Microdialysis coupled on-line to capillary liquid chromatography with tandem mass spectrometry for monitoring acetylcholine in vivo

Shackman, Holly Merideth; Shou, Minshan; Cellar, Nicholas A.; Watson, Christopher J.; Kennedy, Robert T.

doi:10.1016/j.jneumeth.2006.06.020

Cited by 54 publications

(40 citation statements)

References 23 publications

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“…High-resolution mass spectral analysis of ACh has been applied to either mammals or in vitro enzyme assays previously. 16,17 However, there have been no reports on the detection of ACh from plant tissues using mass spectrometry at high resolution where the level of ACh is generally much lower compared to animal tissues. In this study, we detected various peaks which have m/z values quite similar, but clearly distinct from that of ACh by ESI-orbitrap FT-MS (Fig.…”

Section: Discussionmentioning

confidence: 99%

High-resolution mass spectrometry for detecting Acetylcholine in Arabidopsis

Murata

Watanabe

Sugahara

et al. 2015

Plant Signaling & Behavior

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Acetylcholine (ACh) was first identified a century ago, and has long been known as a neurotransmitter in animals. However, it has been shown recently that the occurrence of ACh is widespread among various non-animal species including higher plants. Although previous reports suggest that various plant species are capable of responding to exogenously applied ACh, the molecular basis for ACh biosynthesis and regulatory mechanisms mediated by endogenous ACh are largely unclear. This is partly because of the lack of conclusive data on the occurrence and the tissue specificity of ACh in plants. To this end, we performed various analyses including liquid chromatography electrochemical detection (LC-ECD), liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis. The results, together with electrospray ionization-orbitrap Fourier transform mass spectrometry (ESI-orbitrap FT-MS) analysis provide strong evidence that ACh exists in Arabidopsis thaliana tissues. The results also showed that the level of ACh is highest in seed, followed by root and cotyledon. Moreover, exogenously applied ACh inhibited the elongation of Arabidopsis root hairs. These results collectively indicate that ACh exists primarily in seed and root in Arabidopsis seedlings, and plays a pivotal role during the initial stages of seedling development by controlling root hair elongation in Arabidopsis.

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Section: Discussionmentioning

confidence: 99%

High-resolution mass spectrometry for detecting Acetylcholine in Arabidopsis

Murata

Watanabe

Sugahara

et al. 2015

Plant Signaling & Behavior

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“…13 C-lactate, 13 C-pyruvate, glutamine-d 5 , and glutamate-d 5 were used as their respective internal standards (IS). Stock solutions of the analytes were prepared in 30% acetonitrile containing 0.1% formic acid and stored at 5°C.…”

Section: Preparation Of Calibration Standards and Internal Standardsmentioning

confidence: 99%

“…MS scans were split into three segments considering the different MS behaviors of five compounds. In the first segment (0.0-3.5 min) acetylcholine and acetylcholine-d 9 were monitored under a positive mode with the capillary voltage of 3,500 V, while lactate, 13 C-lactate, pyruvate and 13 C-pyruvate were monitored under a negative mode in the second segment (3.5-8.6 min) with the capillary voltage of 2,541 V, and glutamine, glutamine-d 5 , glutamate, and glutamate-d 5 were monitored under a positive mode in the third segment (8.6-15.0 min) with capillary voltage of 3200 V. The MS parameters of five analytes and their corresponding internal standards are listed in Table 1. Lactate and pyruvate did not generate significant and stable product ions, so we could not use the product ions for lactate and pyruvate in MRM conditions.…”

Section: Microdialysis Samplingmentioning

confidence: 99%

“…Recent analytical advances have been made by using MS detection coupled with ion-pair reversed-phase liquid chromatography (RPLC) or ion exchange chromatography (IEC) as separation tools. For instance, acetylcholine has been analyzed by strong cation exchange (SCX)-MS [13] or ion-pair RPLC-MS [14,15]. Pyruvate was analyzed by weak anion exchange (WAX)-MS [16], while lactate and pyruvate together were detected by RPLC-ESI/ MS (electro spray ionization, ESI), but a time-consuming derivatization process with 3-nitrophenyl-hydrazine was needed [17].…”

Section: Introductionmentioning

confidence: 99%

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Measuring multiple neurochemicals and related metabolites in blood and brain of the rhesus monkey by using dual microdialysis sampling and capillary hydrophilic interaction chromatography–mass spectrometry

Pföstl

Zaldívar

et al. 2011

Anal Bioanal Chem

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In vivo measurement of multiple functionally related neurochemicals and metabolites (NMs) is highly interesting but remains challenging in the field of basic neuroscience and clinical research. We present here an analytical method for determining five functionally and metabolically related polar substances, including acetylcholine (quaternary ammonium), lactate and pyruvate (organic acids), as well as glutamine and glutamate (amino acids). These NMs are acquired from samples of the brain and the blood of non-human primates in parallel by dual microdialysis, and subsequently analyzed by a direct capillary hydrophilic interaction chromatography (HILIC)-mass spectrometry (MS) based method. To obtain high sensitivity in electrospray ionization (ESI)-MS, lactate and pyruvate were detected in negative ionization mode whereas the other NMs were detected in positive ionization mode during each HILIC-MS run. The method was validated for linearity, the limits of detection and quantification, precision, accuracy, stability and matrix effect. The detection limit of acetylcholine, lactate, pyruvate, glutamine, and glutamate was 150 pM, 3 μM, 2 μM, 5 nM, and 50 nM, respectively. This allowed us to quantitatively and simultaneously measure the concentrations of all the substances from the acquired dialysates. The concentration ratios of both lactate/pyruvate and glutamine/glutamate were found to be higher in the brain compared to blood (p<0.05). The reliable and simultaneous quantification of these five NMs from brain and blood samples allows us to investigate their relative distribution in the brain and blood, and most importantly paves the way for future non-invasive studies of the functional and metabolic relation of these substances to each other.

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“…Much work has been published on detecting and quantitating targeted quats in differing matrices [14][15][16][17][18][19]. Detection methods that require ion formation, such as mass spectrometry (MS), take advantage of the permanent positive charge already present on quaternary ammonium cations.…”

Section: Introductionmentioning

confidence: 99%

A Novel Route to Recognizing Quaternary Ammonium Cations Using Electrospray Mass Spectrometry

Shackman

Ding

Bolgar

2014

J. Am. Soc. Mass Spectrom.

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Abstract. Characterizing and elucidating structures is a commonplace and necessary activity in the pharmaceutical industry with mass spectrometry and NMR being the primary tools for analysis. Although many functional groups are readily identifiable, quaternary ammonium cations have proven to be difficult to unequivocally identify using these techniques. Due to the lack of an N-H bond, quaternary ammonium groups can only be detected in the 1 H NMR spectra by weak signals generated from long-range 14 N-H coupling, which by themselves are inconclusive evidence of a quaternary ammonium functional group. Due to their low intensity, these signals are frequently not detected. Additionally, ions cannot be differentiated in a mass spectrum as an M + or [M + H] + ion without prior knowledge of the compound's structure. In order to utilize mass spectrometry as a tool for determining this functionality, ion cluster formation of quaternary ammonium cations and non-quaternary amines was studied using electrospray ionization. Several mobile phase modifiers were compared; however, the addition of small amounts of trifluoroacetic acid proved superior in producing characteristic and intense [M +2TFA] -clusters for compounds containing quaternary ammonium cations when using negative electrospray. By fragmenting this characteristic ion using CID, nearly all compounds studied could be unambiguously identified as containing a quaternary ammonium cation or a non-quaternary amine attributable to the presence (non-quaternary amine) or absence (quaternary ammonium cation) of the resulting [2TFA + H] -ion in the product spectra. This method of analysis provides a rapid, novel, and reliable technique for indicating the presence of quaternary ammonium cations in order to aid in structural elucidation.

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Microdialysis coupled on-line to capillary liquid chromatography with tandem mass spectrometry for monitoring acetylcholine in vivo

Cited by 54 publications

References 23 publications

High-resolution mass spectrometry for detecting Acetylcholine in Arabidopsis

High-resolution mass spectrometry for detecting Acetylcholine in Arabidopsis

Measuring multiple neurochemicals and related metabolites in blood and brain of the rhesus monkey by using dual microdialysis sampling and capillary hydrophilic interaction chromatography–mass spectrometry

A Novel Route to Recognizing Quaternary Ammonium Cations Using Electrospray Mass Spectrometry

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