A rapid and sensitive LC‐MS/MS method for the determination of linarin in small‐volume rat plasma and tissue samples and its application to pharmacokinetic and tissue distribution study

Abstract

A rapid and sensitive LC-MS/MS method was developed for the determination of linarin in small-volume rat plasma and tissue sample. Sample preparation was employed by the combination of protein precipitation (PPT) and liquid-liquid extraction (LLE) to allow measurement over a 5-order-of-magnitude concentration range. Fast chromatographic separation was achieved on a Hypersil Gold column (100 × 2.1 mm i.d., 5 µm). Mass spectrometric detection was achieved using a triple-quadrupole mass spectrometer equipped with an electrospray ionization interface operating in positive ionization mode. Quantification was performed using selected reaction monitoring of precursor-product ion transitions at m/z 593 → 285 for linarin and m/z 447 → 271 for baicalin (internal standard). The total run time was only 2.8 min per sample. The calibration curves were linear over the concentration range of 0.4-200 µg/mL for PPT and 0.001-1.0 µg/mL for LLE. A lower limit of quantification of 1.0 ng/mL was achieved using only 20 μL of plasma or tissue homogenate. The intra- and inter-day precisions in all samples were ≤14.7%, while the accuracy was within ±5.2% of nominal values. The validated method has been successfully applied to pharmacokinetic and tissue distribution study of linarin.

The Phenolic Fraction of Mentha haplocalyx and Its Constituent Linarin Ameliorate Inflammatory Response through Inactivation of NF-κB and MAPKs in Lipopolysaccharide-Induced RAW264.7 Cells

“…It has also been reported that linarin possesses analgesic, antipyretic [ 46 ], and neuroprotective activities [ 47 ], and showed a protective effect against d -glactosamine/LPS-induced fulminant hepatic failure [ 48 ]. Furthermore, a tissue distribution study of linarin has revealed that it was rapidly and widely distributed in the heart, liver, spleen, lung, kidney, brain, muscle, stomach, small intestine, epididymis, pancreas, and bladder of rats after intragastric administration [ 49 ]. Since there was little data on the anti-inflammatory mechanism of linarin [ 50 ], the present study explored its involvement in the anti-inflammatory action of the MHP extract.…”

Technologies to improve the sensitivity of existing chromatographic methods used for bioanalytical studies

“…Protein precipitation (PPT) is widely used in biological analysis, especially for high-throughput screening, due to its fast, simple, and cost-efficient characteristics (Feng, Liu, Wang, & Di, 2016;Zhao et al, 2019). Moreover, PPT often provides higher recovery of analytes when compared with solid-phase extraction (SPE) and liquid-liquid extraction (LLE) after optimizing the most appropriate precipitation reagent (Rathod et al, 2017;Wang et al, 2018).…”

LC–MS/MS Method for Simultaneous Determination of Linarin and Its Metabolites in Rat Plasma and Liver Tissue Samples: Application to Pharmacokinetic and Liver Tissue Distribution Study After Oral Administration of Linarin

“…However, one of our previous studies revealed that linarin shows low bioavailability (0.47%) after oral administration, which suggests that linarin could be extensively metabolized after oral administration, and the metabolites of linarin could still show various pharmacological activities [11]. Acacetin, apigenin, and p -hydroxy benzaldehyde (Figure 1) have been identified as the metabolites of linarin [12].…”

“…We found that these three metabolites showed a protective effect against severe d -GalN-induced hepatic failure in rats (data not shown). Up to now, the pharmacokinetic study of linarin had been extensively reported [11,13,14,15,16]; however, no pharmacokinetic study of its metabolites is available. Thus, further studies to elucidate the pharmacokinetics and liver tissue distribution of linarin and its active metabolites are still needed and will be helpful for a better understanding of the pharmacological effects of linarin.…”