title: In situ derivatization-ultrasound-assisted dispersive liquid-liquid microextraction for the determination of neurotransmitters in Parkinson's rat brain microdialysates by ultra high performance liquid chromatography-tandem mass spectrometry.
abstract: Simultaneous monitoring of several neurotransmitters (NTs) linked to Parkinson's disease (PD) has important scientific significance for PD related pathology, pharmacology and drug screening. A new simple, fast and sensitive analytical method, based on in situ derivatization-ultrasound-assisted dispersive liquid-liquid microextraction (in situ DUADLLME) in a single step, has been proposed for the quantitative determination of catecholamines and their biosynthesis precursors and metabolites in rat brain microdialysates. The method involved the rapid injection of the mixture of low toxic bromobenzene (extractant) and acetonitrile (dispersant), which containing commercial Lissamine rhodamine B sulfonyl chloride (LRSC) as derivatization reagent, into the aqueous phase of sample and buffer, and the following in situ DUADLLME procedure. After centrifugation, 50μL of the sedimented phase (bromobenzene) was directly injected for ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) detection in multiple reaction monitoring (MRM) mode. This interesting combination brought the advantages of speediness, simpleness, low matrix effects and high sensitivity in an effective way. Parameters of in situ DUADLLME and UHPLC-MS/MS conditions were all optimized in detail. The optimum conditions of in situ DUADLLME were found to be 30μL of microdialysates, 150μL of acetonitrile containing LRSC, 50μL of bromobenzene and 800μL of NaHCO3-Na2CO3 buffer (pH 10.5) for 3.0min at 37°C. Under the optimized conditions, good linearity was observed with LODs (S/N>3) and LOQs (S/N>10) of LRSC derivatized-NTs in the range of 0.002-0.004 and 0.007-0.015 nmol/L, respectively. It also brought good precision (3.2-12.8%, peak area CVs%), accuracy (94.2-108.6%), recovery (94.5-105.5%) and stability (3.8-8.1%, peak area CVs%) results. Moreover, LRSC derivatization significantly improved chromatographic resolution and MS detection sensitivity of NTs when compared with the reported studies through the introduction of a permanent charged moiety from LRSC into NTs. Taken together, this in situ DUADLLME method was successfully applied for the simultaneous determination of six NTs in biological samples.
authors.affiliation: Shandong Provincial Key Laboratory of Life-Organic Analysis & Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu 273165, Shandong, PR China.
authors.forename: Yongrui
authors.initials: Y
authors.lastname: He
authors.affiliation: Shandong Provincial Key Laboratory of Life-Organic Analysis & Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu 273165, Shandong, PR China. Electronic address: xianenzhao@163.com.
authors.forename: Xian-En
authors.initials: XE
authors.lastname: Zhao
authors.affiliation: Shandong Provincial Key Laboratory of Life-Organic Analysis & Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu 273165, Shandong, PR China.
authors.forename: Shuyun
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Shandong Provincial Key Laboratory of Life-Organic Analysis & Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu 273165, Shandong, PR China.
authors.forename: Na
authors.initials: N
authors.lastname: Wei
authors.affiliation: Qinghai Key Laboratory of Qinghai-Tibet Plateau Biological Resources & Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Science, Xining 810001, Qinghai, PR China.
authors.forename: Jing
authors.initials: J
authors.lastname: Sun
authors.affiliation: Qinghai Key Laboratory of Qinghai-Tibet Plateau Biological Resources & Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Science, Xining 810001, Qinghai, PR China.
authors.forename: Yubi
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: National Center for Mass Spectrometry in Changchun & Key Laboratory for Traditional Chinese Medicine Chemistry and Mass Spectrometry of Jilin Province, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, 5625 Renmin Street, Changchun 130022, PR China.
authors.forename: Shu
authors.initials: S
authors.lastname: Liu
authors.affiliation: National Center for Mass Spectrometry in Changchun & Key Laboratory for Traditional Chinese Medicine Chemistry and Mass Spectrometry of Jilin Province, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, 5625 Renmin Street, Changchun 130022, PR China. Electronic address: liuzq@ciac.ac.cn.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Shandong Provincial Key Laboratory of Life-Organic Analysis & Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu 273165, Shandong, PR China.
authors.forename: Guang
authors.initials: G
authors.lastname: Chen
authors.affiliation: Qinghai Key Laboratory of Qinghai-Tibet Plateau Biological Resources & Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Science, Xining 810001, Qinghai, PR China.
authors.forename: Yourui
authors.initials: Y
authors.lastname: Suo
authors.affiliation: Shandong Provincial Key Laboratory of Life-Organic Analysis & Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu 273165, Shandong, PR China; Qinghai Key Laboratory of Qinghai-Tibet Plateau Biological Resources & Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Science, Xining 810001, Qinghai, PR China. Electronic address: jmyou6304@163.com.
authors.forename: Jinmao
authors.initials: J
authors.lastname: You
mesh_terms: D000097:Acetonitriles / Q000737:chemistry; D000818:Animals; D001923:Brain Chemistry; D001969:Bromobenzenes / Q000737:chemistry; D002021:Buffers; D002395:Catecholamines / Q000032:analysis* / Q000096:biosynthesis / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D055772:Green Chemistry Technology; D057230:Limit of Detection; D059627:Liquid Phase Microextraction / Q000379:methods*; D017551:Microdialysis; D018377:Neurotransmitter Agents / Q000032:analysis* / Q000096:biosynthesis / Q000378:metabolism; D010300:Parkinson Disease / Q000378:metabolism*; D051381:Rats; D012235:Rhodamines / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013997:Time Factors; D014465:Ultrasonics
chemical_list: D000097:Acetonitriles; D001969:Bromobenzenes; D002021:Buffers; D002395:Catecholamines; D018377:Neurotransmitter Agents; D012235:Rhodamines; C027305:Lissamine Rhodamine B sulfonyl chloride; C032036:bromobenzene; C032159:acetonitrile
keywords: In situ derivatization; In vivo microdialysis; Neurotransmitters; Traditional chinese medicine; Ultra high performance liquid chromatography-tandem mass spectrometry; Ultrasound-assisted dispersive liquid–liquid microextraction