Simultaneous Determination of Reserpine, Rescinnamine, and Yohimbine in Human Plasma by Ultraperformance Liquid Chromatography Tandem Mass Spectrometry

Abstract EN

A sensitive and selective UPLC-MS/MS method was developed and validated for the determination of three indolic alkaloids (reserpine, rescinnamine, and yohimbine) in human plasma using papaverine as internal standard (IS).

After a one step protein precipitation with acetonitrile, separation was carried out using C18 column (50 × 2.1 mm, i.d.

1.7 μm) and mobile phase consisting of acetonitrile : water : formic acid (60 : 40 : 0.1%, v/v/v) pumped at a flow rate of 0.2 mL/min.

The mass spectrometric determination was carried out using an electrospray interface operated in the positive mode with multiple reaction monitoring (MRM) mode.

The precursor to product ion transitions of m/z 609.32 > 195.01, m/z 635.34 > 221.03, m/z 355.19 > 144, and m/z 340.15 > 202.02 were selected for the quantification of reserpine, rescinnamine, yohimbine, and IS, respectively.

The analytical response was found to be linear in the range of 0.36–400, 0.27–300, and 0.23–250 ng/mL with lower limit of quantification of 0.36, 0.27, and 0.23 ng/mL for reserpine, rescinnamine, and yohimbine, respectively.

Validation was made following official guidelines.

The proposed method enabled reproducible results and hence could be reliable for pharmacokinetic and toxicological analysis.