Simultaneous Determination of Aesculin, Aesculetin, Fraxetin, Fraxin and Polydatin in Beagle Dog Plasma by UPLC-ESI-MS/MS and Its Application in a Pharmacokinetic Study after Oral Administration Extracts of Ledum palustre L.

Abstract

A rapid, simple and sensitive ultra-performance liquid chromatography-electrospray-ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) method was developed and validated for the simultaneous determination of aesculin, aesculetin, fraxetin, fraxin and polydatin in beagle dog plasma for the first time. Plasma samples were pretreated by protein precipitation with methanol. Chromatographic separation was performed on an Acquity UPLC HSS T3 C18 column (2.1 mm × 100 mm, 1.8 μm) with gradient elution at a flow rate of 0.4 mL/min, using a mobile phase consisting of 0.1% formic acid (A) and acetonitrile (B). The analytes and IS were detected by multiple reaction monitoring (MRM) via negative ion mode with ion transitions of m/z 339.1–m/z 176.8 for aesculin, m/z 176.8–m/z 88.9 for aesculetin, m/z 206.8–m/z 192.1 for fraxetin, m/z 369.1–m/z 206.9 for fraxin, m/z 389.1–m/z 227.0 for polydatin and m/z 415.2–m/z 295.1 for puerarin. This method was validated according to the FDA guidelines and the results met the requirements of analysis. The calibration curves of analytes were linear with correlation coefficients more than 0.9980. The intra- and inter-day precisions were less than 15% and the accuracy was within ±15%. The maximum plasma concentration (Cmax) of aesculin, aesculetin, fraxetin, fraxin and polydatin was 46.75 ± 7.46, 209.9 ± 57.65, 369.7 ± 48.87, 67.04 ± 12.09 and 47.14 ± 12.04 ng/mL, respectively. The time to reach the maximum plasma concentration (Tmax) was 1.32 ± 0.38 h for aesculin, 1.03 ± 0.27 h for aesculetin, 0.94 ± 0.23 h for fraxetin, 0.83 ± 0.18 h for fraxin and 1.15 ± 0.15 h for polydatin. The results indicated that the absorption of aesculin might be slow in beagle dog plasma. This method was successfully applied for pharmacokinetics in beagle dog plasma after oral administration of the extracts of Ledum palustre L. at a dosage of 0.27 g/kg.

Simultaneous Determination of 78 Compounds of Rhodiola rosea Extract by Supercritical CO2-Extraction and HPLC-ESI-MS/MS Spectrometry

“…A total of 300 peaks were detected in the chromatogram. By comparing the m/z values, the RTand the fragmentation patterns with the MS 2 spectral data taken from the literature [2,17,[35][36][37][38][39][40][41][42][43][44][45][46][47][48][49][50] or to search the data bases (MS2T, MassBank, HMDB). 78 metabolites were putatively identified as phenols, aromatic compounds, phenyl alkanoids, flavonoids, monoterpenoids, acyclic alcohol glycosides, anthocyanins etc.…”

In Vivo Pharmacokinetic Analysis Utilizing Non-Targeted and Targeted Mass Spectrometry and In Vitro Assay against Transient Receptor Potential Channels of Maobushisaishinto and Its Constituent Asiasari Radix

“…Analytical methods (e.g., LC mobile phase, column, and mass parameters) in association with the non-targeted analysis were selected based on our experiences as well as previous report [ 42 ]. To enable the non-targeted analysis to detect more extensive compounds in the plasma, the pretreatment method was selected to be the deproteination method.…”

Simultaneous Quantification of Five Sesquiterpene Components after Ultrasound Extraction in Artemisia annua L. by an Accurate and Rapid UPLC–PDA Assay

“…The precision of the UPLC method was evaluated by using a mixed standard solution of marker compounds at low, medium and high levels to measure intra-day and inter-day variations [27]. The mixed standard solution was analyzed five times in a single day to determine intra-day precision, and the inter-day precision of the method was evaluated by repeating the injection of the standard solution for three consecutive days.…”