Planar chromatography meets direct ambient mass spectrometry: current trends
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1 Planar chromatography meets direct ambient mass spectrometry: current trends Elizabeth Crawford 1,2, Brian Musselman 1 1 IonSense, Inc. Saugus, MA, USA 2 Institute of Chemical Technology, Prague, Czech Republic 2-4. July 2014 HPTLC 2014 Lyon, France
2 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 Thermal Desorption Laser Ablation Liquid Spray TLC-CAMAG ESI Interface main ESI based method
3 Focusing on Open Air Thermal Ionization Coupled with Mass Spectrometry Major benefits compared to liquid based methods: 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
4 DART TLC Evolutions... DART-100 & DART-SVP Yrs: 2006-present DART-100 Yrs: Laser Ablation DART st Publication st TLC DART DART-SVP at Angle Yrs: 2009-present
5 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): Direct TLC Analysis: Vertical On-Edge Method Sample preparation & separation via HPTLC (Methanolic Extract) (Silica gel 60 F 254 at thickness of 200 µm) Helium at 350 C 0,5 mm/s speed
6 Direct analysis vertical edge of HPTLC plate Benzedrine Tablet: MeOH extract separated by HPTLC (silica gel 60 F 254 at thickness of 200 µm) E. Crawford et al. ASMS 2014 ThP 413. Samples provided by Prof. Jason Shepard; University at Albany, SUNY, Albany, NY USA
7 Direct TLC Analysis: Horizontal Method Optim. by J.P. Yang et al. at Kyung Hee University Seoul, South Korea
8 Normal Phase (NP) Silica Plates (cut narrow) Data provided by H.J. Kim and Dr. Y.P. Jang (2011) ; College of Pharmacy Kyung Hee University Seoul, South Korea
9 Direct TLC Analysis: 45 Angle 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. 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):
10 Direct TLC Plate Analysis Workflow Developed TLC plates are ready for direct mass spectrometric analysis TLC plate is placed into the holder, which is mounted onto a motorized linear rail A: 350 C heater setting (Compound 1) B: 450 C heater setting (Compound 2) C: 450 C plus glycerol on spot (Cmpds 2&3) A B C
11 TLC Plate: Pharma Compound 3 Replicate of 10 Repeat Sampling of the Same TLC Spot: Glycerol reapplied to single TLC spot before reintroducing the spot to the DART source 450⁰ C RT: NL: 2.84E5 m/z= MS DART_SVP_Inline_Millennium TLC Plate_MPI 3_450C_with Glycerol_ Replicates n = 3: NO Glycerol applied Time (min) 8.81
12 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: Positive Ion Mode Lower Spot m/z Positive Ion Mode Upper Spot m/z Time (min) NL: 7.15E6 m/z= MS SS1 Plate_Lower_U pper Spots_01 NL: 2.42E7 m/z= MS SS1 Plate_Lower_U pper Spots_ Lower Spot NL: 5.35E5 SS1 Plate_Lower_Upper Spots_01# RT: AV: 58 T: ITMS + p NSI Full ms [ ] Upper Spot NL: 1.05E6 SS1 Plate_Lower_Upper Spots_01# RT: AV: 109 T: ITMS + p NSI Full ms [ ] m/z
13 For the Next Generations: Laser Ablation TLC Plasma (DART) Ionization 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/mm 2. 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):
14 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 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
15 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): Morlock, G.; Ueda, Y. J. New coupling of planar chromatography with direct analysis in real time mass spectrometry. Chromatogr. A, 2007, 1143: 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): 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): 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): 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): 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): Cheng, S.-C.; Huang, M.-Z.; Shiea, J. Thin layer chromatography/mass spectrometry. Journal of Chromatography A, 2011, 1218(19): p 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): 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): 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): 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): 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): 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): 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).
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