of SANCO/12571/2013.

Analytes No dilution (n = 5) Dilution 1:5 (n = 5) Dilution 1:10 (n = 5) Dilution 1:20 (n = 5) Dilution 1:50 (n = 5) Dilution 1:100 (n = 5)
Acetamiprid 29.4 ± 0.8 57.3 ± 1.4 67.5 ± 3.7 79.9 ± 2.9 91.8 ± 5.2 109.5 ± 3.4
Alanycarb 10.4 ± 1.3 73.9 ± 2.2 81.5 ± 14.3 85.7 ± 11.1 87.6 ± 4.7 121.7 ± 10.8
Aldicarb 36.9 ± 1.0 69.9 ± 1.4 78.0 ± 3.5 91.0 ± 4.2 95.2 ± 8.8 104.9 ± 14.1
Carbaryl 56.9 ± 1.8 80.1 ± 3.8 80.8 ± 4.1 96.1 ± 7.2 102.6 ± 6.6 116.4 ± 9.6
Dimethoate 33.9 ± 1.7 68.6 ± 2.4 84.1 ± 5.4 89.0 ± 7.9 88.2 ± 8.8 84.7 ± 7.5
Diuron 79.7 ± 4.0 90.4 ± 7.0 91.7 ± 4.9 94.9 ± 7.2 89.2 ± 7.3 100.9 ± 13.5
Flufenoxuron 95.4 ± 1.1 88.8 ± 1.6 89.4 ± 3.8 93.3 ± 5.8 100.0 ± 6.1 119.2 ± 13.9
Monocrotophos 4.6 ± 0.3 13.9 ± 0.3 21.8 ± 0.8 33.8 ± 1.1 58.5 ± 2.0 95.1 ± 5.7
Oxamyl 20.8 ± 0.7 52.6 ± 1.9 65.0 ± 2.0 79.7 ± 3.0 91.2 ± 4.6 110.6 ± 5.2
Thiamethoxam 40.0 ± 1.4 45.9 ± 0.9 46.6 ± 3.8 52.2 ± 1.7 70.9 ± 2.9 97.3 ± 2.0

7
Conclusions References
An UHPLC/MS/MS based multi-residue 1. Regulation (EC) No 396/2005 of
method for the determination of more the European Parliament and of
than 250 pesticides and pesticide the Council of 23 February 2005 on
metabolites has been developed. It maximum residue levels of pesticides
takes full advantage of the low delay in or on food and feed of plant and
volumes of the Agilent 1290 Infinity LC animal origin (including amendments
System and its ability to handle high as of 18 March 2008) and complying
back pressures in UHPLC separations for with regulation (EC) 1107/2009.
increased chromatographic resolution.
The method benefits from the highly 2. Guidance document on analytical
sensitive Agilent 6495 Triple Quadrupole quality control and validation
LC/MS System and from the versatile procedures for pesticide residues
ionization capabilities of the Agilent Jet analysis in food and feed.
Stream ionization source. Dynamic MRM Document N° SANCO/12571/2013.
acquisition and fast polarity switching Implemented by 01/01/2014.
were used to maximize dwell times http://www.eurl-pesticides.eu/
for each individual compound. Source library/docs/allcrl/AqcGuidance_
parameters were optimized to achieve Sanco_2013_12571.pdf
good sensitivity across the suite of target
3. Stahnke, H., et al. Reduction of Matrix
compounds.
Effects in Liquid Chromatography−
The method was applied to the analysis Electrospray Ionization−Mass
of pesticides in complex matrixes such as Spectrometry by Dilution of the
black tea. Enhanced sensitivity allowed Sample Extracts: How Much Dilution
for more flexibility in the degree of sample is Needed? Anal. Chem. 2012, 84,
dilution. With any dilution, a lower matrix pp 1474-1482 (incl. supporting
amount is introduced into the LC/MS information).
system leading not only to fewer matrix
4. Parra, N.P., Taylor, L. Why Instrument
effects, but also to improved method
Detection Limit (IDL) is a Better
robustness and increased instrument
Metric for Determining the Sensitivity
uptime. Extensive dilution of sample
of Triple Quadrupole LC/MS Systems.
extracts was applied to minimize matrix
Agilent Technologies Technical
effects, and to allow quantitation of all
Overview, publication number
pesticides within the acceptable recovery
5991‑4089EN, 2014.
range of 70 to 120 % based on a solvent
calibration. The increased sensitivity
of the 6495 Triple Quadrupole LC/MS
System allowed the quantitation of the
majority of all targeted pesticides below
the maximum residue limits specified www.agilent.com/chem
by the European Commission, even in
the 1:100 diluted extracts with improved This information is subject to change without notice.
precision and excellent robustness.
© Agilent Technologies, Inc., 2014
Published in the USA, June 13, 2014
5991-4687EN