Coated Blade Spray – High Resolution Mass Spectrometry: a versatile tool for

sample profiling and screening of controlled substances in complex matrices

German A. Gómez-Ríos1, Robert Cody2, Nathaly Reyes-Garcés1, Frances A. Carroll1, Gary Stidsen1, David S. Bell1
1Restek Corporation, Bellefonte, Pennsylvania, USA; 2JEOL Corporation, Peabody, Massachusetts, USA

Introduction Experimental (cont.) Results (cont.) Results (cont.)

Coated Blade Spray (CBS) is a technology that allows for analyte Diagram 1 Experimental workflow followed for analysis of solid samples Likewise, CBS was capable of differentiating different types of meat previously attained on a triple quadrupole (QqQ) MS, LOD (1-10 ng/mL)
collection and direct-to-MS interface from a single device [1-4]. (left, meat and fish tissue) and liquid samples (right, beer and urine). In samples. As can be seen in Figure 4, the DAPC-KPCA plot showed clear were acceptable for most of the targeted analytes. In addition, accuracy
Essentially, CBS is a coated stainless steel sheet with the shape of a small the case of liquid samples, the devices were preconditioned, prior to the distinction of each meat. By using 60 PC, the LOOCV and SVM lead to a for the validation points range between 75-112% at the three levels
sword which, thanks to its ultra-thin coating, permits rapid enrichment of sampling, following conditions described elsewhere [4]. predictability of 94 and 96%, respectively. under investigation.
small molecules present in complex samples. Further, it allows for
ionization via ESI mechanism once the coated area is wetted with a small Stick the sample with the pointy Insert CBS into liquid sample
Figure 4 DAPC-KPCA plot classification of miscellaneous fish types (left). Figure 6 Calibration curves for EDDP, Citalopram, LSD, Ketamine, Cocaine
amount of solvent (<15µL) and a high-potential is applied to the non- end for 5 minutes (96-well format) for 15 minutes Heat map representing mass spectra attained for each of the meat and Fentanyl. Validation points at 15, 75 and 150 ng/mL.
coated area of the device[5-7] . Herein, we demonstrate how CBS coupled samples (n=12; right).
18 5 6
y = 0.078x + 0.0358
to High Resolution Mass Spectrometry (HRMS) enables rapid profiling of Rinsed coated area with water Rinsed coated area with water 16 R² = 0.9997 4.5 y = 0.0226x - 0.0293
5
y = 0.0235x + 0.112
R² = 0.9986
4 R² = 0.9982

Ratio [Citalopram/Fentanyl-d5]
aqueous and solid matrices. Unlike other ambient-ionization

Ratio [EDDP/Cocaine-d3]
14
3.5

Ratio [LSD/Cocaine-d3]
4
Store CBS in vacuum sealed bag
12

technologies, CBS allows sampling to be performed remotely, cleaning-

3

Store CBS in vacuum sealed bag and transport to the laboratory

10
2.5 3

up the sample and retaining relevant chemical information that facilitates

8 2
and transport to the laboratory 6 1.5
2

its classification via chemometric tools. Furthermore, we explore CBS- Elute analytes from coating by 4
0.5
1
1

HRMS as a tool for screening and quantitation of controlled substances in Elute analytes from coating by applying 1.5 µL of elution solvent
2

0
0 0

applying 1.5 µL of elution solvent

0 50 100 150 200 0 50 100 150 200

biological matrices such as urine. (MeOH:Water, 95:5, 0.1% FA)

0 50 100 150 200
EDDP in urine samples [ng/mL] Citalopram in urine samples [ng/mL] LSD in urine samples [ng/mL]

(MeOH:Water, 95:5, 0.1% FA) 1.6 8 6

Experimental
y = 0.0065x + 0.0417 y = 0.0332x + 0.1602
1.4 R² = 0.998 7 R² = 0.9997

Ionize
Ionize analytes
analytes of
of interest
interest Ionize analytes of interest 5

Ratio [Ketamine/Cocaine-d3]

Ratio [Cocaine/Cocaine-d3]

Ratio [Fentanyl/Fentanyl-d5]
1.2 6
4
1 5

Coated Blade Spray devices with a coating length of 10 mm and coating Collect mass spectral information Collect mass spectral information 0.8 4 3
y = 0.0234x + 0.1068

thickness of 10 µm were used for all the experiments herein described.

0.6 3 R² = 0.9986
2

The coating consisted of 5 µm Hydrophobic-Lipophilic Balance (HLB) Perform qualitative or quantitative In a third experiment, CBS was used to differentiate diverse fish samples. 0.4 2
1
Derive centroid data and submit to
0.2 1

particles immobilized with polyacrylonitrile. Data collection was analysis As can be seen in Figure 4, the DAPC-KPCA plot showed clear distinction 0 0
Mass Mountaineer for statistical
0

of each species. Similar to the meat samples, by using 60 PC, the LOOCV
0 50 100 150 200 0 50 100 150 200 0 50 100 150 200

conducted on an AccuTOF™ Time-of-Flight MS (JEOL, Peabody, MA) classification Ketamine in urine samples [ng/mL] Cocaine in urine samples [ng/mL] Fentanyl in urine samples [ng/mL]

within the range of 50-1000 m/z. CBS devices were adapted to fit into the and SVM lead to a predictability of 94 and 96%, respectively. Certainly,
further improvements are expected by controlling the spatial location
JEOL PaperSpray™ interface and electrospray was generated by adding
1.5 µL on the coated area and applying a positive voltage of 3.75 kV. Results where the blade is inserted onto the fish for analyte collection. Besides, Summary
Spray time was approximately 10 seconds per blade and the data was our work is also focused on studying a much larger cohort for each of
Sample profiling via mass spectrometry requires taking either the sample 1. CBS was used, as a proof-of-concept for profiling of beer, meat and
processed using msAxel LP. Chemometric analyses were performed with the samples and the exploration of other elution/ionization solvents, so
to the lab or the MS to the sample [8, 9]. Herein, we show how CBS can fish samples. In the case of aqueous matrices, 96 samples can be
Mass Mountaineer™ software for mass spectra collected over the m/z to increase the coverage of analytes under scrutiny (i.e. balance
be used to collect and store chemical information from liquid and solid scrutinized at the same time, allowing for total analysis time under 1
range 80-1000. coverage).
samples. In combination with a HRMS and appropriate statistical tools, minute per sample.
chemicals stored on the coating can be used for sample discrimination. 2. Unlike other methods, were the mass spectrometer must be
Figure 5 DAPC-KPCA plot classification of miscellaneous fish types (left). transported to the place of sampling, sample collection with CBS was
Figure 1 Coated Blade Spray installed on an AccuTOF Time-of-Flight MS As can be seen in Figure 2, the discriminant analysis of principal
Heat map representing mass spectra attained for each of the fish performed in one location (Bellefonte, PA) and analyzed at a different
via commercially available JEOL PaperSpray interface; hence, facilitating components (DAPC) in combination with Kernel Principal Component
samples (n=12; right). location (Peabody, MA) without major hassle as the “sample” (sample
rapid transition between diverse ambient ionization technologies (A-C). Analysis (KPCA) allowed for adequate classification of each of the beer
Insert D presents an example of an ion chronogram collected with CBS in brands under evaluation. Further, when using 60 principal components information) was safely transported on a airplane.
urine sample, while insert B displays Taylor cone generated from CBS (PC), the leave-one-out cross validation (LOOCV) and the Support Vector 3. Although, the limits of quantitation attained via CBS-TOF fall behind
device with a voltage of 3.75 kV and a distance of approximately 4 mm Machine (SVM) unequivocally identified each of the samples. those previously attained with QqQ instruments [4], the results
from the MS inlet. showed the potential towards rapid screening with TOF systems.
Figure 2 DAPC-KPCA plot classification of diverse beer brands (left). 4. Since the chemical information of the sample is stored in the coated
B
TICC / Urine_30min_200 / ESI+ / /

D Photo of the 96-well plate with 12-samples of each beer kind. Sampling material, it is invisible to the bare eye and encrypted to the trained
A
8
x10
4.00

3.75
was performed using a 96-CBS arrangement (right). eye; thus, making CBS an ideal tool, not only for profiling, but also
3.50

3.25
from a chain of custody perspective [9].
3.00

2.75

2.50 References
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Voltage
Intensity

2.00
ON 1. Gómez-Ríos, G.A., et al., Angewandte Chemie, 2014, 52, 1403-1407; 2. Gómez-
C 1.75
Finally, we evaluated the coupling of CBS to the JEOL TOF towards the Ríos, G.A., et al., Bioanalysis, 2018, 10(4), 257–271; 3. Reyes-Garcés., et al.,
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semi-quantitative analysis of several drugs of different classes in human Analytical Chemistry, 2018, 90, 302-360; 4. Tascon, M., et al., Analytical Chemistry,
urine samples. Extraction time was set to 15 minutes and data collection
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1.00 2017, 89, 8421–8428; 5. Gómez-Ríos, G.A., et al., Scientific Reports, 2018, 7, 16104;
0.75
was 5 seconds per blade. Potential inter-blade errors were corrected 6. Poole, J., et al., 2017, Environmental Science & Technology, 2017, 51, 12566–
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Voltage Voltage using two isotopic labelled internal standards. As can be seen in Figure 6, 12572; 7. Gómez-Ríos, G.A., et al., Analyst, 2017, 142, 2928-2935; 8. Cody, R., et al.,
OFF OFF Anal. Chim. Acta., 2017, 989, 38-44; 9. Gómez-Ríos, G.A., et al., Trends in Analytical
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96-well plate limits of quantitation in the low ng/mL range were attained for the drugs
Chemistry., 2019, 112, 201-211.
under evaluation. Though the performance can not be compared to that
1.3 1.4 1.5 1.6 1.7 1.8 1.9

Time [min]
collection