planar chromatography meets direct ambient mass spectrometry: … · 2015-07-30 · planar...
TRANSCRIPT
Planar chromatography meets direct ambient mass spectrometry: current trends
Elizabeth Crawford1,2, Brian Musselman1 1 IonSense, Inc. Saugus, MA, USA 2 Institute of Chemical Technology, Prague, Czech Republic
2-4. July 2014 HPTLC 2014 Lyon, France
Major Hyphenations of Open Air Ambient MS
& (HP)TLC Separations
DART – Direct Analysis in Real Time
FAPA – Flowing Atmospheric Pressue Afterglow
LA-DART – Laser Ablation DART
LAESI – Laser Ablation Electrospray Ionization
DESI – Desorption Electrospray Ionization
EASI – Easy Ambient Sonic-spray Ionization
LESA – Liquid Extraction Surface Analysis
• TLC-CAMAG ESI Interface – main ESI based method
Thermal Desorption
Laser Ablation
Liquid Spray
Focusing on Open Air Thermal Ionization
Coupled with Mass Spectrometry
No use of solvents during ionization Only semi-destructive - all material not completely
ablated → can run repeat analyses or other methods
Complementary MS data compared with HPTLC ESI-MS – no salt adduct formation, potential thermal separation, both RP & NP plates • → Heating Ramp (RT – 550° C)
Silica gel not disturbed from plate surface → interaction only with heated gas → no source contamination
Major benefits compared to liquid based methods:
DART TLC Evolutions...
DART-100 Yrs: 2006-09
DART-100 & DART-SVP Yrs: 2006-present
DART-SVP at Angle Yrs: 2009-present
Laser Ablation DART 2010 1st Publication 2012 1st TLC DART
Direct TLC Analysis: Vertical On-Edge Method
First coupling of DART with TLC: Morlock, G.; Schwack, W. Determination of isopropylthioxanthone (ITX) in milk, yoghurt and fat by HPTLC-FLD, HPTLC-ESI/MS and HPTLC-DART/MS. Anal. Bioanal. Chem. 2006, 385(3): 586-595.
Sample preparation & separation via HPTLC (Methanolic Extract)
(Silica gel 60 F254 at thickness of 200 µm) Helium at 350° C 0,5 mm/s speed
Direct analysis vertical edge of HPTLC plate
E. Crawford et al. ASMS 2014 ThP 413. Samples provided by Prof. Jason Shepard; University at Albany, SUNY, Albany, NY USA
Benzedrine Tablet: MeOH extract separated by HPTLC (silica gel 60 F254 at thickness of 200 µm)
Direct TLC Analysis: Horizontal Method
Optim. by J.P. Yang et al. at Kyung
Hee University
Seoul, South Korea
Data provided by H.J. Kim and Dr. Y.P. Jang (2011) ; College of Pharmacy Kyung Hee University Seoul, South Korea
Normal Phase (NP) Silica Plates (cut narrow)
A small dab of glycerol was added to the surface of the TLC plate to enhance the heating of the target area on the TLC plate. The TLC plate was mechanically
moved under the DART-SVP source at 0.3 mm/s speed.
Direct TLC Analysis:
45° Angle
Chernetsova, E.S.; Revelsky, A.I.; Morlock, G.E. Some new features of Direct Analysis in Real Time mass spectrometry utilizing the desorption at an angle option. Rapid Comm. Mass Spec., 2011, 25(16): 2275-2282.
Direct TLC Plate Analysis Workflow
A: 350° C heater setting (Compound 1) B: 450° C heater setting (Compound 2) C: 450° C plus glycerol on spot (Cmpds 2&3)
Developed TLC plates are
ready for direct mass
spectrometric analysis
A B C
TLC plate is placed into the holder, which is mounted onto a motorized linear rail
TLC Plate: Pharma Compound 3
RT: 5.07 - 11.23
5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0 10.5 11.0
Time (min)
0
10
20
30
40
50
60
70
80
90
100
Re
lative
Ab
un
da
nce
6.50
6.51
5.94 7.895.95 7.906.935.38 9.348.377.92
5.376.96 9.33
7.936.549.318.387.43 8.367.437.425.96 8.888.855.39
7.46 8.90 9.35 9.757.94 8.92
8.83 9.739.775.99 8.35 8.42 9.72
8.937.476.99 9.37
8.43 9.387.009.82
7.02
10.2110.525.42
8.33 10.898.81
NL: 2.84E5
m/z= 833.99-834.19 MS DART_SVP_In-line_Millennium TLC Plate_MPI 3_450C_with Glycerol_20120404
Replicate of 10 Repeat Sampling of the Same TLC Spot: Glycerol reapplied to single TLC spot before reintroducing the spot to the DART source
Replicates n = 3: NO Glycerol applied
450⁰ C
SS1: Well Separated - Lower & Upper Spots SS1 Plate_Lower_Upper Spots_01 8/3/2012 3:14:24 PM SS1 Plate
He; 350C without/with Glycerol, Full MS
RT: 0.00 - 3.47
0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 3.4
Time (min)
0
20
40
60
80
100
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
1.431.41
1.40
1.39 1.44
1.451.38 1.48
1.361.34 1.501.33
1.521.302.60 3.01 3.352.70 2.84 3.09 3.220.250.15 1.571.25 2.550.300.10 2.23 2.382.141.70 2.052.001.820.61 0.650.52 0.910.81 1.060.47
2.42 2.442.382.48
2.56
2.352.342.56
2.332.57
2.31
2.292.27
2.61 2.63 2.78 3.02 3.09 3.282.95 3.393.230.310.15 2.250.650.10 2.050.76 1.440.61 1.971.521.39 1.71 1.850.51 1.110.81 0.93 1.631.26
NL:7.15E6
m/z= 233.29-237.68 MS SS1 Plate_Lower_Upper Spots_01
NL:2.42E7
m/z= 334.27-339.82 MS SS1 Plate_Lower_Upper Spots_01
200 250 300 350 400 450 500 550 600 650 700
m/z
0
20
40
60
80
100
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
234.17
206.17 311.75 466.25240.17 288.17266.25 403.92391.17346.83 623.92449.75417.67 511.83 536.00487.67 547.83 610.25 685.08587.50 651.75
337.17
670.25212.17 257.25 414.67234.17 435.83351.08291.25 309.17 368.08 449.67396.75 504.67 624.42590.50468.42 639.58556.42536.25 684.33
NL: 5.35E5
SS1 Plate_Lower_Upper Spots_01#510-567 RT: 1.35-1.49 AV: 58 T: ITMS + p NSI Full ms [80.00-1000.00]
NL: 1.05E6
SS1 Plate_Lower_Upper Spots_01#886-994 RT: 2.31-2.56 AV: 109 T: ITMS + p NSI Full ms [80.00-1000.00]
Positive Ion Mode Lower Spot
m/z 234
Positive Ion Mode Upper Spot
m/z 335
Lower Spot
Upper Spot
For the Next Generations:
Laser Ablation TLC Plasma (DART) Ionization
Zhang, J.; Zhou, Z.; Yang, J.; Zhang, W.; Bai, Y.; Liu, H. Thin Layer Chromatography/Plasma Assisted Multiwavelength Laser Desorption Ionization Mass Spectrometry for Facile Separation and Selective Identification of Low Molecular Weight Compounds. Anal. Chem., 2012, 84(3): 1496–1503.
Abstract: “The PAMLDI-MS system was successfully applied in the detection of low molecular weight compounds from different kinds of samples separated on a normal-phase silica gel, such as dye mixtures, drug standards, and tea extract, with the detection level of 5 ng/mm2.”
Conclusions
Both RP & NP plates (glass backed) can be directly analyzed without source contamination
Plates can be cut thin (5 mm wide) and run with DART source in-line with MS
To analyze intact large format plates, set the DART-SVP source at an angle (45° angle) to the plate
Resolution of classic DART-SVP gas beam 3-4 mm, with laser target area becomes 150-400 µm spot size
Coupling with high resolution accurate mass (HRAM) MS is major trend with coupling TLC and ambient MS
Direct quantitation of HPTLC plates with DART-MS → Tim Häbe & Prof. G. Morlock • O15 lecture “Quantitative HPTLC surface analysis by DART-MS
scanning”
DART (HP)TLC Publications
2006 Morlock, G.; Schwack, W. Determination of isopropylthioxanthone (ITX) in milk, yoghurt and fat by HPTLC-FLD, HPTLC-ESI/MS and HPTLC-DART/MS. Anal. Bioanal. Chem., 2006, 385(3): 586-595. 2007 Morlock, G.; Ueda, Y. J. New coupling of planar chromatography with direct analysis in real time mass spectrometry. Chromatogr. A, 2007, 1143: 243−251. 2008 Dytkiewitz, E. and Morlock, G.E. Analytical strategy for rapid identification and quantification of lubricant additives in mineral oil by high-performance thin-layer chromatography with UV absorption and fluorescence detection combined with mass spectrometry and infrared spectroscopy. Journal of AOAC International, 2008, 91(5):1237-1243. Smith, N.J.; Domin, M.A.; Scott, L.T. HRMS Directly From TLC Slides. A Powerful Tool for Rapid Analysis of Organic Mixtures. Org. Lett., 2008, 10(16): 3493-3496. 2009 Jee, E. H.; Jeong, C. W.; Jeong, S. D.; Kim, H. J.; Jang, Y. P., Detection of characterising compounds on TLC by DART-MS. Planta Med, 2009, 75(9): 38. 2010 Kim, H. J.; Jee, E. H.; Ahn, K. S.; Choi, H. S.; Jang, Y. P. Identification of marker compounds in herbal drugs on TLC with DART-MS. Archives of Pharmacal Research, 2010, 33(9): 1355-1359. Kim, H. J.; Oh, M. S.; Hong, J.; Jang, Y. P., Quantitative analysis of major dibenzocyclooctane lignans in schisandrae fructus by online TLC-DART-MS. Phytochemical Analysis, 2010, 22(3): 258-262. 2011 Cheng, S.-C.; Huang, M.-Z.; Shiea, J. Thin layer chromatography/mass spectrometry. Journal of Chromatography A, 2011, 1218(19): p. 2700-2711. Chernetsova, E.S.; Revelsky, A.I.; Morlock, G.E. Some new features of Direct Analysis in Real Time mass spectrometry utilizing the desorption at an angle option. Rapid Comm. Mass Spec., 2011, 25(16): 2275-2282. Howlett, S.E.; Steiner, R.R., Validation of Thin Layer Chromatography with AccuTOF-DART™ Detection for Forensic Drug Analysis. Journal of Forensic Sciences, 2011, 56(5): 1261-1267. Kim, H.J.; Oh, M.S.; Hong, J.; Jang, Y.P. Quantitative analysis of major dibenzocyclooctane lignans in schisandrae fructus by online TLC-DART-MS. Phytochemical Analysis, 2011, 22(3): 258-262. Wood, J.L.; Steiner, R.R. Purification of pharmaceutical preparations using thin-layer chromatography to obtain mass spectra with Direct Analysis in Real Time and accurate mass spectrometry. Drug Testing and Analysis, 2011, 3(6): 345-351. 2012 Morlock, G.E.; Chernetsova, E.S. Coupling of planar chromatography with direct analysis in real time mass spectrometry (DART-MS). Cent Eur J. Chem. 2012, 10(3): 703-710. Zhang, J.; Zhou, Z.; Yang, J.; Zhang, W.; Bai, Y.; Liu, H. Thin Layer Chromatography/Plasma Assisted Multiwavelength Laser Desorption Ionization Mass Spectrometry for Facile Separation and Selective Identification of Low Molecular Weight Compounds. Anal. Chem., 2012, 84(3): 1496–1503. 2013 Djelal, H.; Cornée, C.; Tartivel, R.; Lavastre, O.; Abdeltif, A. The use of HPTLC and Direct Analysis in Real Time-Of-Flight Mass Spectrometry (DART-TOF-MS) for rapid analysis of degradation by oxidation and sonication of an azo dye. Arabian Journal of Chemistry, 2013, (0).
