Stand out from matrix: Ultra-sensitive LC-MS/MS method for determination of histamine in complex biological samples...

Area of Science:

• Biochemistry
• Analytical Chemistry
• Clinical Diagnostics

Background:

• Histamine is a key endogenous component involved in various physiological processes.
• Accurate histamine quantification is vital for diagnosing conditions such as allergies, asthma, and anaphylactic shock.
• Detecting low histamine concentrations in complex biological samples presents significant analytical challenges due to stability and matrix interference.

Purpose of the Study:

• To develop an ultra-sensitive and accurate liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for histamine quantification.
• To overcome the challenges associated with detecting low abundant histamine in clinical samples.
• To establish a reliable method for histamine determination in complex biological matrices.

Main Methods:

• Development of an LC-MS/MS method incorporating derivatization with diisopropyl phosphite (DIPP).
• Application of isotope dilution and solid-phase extraction (SPE) for enhanced sensitivity and interference removal.
• Optimization of derivatization to improve LC retention and ionization efficiency.

Main Results:

• Achieved an ultra-sensitive quantification method with a lowest limit of quantification (LLOQ) of 0.1 pg/mL.
• Demonstrated successful application in determining histamine in low nanoliter serum volumes and single cells.
• Validated the method by quantifying histamine in clinical serum samples from asthma patients and treated mast cells.

Conclusions:

• The developed LC-MS/MS method provides unprecedented sensitivity and accuracy for histamine determination.
• This approach enables the precise measurement of histamine in challenging clinical samples, aiding in disease diagnosis.
• The method is suitable for analyzing trace levels of histamine in various biological matrices, including serum and cellular samples.