Analytical and Bioanalytical Chemistry

, Volume 409, Issue 14, pp 3551–3560 | Cite as

Bioanalysis of farnesyl pyrophosphate in human plasma by high-performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry and hybrid quadrupole Orbitrap high-resolution mass spectrometry

  • Hiroshi Sugimoto
  • Mie Iguchi
  • Fumihiro Jinno
Research Paper


The isoprenoids farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP) are pivotal intermediates for cholesterol homeostasis and cell signaling in the mevalonate pathway. We developed a sensitive and selective high-performance liquid chromatography tandem triple quadrupole mass spectrometry (LC-QQQ-MS) method for FPP in human plasma without the need for a derivatization process. We optimized the sample preparation procedure to extract FPP and 13C5-FPP (as internal standard) from sample fluids using methanol. Phosphate-buffered saline was used as the surrogate matrix for the preparation of calibration curves and quality control samples. Using an XBridge C18 column (3.5 μm, 2.1 × 100-mm ID) with gradient elution composed of 10 mmol/L ammonium carbonate/ammonium hydroxide (1000:5, v/v) and acetonitrile/ammonium hydroxide (1000:5, v/v) provided the sharp peaks of FPP and 13C5-FPP in human plasma. The calibration curve ranged from 0.2 to 20 ng/mL in human plasma with acceptable intra-day and inter-day precision and accuracy. The sensitivity of this bioanalytical method was sufficient for clinical analysis. The endogenous FPP plasma concentrations in 40 human healthy volunteers ascertained by LC-QQQ-MS and high-performance liquid chromatography tandem hybrid quadrupole Orbitrap high-resolution mass spectrometry (LC-Q-Orbi-MS) were comparable. Furthermore, the endogenous GGPP in human plasma was selectively detected for the first time by LC-Q-Orbi-MS. In conclusion, a sensitive bioanalytical method for FPP in human plasma by means of LC-QQQ-MS and LC-Q-Orbi-MS was developed in this study. Taking into account the versatility of LC-Q-Orbi-MS, the simultaneous detection of FPP and GGPP may be feasible in clinical practice.


Farnesyl pyrophosphate Geranylgeranyl pyrophosphate Surrogate matrix LC-QQQ-MS LC-Q-Orbi-MS 



We are thankful to Mr. Koichi Iida at Drug Metabolism and Pharmacokinetics Research Laboratories, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, for fruitful discussion about the application of high-resolution mass spectrometry for the quantification of mevalonate-derived isoprenoid intermediates. We appreciate Mr. Shio Watanabe at the Application Group, LC-MS Chromatography & MS Department, Thermo Fisher Scientific, Japan, for his excellent analytical support with LC-Q-Orbi-MS.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  1. 1.Drug Metabolism and Pharmacokinetics Research Laboratories, Pharmaceutical Research DivisionTakeda Pharmaceutical Company LimitedFujisawaJapan

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