Strategies for quantitation of endogenous adenine nucleotides in human plasma using novel ion-pair hydrophilic interaction chromatography coupled with tandem mass spectrometry

Zhang, Guodong; Walker, Annie D.; Lin, Zhigui; Han, Xiaogang; Blatnik, Matthew; Steenwyk, Rick C.; Groeber, Elizabeth

doi:10.1016/j.chroma.2013.12.017

Cited by 40 publications

(28 citation statements)

References 41 publications

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“…The LODs and limits of quantitation (LOQ) of the CE-ESI-MS method for many tested analytes were in the low nanomolar range, similar to, or better than, prior reports. 26,27 This performance has lower LODs compared with other nucleotide characterization approaches such as LC-MS (in μg/ml) and HILIC-MS (in ng/ml) 45,46 and is well suited for small-volume samples such as cells.…”

Section: Resultsmentioning

confidence: 99%

Analysis of endogenous nucleotides by single cell capillary electrophoresis-mass spectrometry

Liu

Aerts

Rubakhin

et al. 2014

Analyst

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Analytical technologies that enable investigations at the single cell level facilitate a range of studies; here a lab-fabricated capillary electrophoresis-electrospray ionization-mass spectrometry (CE-ESI-MS) platform was used to analyze anionic metabolites from individual Aplysia californica neurons. The system employs a customized coaxial sheath-flow nanospray interface connected to a separation capillary, with the sheath liquid and separation buffer optimized to ensure a stable spray. The method provided good repeatability of separation and reliable detection sensitivity for 16 mono-, di- and triphosphate nucleosides. For a range of anionic analytes, including cyclic adenosine monophosphate (cAMP), adenosine diphosphate (ADP) and adenosine triphosphate (ATP), the detection limits were in the low nanomolar range (<22 nM). A large Aplysia R2 neuron was used to demonstrate the ability of CE-ESI-MS to quantitatively characterize anionic metabolites within individual cells, with 15 nucleotides and derivatives detected. Following the method validation process, we probed smaller, 60-μm diameter Aplysia sensory neurons where sample stacking was used as a simple on-line analyte preconcentration approach. The calculated energy balance ([ATP] + 0.5 × [ADP])/([AMP] + [ADP] + [ATP]) of these cells was comparable with the value obtained from bulk samples.

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Section: Resultsmentioning

confidence: 99%

Analysis of endogenous nucleotides by single cell capillary electrophoresis-mass spectrometry

Liu

Aerts

Rubakhin

et al. 2014

Analyst

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“…The 400 μl upper aqueous layer was centrifugally filtered through a Millipore cutoff filter to remove the protein content. The filtrate was then centrifugally concentrated and re-suspended in Milli-Q water for the metabolomic analysis at APT via hydrophilic interaction liquid chromatography coupled with tandem mass-spectrometry (HILIC-MS/MS) using previously described methods [20, 21]. …”

Section: Methodsmentioning

confidence: 99%

Maternal low thyroxin levels are associated with adverse pregnancy outcomes in a Chinese population

Zhang

Dai

et al. 2017

PLoS ONE

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Although thyroid dysfunction in early pregnancy may have adverse effects on pregnancy outcomes, few studies have examined the relationship between maternal low free thyroxin (FT4) levels in both first and third trimesters of pregnancy and the incidence of adverse pregnancy outcomes. We hypothesized that low FT4 levels in either first or third trimesters of pregnancy may have different effects on pregnancy outcomes. The study included 6,031 mothers who provided both first and third pregnancy serum samples for analyses of thyroid function. Adverse pregnancy outcomes, such as gestational diabetes mellitus (GDM), pregnancy-induced hypertension and preeclampsia, were diagnosed using the oral glucose tolerance test, blood pressure and urine protein test. Serum metabolites like adenosine and its analogues were identified using hydrophilic interaction liquid chromatography (HILIC)-tandem mass spectrometry (MS/MS). The incidence of hypothyroidism in pregnant women tended to increase with age and pre-pregnancy body mass index (BMI). The incidence of GDM was negatively correlated with maternal FT4 levels during early pregnancy while the incidence of preeclampsia was negatively correlated with maternal FT4 levels during late pregnancy. The incidence of pregnancy-induced hypertension was not significantly correlated with maternal FT4 levels. The women who had isolated maternal hypothyroxemia (IMH) in the third trimester of pregnancy had an increased risk of developing preeclampsia. Some metabolites like adenosine and its analogues in the serum were significantly changed in pregnant mothers with IMH. In conclusion, low FT4 levels during pregnancy are a risk factor for GDM and preeclampsia. Adenosine and its analogues may be important bridges between IMH and preeclampsia.

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“…More recently developed ion‐pair reversed‐phase LC–MS methods yielded LODs in the range from 1 to 10 nM for AMP, ADP, and ATP using multiple reaction monitoring and an injection volume of 5 µL . HILIC–MS‐based approaches recently developed for nucleotide profiling in various biological samples provided LODs typically in the range from 2 to 100 nM using MS/MS mode and employing an injection volume of 5 µL . In this context, the proposed sheathless CZE–MS method (used in full scan MS mode only) provided concentration LODs for the studied nucleotides that are comparable to or better than LODs obtained by LC–MS‐based approaches employing multiple reaction monitoring.…”

Section: Resultsmentioning

confidence: 99%

Profiling nucleotides in low numbers of mammalian cells by sheathless CE–MS in positive ion mode: Circumventing corona discharge

et al. 2020

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Negative ion mode nano‐ESI–MS is often considered for the analysis of acidic compounds, including nucleotides. However, under high aqueous separation conditions, corona discharge is frequently observed at emitter tips, which may result in low ion abundances and reduced nano‐ESI needle emitter lifetimes. In this work, we introduce a sheathless CE‐MS method for the highly efficient and sensitive analysis of nucleotides employing ESI in positive ion mode, thereby fully circumventing corona discharge. By using a background electrolyte of 16 mM ammonium acetate (pH 9.7) a mixture of 12 nucleotides, composed of mono‐, di‐, and tri‐phosphates, could be efficiently analyzed with plate numbers per meter above 220 000 and with LODs in the range from 0.06 to 1.3 nM, corresponding to 0.4 to 8.6 attomole, when using an injection volume of about 6.5 nL only. The utility of the method was demonstrated for the profiling of nucleotides in low numbers of mammalian cells using HepG2 cells as a model system. Endogenous nucleotides could be efficiently analyzed in extracts from 50 000 down to 500 HepG2 cells only. Moreover, apart from nucleotides, also some nicotinamide‐adenine dinucleotides and amino acids could be analyzed under these conditions, thereby clearly illustrating the utility of this approach for metabolic profiling of low amounts of biological material.

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Strategies for quantitation of endogenous adenine nucleotides in human plasma using novel ion-pair hydrophilic interaction chromatography coupled with tandem mass spectrometry

Cited by 40 publications

References 41 publications

Analysis of endogenous nucleotides by single cell capillary electrophoresis-mass spectrometry

Analysis of endogenous nucleotides by single cell capillary electrophoresis-mass spectrometry

Maternal low thyroxin levels are associated with adverse pregnancy outcomes in a Chinese population

Profiling nucleotides in low numbers of mammalian cells by sheathless CE–MS in positive ion mode: Circumventing corona discharge

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