A facile and sensitive HPLC-MS/MS method for quantification of 3,3'-diindolylmethane in plasma samples

Area of Science:

• Biomedical research
• Analytical chemistry
• Pharmacokinetics

Background:

• Indole-3-carbinol and its metabolite 3,3'-diindolylmethane (DIM) show potential antiatherogenic, anticarcinogenic, and antioxidant effects.
• Biological activity of DIM is observed at ng/ml concentrations, necessitating sensitive analytical methods.
• Quantifying DIM in biological samples is crucial for understanding its therapeutic potential.

Purpose of the Study:

• To develop and validate a highly sensitive HPLC-MS/MS method for quantifying DIM in human plasma.
• To establish a reliable method for bioanalytical studies involving DIM.
• To support pharmacokinetic studies of DIM and related compounds.

Main Methods:

• Developed an HPLC-MS/MS method for DIM quantification in human plasma.
• Validated the method according to European Medicines Agency guidelines.
• Utilized acetonitrile-induced protein precipitation with ammonium sulfate for sample preparation.
• Employed a Synergi Fusion-RP C18 column with a mobile phase of acetonitrile and water (0.1% formic acid; 85:15, v/v).
• Used atmospheric pressure chemical ionization with multiple reaction monitoring (MRM) in positive polarity mode.

Main Results:

• The method achieved good sensitivity, selectivity, accuracy, and precision.
• Quantification range was 5-500 ng/ml, with a limit of detection of 1 ng/ml and a lower limit of quantification of 5 ng/ml.
• The total analysis time was 3.20 minutes.
• Successfully applied the method to analyze plasma samples from human volunteers and rabbits after oral administration of DIM-containing drugs or DIM itself.

Conclusions:

• The developed HPLC-MS/MS method is sensitive, selective, and efficient for quantifying DIM in human plasma.
• This validated method is suitable for high-throughput bioanalysis and pharmacokinetic studies.
• The method facilitates further research into the biomedical applications of DIM.