Smith-Lemli-Opitz syndrome diagnosed by using time-of-flight secondary-ion mass spectrometry

Abstract We describe a rapid and sensitive method involving time-of-flight secondary-ion mass spectrometry (TOF-SIMS) for specific laboratory diagnosis of the Smith-Lemli-Opitz syndrome, which is characterized by massive (approximately 1000-fold) accumulation of the biosynthetic cholesterol precursor 7-dehydrocholesterol. Minute amounts of blood (1-50 microL) were extracted with n-hexane, and aliquots were analyzed by TOF-SIMS. 7-Dehydrocholesterol and its isomers were detected at 491.3 mass units ([M + 107Ag]+) and cholesterol at 495.3 mass units ([M + 109Ag]+). Quantitation of 7-dehydrocholesterol and cholesterol was achieved after saponification and addition of stigmasterol as internal standard. Whereas 7-dehydrocholesterol and isomeric dehydrocholesterol were not detectable in controls, the patients revealed concentrations ranging between 0.84 and 1.25 mmol/L. Comparison with results obtained by gas chromatography indicated that quantitation by TOF-SIMS yielded the sum of 7-dehydrocholesterol, isomeric dehydrocholesterol II, and sterol III, the latter two also being increased in the patients. Consistent with quantitation by gas chromatography, the cholesterol concentrations in the patients ranged between 1.54 and 2.12 mmol/L (controls: 6.10 +/- 1.37 mmol/L).

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Distribution of extracts including 4,8-dihydroxy-5-methoxy-2-naphthaldehyde in Diospyros kaki analyzed by gas chromatography-mass spectrometry and time-of-flight secondary ion mass spectrometry

Holzforschung ◽

10.1515/hf-2011-0214 ◽

2012 ◽

Vol 66 (6) ◽

pp. 705-709 ◽

Cited By ~ 6

Author(s):

Yasuyuki Matsushita ◽

In-Cheol Jang ◽

Takanori Imai ◽

Ruka Takama ◽

Kaori Saito ◽

...

Keyword(s):

Mass Spectrometry ◽

Gas Chromatography ◽

Secondary Ion Mass Spectrometry ◽

Time Of Flight ◽

Gas Chromatography Mass Spectrometry ◽

Diospyros Kaki ◽

Compound I ◽

Tof Sims ◽

Ion Mass Spectrometry ◽

Secondary Ion

Abstract The distribution of ethyl acetate extracts and 4,8-dihydroxy-5-methoxy-2-naphthaldehyde (compound I), which is a major constituent of the extracts obtained from the blackened heartwood of Diospyros kaki, was analyzed via gas chromatography-mass spectrometry (GC-MS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS). According to GC-MS, the extracts and compound I are high in concentration at the pith and at the edges of the blackened heartwood. ToF-SIMS analysis revealed a peak at a mass-to-charge ratio of (m/z) 218, which is characteristic of the ionic form of compound I. The ToF-SIMS imaging of compound I in the blackened heartwood based on m/z 218 shows that compound I is located in parenchyma cells and their neighboring axial elements.

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Applications of Time-OF-Flight Secondary Ion Mass Spectrometry (TOF-SIMS)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100135149 ◽

1990 ◽

Vol 48 (2) ◽

pp. 308-309

Author(s):

Bruno Schueler ◽

Robert W. Odom

Keyword(s):

Mass Spectrometry ◽

Secondary Ion Mass Spectrometry ◽

Structural Information ◽

Time Of Flight ◽

Biological Tissues ◽

Polymer Surfaces ◽

Tof Sims ◽

Secondary Ions ◽

Ion Mass Spectrometry ◽

Secondary Ion

Time-of-flight secondary ion mass spectrometry (TOF-SIMS) provides unique capabilities for elemental and molecular compositional analysis of a wide variety of surfaces. This relatively new technique is finding increasing applications in analyses concerned with determining the chemical composition of various polymer surfaces, identifying the composition of organic and inorganic residues on surfaces and the localization of molecular or structurally significant secondary ions signals from biological tissues. TOF-SIMS analyses are typically performed under low primary ion dose (static SIMS) conditions and hence the secondary ions formed often contain significant structural information.This paper will present an overview of current TOF-SIMS instrumentation with particular emphasis on the stigmatic imaging ion microscope developed in the authors’ laboratory. This discussion will be followed by a presentation of several useful applications of the technique for the characterization of polymer surfaces and biological tissues specimens. Particular attention in these applications will focus on how the analytical problem impacts the performance requirements of the mass spectrometer and vice-versa.

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Optics-Free Visualization of Proteins in Single Cells by Time of Flight-Secondary Ion Mass Spectrometry Coupled with Genetically Encoded Chemical Tags

10.26434/chemrxiv.12046125 ◽

2020 ◽

Author(s):

Feifei Jia ◽

Jie Wang ◽

Yanyan Zhang ◽

Qun Luo ◽

Luyu Qi ◽

...

Keyword(s):

Mass Spectrometry ◽

Secondary Ion Mass Spectrometry ◽

Single Cells ◽

Time Of Flight ◽

E Coli ◽

Tof Sims ◽

Ion Mass Spectrometry ◽

Chemical Tags ◽

Secondary Ion

In situ visualization of proteins of interest at single cell level is attractive in cell biology, molecular biology and biomedicine, which usually involves photon, electron or X-ray based imaging methods. Herein, we report an optics-free strategy that images a specific protein in single cells by time of flight-secondary ion mass spectrometry (ToF-SIMS) following genetic incorporation of fluorine-containing unnatural amino acids as a chemical tag into the protein via genetic code expansion technique. The method was developed and validated by imaging GFP in E. coli and human HeLa cancer cells, and then utilized to visualize the distribution of chemotaxis protein CheA in E. coli cells and the interaction between high mobility group box 1 protein and cisplatin damaged DNA in HeLa cells. The present work highlights the power of ToF-SIMS imaging combined with genetically encoded chemical tags for in situ visualization of proteins of interest as well as the interactions between proteins and drugs or drug damaged DNA in single cells.

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