Indole-3-carbinol (I3C) is a dietary constituent derived from glucobrassicin, found in cruciferous vegetables. This compound is the subject of on-going biomedical research due to its possible antiatherogenic, anticarcinogenic, and antioxidant effects. The antitumor properties are mainly associated with its main metabolite — 3.3'-diindolylmethane (DIM). It is known, that the biological activity of the chemical compound is manifested in the concentration range of ng/mL. Currently, there are no validated analytical methods that allowed the determination of I3C and DIM in various human biological samples. Therefore, the aim of the present study was to develop and validate a simple and highly sensitive HPLC—MS/MS method for simultaneous quantification of I3C and DIM in human plasma. The acetonitrile-induced protein precipitation method was adapted for extracting the analytes from human plasma. The separation was conducted using Macherey—Nagel Nucleodur C8 ec column at 40°C under gradient elution with 5 mM ammonium formate in water (A) and acetonitrile (B) mobile phases. The flow rate was 0.50 mL/min with a total analysis run time of 10.5 min. I3C and DIM were eluted at 4.81 ± 0.03 and 8.71 ± 0.03 min, respectively. 4-Methoxy-1-methylindole was used as an internal standard. The analytes were monitored using multiple reaction monitoring scans in positive polarity mode. The ion transitions were at m/z 130.2 ^ 77.2 (I3C), 247.2 ^ 130.2, and 130.2 ^ 77.2 (DIM). The method showed good linearities for I3C and DIM within the range of 20-500 ng/mL and 5—500 ng/mL, respectively. Conclusively, the method has been successfully applied to the analysis of volunteers ’ plasma samples after a single and multiple oral administration of a drug containing I3C as the active substance.
Перспективы развития фундаментальных наук. Т. 2 : сборник научных трудов XVIII Международной конференции студентов, аспирантов и молодых ученых, 27–30 апреля 2021 г.. Томск, 2021. Т. 2 : Химия. С. 140-142
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Химический факультет
Совместное определение индол-3-карбинола и 3.3'-дииндолилметана в плазме крови человека методом ВЭЖХ-МС/МС