LTQ-Tune mm mm Agilent Technologies ROE 2. 2 ieesi-ms 1 2 mm 5 ~ μl /min 5. 5 kv 1 ieesi-ms Fig. 1 Schematic diagram of inte

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1 檵檵檵檵檵檵檵檵檵檵檵檵檵檵檵檵 FENXI HUAXUE Chinese Journal of Analytical Chemistry ~ 334 檵檵 DOI / j. issn * CH 3 OH /H 2 O CH 3 OH-H 2 O V /V ieesi-ms 1 min DESI 7 8 LAE- SI 9 ieesi-ms 10 ~ 12 ieesi-ms 21 CH 3 OH-H 2 O CH 3 OH-H 2 O V /V ieesi LTQ-XL Xcalibur Thermo Scientific m /z 50 ~ kv V 100 V 10 ~ 30% 30 ms 1. 5 Da Nos No. IRT13054 * chw8868@ gmail. com

160034 1 2 1 2 * 1 1 330013 2 330006 21 CH 3 OH /H 2 O CH 3 OH-H 2 O 30 70 V /V ieesi-ms 1 min 1 1 2 3 4 5 6 DESI 7 8 LAE- SI 9

2 LTQ-Tune mm mm Agilent Technologies ROE 2. 2 ieesi-ms 1 2 mm 5 ~ μl /min 5. 5 kv 1 ieesi-ms Fig. 1 Schematic diagram of internal extractive electrospray 5 cm ionization ieesi mass spectrometry!80 ieesi-ms ieesi-ms CH 3 OH-H 2 O Folch or bligh and dyer recipes 4 CHCl 3 -CH 3 OH 2 1 V /V CH 2 Cl 2 -CH 3 OH 15 - MTBE-CH 3 OH 1 16 CH 3 OH-H 2 O 0 ~ 100% CH 3 OH CA 2a CAB 2b m /z 757 DPPC m /z 783 POPC + + m /z 809 DOPC m /z 833 SAPC a CH 3 OH 0 ~ 15% 4 CH 3 OH CH 3 OH 15% ~ 30% 4 CH 3 OH CH 3 OH 30% ~ 40% CH 3 OH CH 3 OH 35% ~ 75% 4 CH 3 OH CH 3 OH 80% 4 CH 3 OH 30% 4 CH 3 OH 80% 2b CH 3 OH 0 ~ 30% CH 3 OH m /z 757 DPPC + + m /z 783 POPC + + m /z 809 DOPC + m /z 833 SAPC CH 3 OH 35% +

1 Folch or bligh and dyer recipes 4 CHCl 3 -CH 3 OH 2 1 V /V 13 14 - CH 2 Cl 2 -CH 3 OH 15 - MTBE-CH 3 OH 1 16 CH 3 OH-H 2 O 0 ~ 100% 2.

3 CA a CAB b 1 m /z 757 DPPC m /z 783 POPC m /z 809 DOPC m /z 833 SAPC + + CH 3 OH Fig. 2 Effect of solvent constitution of ieesi. 1 dipalmitoyl phosphatidylcholine m /z 757 DPPC phosphatidylcholine m /z 783 POPC oleoyl phosphatidylcholine m /z 809 DOPC + + and 4 arachidonic acid stearoyl phosphatidylcholine m /z 833 SAPC + + from lung cancer tissue a and adjacent tissue b with different proportion of methanol 1. m /z m /z m /z m /z CH 3 OH 40% ~ 70% 4 CH 3 OH CH 3 OH 75% 4 CH 3 OH 30% 4 CH 3 OH 75% CH 3 OH-H 2 O V /V ieesi ieesi 3. 2 CH 3 OH H 2 O V /V 3a 3b 3c ieesi-ms CID 3 m /z m /z 757 PC m /z 773 PC K + m /z 783 PC m /z 799 PC K + m /z 809 PC m /z 825 PC K + m /z 833 PC a m /z 783 PC m /z 799 PC K + m /z 809 PC m /z 825 PC K + m /z 833 PC m /z 757 PC m /z PS 17 PS 2 AT2 DPPC 80% AT2 18 D SP-D 19 B pro-sp-b 20 PS AT2 3a 3c m /z 700 ~ 900 m /z

SAPC + + from lung cancer tissue a and adjacent tissue b with different proportion of methanol 1. m /z 757 2. m /z 783 3. m /z 809 4. m /z 833.

4 CH 3 OH H 2 O V /V ieesi-ms a b c Fig. 3 ieesi-ms analysis of different cancer tissues using CH 3 OH-H 2 O V /V as solvent a lung cancer tissue b lung cancer adjacent tissue c esophageal cancer tissue CH 3 OH-H 2 O V /V 4a 4b 4c 4d ieesi-ms 4 m /z 700 ~ 900 m /z CH 3 OH-H 2 O V /V 4 ieesi-ms 21 CH 3 OH /H 2 O CH 3 OH-H 2 O V /V ieesi-ms

tissue b lung cancer adjacent tissue c esophageal cancer tissue 1 2 5 21 22 3.

5 3 333 References 4 CH 3 OH-H 2 O V /V ieesi-ms a b c d Fig. 4 ieesi-ms analysis of different meat using CH 3 OH-H 2 O V /V as solvent a pork b beef c porcine heart d porcine lung 1 Wang H Manicke N E Yang Q Zheng L Shi R Cooks R G Ouyang Z. Anal. Chem Santos C R Schulze A. FEBS J Ellis S R Ferris C J Gilmore K J Mitchell T W Blanksby S J In Het Panhuis M. Anal. Chem Xie W Gao D Jin F Jiang Y Liu H. Anal. Chem Guo S Wang Y Zhou D Li Z. Sci. Rep Paglia G Ifa D R Wu C Corso G Cooks R G. Anal. Chem Girod M Shi Y Cheng J X Cooks R G. Anal. Chem Lostun D Perez C J Licence P Barrett D A Ifa D R. Anal. Chem Nemes P Woods A S Vertes A. Anal. Chem Zhang H Gu H Yan F Wang N Wei Y Xu J Chen H. Sci. Rep Zhang H Chingin K Zhu L Chen H. Anal. Chem Zhang H Zhu L Luo L Wang N Chingin K Guo X Chen H. J. Agric. Food Chem Nagano K Kano H Arito H Yamamoto S Matsushima T. J. Toxicol. Env. Heal. A XI Cong CHEN Yan-Hua YANG Wei ZHANG Rui-Ping HE Jiu-Ming ZHANG Hua-Bing XIAO Xin-Hua ZEPER Abliz. Chinese J. Anal. Chem

2011 83 4 1197-1201 2 Santos C R Schulze A. FEBS J. 2012 279 15 2610-2623 3 Ellis S R Ferris C J Gilmore K J Mitchell T W Blanksby S J In Het Panhuis M. Anal. Chem.

6 Hu C van Dommelen J van Der Heijden R Spijksma G Reijmers T H Wang M Slee E Lu X Xu G van Der Greef J. J. Proteome Res JIA Bin ZHANG Xing-Lei DING Jian-Hua YANG Shui-Ping CHEN Huan-Wen. Chinese Science Bulletin Chinese Version Wei Y Chen L Zhou W Chingin K Ouyang Y Zhu T Wen H Ding J Xu J Chen H. Sci. Rep Sutherland K D Berns A. Mol. Oncol Ishii T Hagiwara K Ikeda S Arai T Mieno M N Kumasaka T Muramatsu M Sawabe M Gemma A Kida K. COPD Sin D D Tammemagi C M Lam S Barnett M J Duan X Tam A Auman H Feng Z Goodman G E Hanash S. J. Clin. Oncol Zwaal R F A Comfurius P Bevers E M. Cell Mol. Life Sci Aboagye E O Bhujwalla Z M. Cancer Res Direct Analysis of Phospholipids in Biological Tissues Using Internal Exlractive Electrospray Ionization Mass Spectrometry LU Hai-Yan 1 ZHANG Jian-Yong 2 ZHOU Wei 1 WEI Yi-Ping 2 CHEN Huan-Wen * 1 1 East China Institute of Technology Jiangxi Key Laboratory for Mass Spectrometry and Instrumentation Nanchang China 2 Second Affiliated Hospital of Nanchang University Nanchang China Abstract Phospholipids and their metabolites play an important role in a variety of cellular processes including cell-cell adhesion cell growth and differentiation apoptosis phagocytosis as well as storage of energy. In this study the phospholipid composition of cancer tissue and adjacent normal tissue from humans and animals were analyzed by internal extractive electrospray ionization mass spectrometry ieesi-ms. Extractive solvent at high voltage kv was injected into tissue samples using a fused silica capillary at a flow rate of μl /min producing fine charged droplets containing analytes of tissue samples at the tip of the sample. Charged droplets were directly sampled to the atmospheric inlet of a mass spectrometer. Out of 21 different ratios of CH 3 OH H 2 O solvent mixture the ratio CH 3 OH H 2 O = V /V showed the optimal phospholipids extraction and visibility in MS. A large number of phospholipids from different tissue samples such as cancer tissue and adjacent normal tissue of lung cancer esophageal cancer tissue pork beef porcine heart and porcine lung were obtained simultaneously by ieesi-ms analysis. The experimental results demonstrated that ieesi-ms was characterized by minimal sample pretreatment low sample consumption and rapid analysis the analysis time per sample was less than 1 min and the selectivity and sensitivity of ieesi-ms could be improved by choosing proper solvent. Importantly the experimental results provided new information for further studies of phospholipids in biological tissues. Keywords Biological tissue Phospholipids Extractive solvent Internal extractive electrospary inozation mass spectrometry Received 14 January 2016 accepted 20 January 2016 This work was supported by the National Natural Science Foundation of China No No Program for Changjiang Scholars and Innovation Research Team in Universities PCSIRT No. IRT13054

2012 57 20 1918 17 Wei Y Chen L Zhou W Chingin K Ouyang Y Zhu T Wen H Ding J Xu J Chen H. Sci. Rep. 2015 5 10077 18 Sutherland K D Berns A. Mol. Oncol.

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