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Simultaneous Quantitation of Isoprenoid Pyrophosphates in Plasma and Cancer Cells Using LC-MS/MS.
Chhonker, Yashpal S; Haney, Staci L; Bala, Veenu; Holstein, Sarah A; Murry, Daryl J.
Afiliación
  • Chhonker YS; Department of Pharmacy Practice, University of Nebraska Medical Center, Omaha, NE 68198, USA. y.chhonker@unmc.edu.
  • Haney SL; Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA. staci.haney@unmc.edu.
  • Bala V; Department of Pharmaceutical Sciences, Mohanlal Sukhadia University, Udaipur, Rajasthan 313001, India. veenu2bala@gmail.com.
  • Holstein SA; Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA. sarah.holstein@unmc.edu.
  • Murry DJ; Fred and Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, NE 68198, USA. sarah.holstein@unmc.edu.
Molecules ; 23(12)2018 Dec 11.
Article en En | MEDLINE | ID: mdl-30544938
Isoprenoids (IsoP) are an important class of molecules involved in many different cellular processes including cholesterol synthesis. We have developed a sensitive and specific LC-MS/MS method for the quantitation of three key IsoPs in bio-matrices, geranyl pyrophosphate (GPP), farnesyl pyrophosphate (FPP), and geranylgeranyl pyrophosphate (GGPP). LC-MS/MS analysis was performed using a Nexera UPLC System connected to a LCMS-8060 (Shimadzu Scientific Instruments, Columbia, MD) with a dual ion source. The electrospray ionization source was operated in the negative MRM mode. The chromatographic separation and detection of analytes was achieved on a reversed phase ACCQ-TAG Ultra C18 (1.7 µm, 100 mm × 2.1 mm I.D.) column. The mobile phase consisted of (1) a 10 mM ammonium carbonate with 0.1% ammonium hydroxide in water, and (2) a 0.1% ammonium hydroxide in acetonitrile/methanol (75/25). The flow rate was set to 0.25 mL/min in a gradient condition. The limit of quantification was 0.04 ng/mL for all analytes with a correlation coefficient (r2) of 0.998 or better and a total run time of 12 min. The inter- and intra-day accuracy (85⁻115%) precision (<15%), and recovery (40⁻90%) values met the acceptance criteria. The validated method was successfully applied to quantitate basal concentrations of GPP, FPP and GGPP in human plasma and in cultured cancer cell lines. Our LC-MS/MS method may be used for IsoP quantification in different bio-fluids and to further investigate the role of these compounds in various physiological processes.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Fosfatos de Poliisoprenilo / Sesquiterpenos / Cromatografía Liquida / Espectrometría de Masas en Tándem Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Molecules Asunto de la revista: BIOLOGIA Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Suiza
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Fosfatos de Poliisoprenilo / Sesquiterpenos / Cromatografía Liquida / Espectrometría de Masas en Tándem Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Molecules Asunto de la revista: BIOLOGIA Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Suiza