Application of a high-resolution benchtop quadrupole-Orbitrap mass spectrometry for the pharmacokinetics and tissue distribution of Danhong injection in guinea pig

• Simultaneous quantitation of four analytes of Danhong injection in guinea pig using Q-Orbitrap HRMS.
• Q-Orbitrap HRMS in full MS/dd-MS2 mode was used for screening of salvianic acid A sodium, protocatechualdehyde, salvianolic acid B and rutin.
• Pharmacokinetics and tissue distribution of four analytes of Danhong injection were detected in guinea pig.

Danhong injection (DHI) is a compound preparation of traditional Chinese medicine Salviae miltiorrhiza (a.k.a. Danshen) and Flos Carthami tinctorii. Phenolic acids, such as salvianic acid A sodium (SAS), protocatechualdehyde (PAL), salvianolic acid B (SAB) and flavonoids, such as rutin, are the main bioactive ingredients of DHI. This paper presents an application of ultra high-performance liquid chromatography and quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HR MS) to quantify salvianic acid A sodium, protocatechualdehyde, salvianolic acid B, and rutin in guinea pig plasma and tissues for pharmacokinetic and tissue distribution studies. Biological samples were processed with methanol extraction, and calibration curves were determined with indometacin as internal standard (I.S.). The analytes were separated by a Hypersil GOLD aQ C18 column and detected using negative ion full MS/dd-MS2 (data-dependent MS2) mode. The full MS scan acquired data for identification and quantification, and dd-MS2 scan obtained product ion spectra for confirmation. The response showed good linear relationship over the concentration range between 10.22 and 5110 ng/mL for salvianic acid A sodium, 10.06–5030 ng/mL for salvianolic acid B, 11.12–5560 ng/mL for protocatechualdehyde, and 10.58–5290 ng/mL for rutin, all the coefficients of correlation (r2) >0.9990. The lower limit of quantification (LLOQ) was 10 ng/mL for the analytes. Overall, the novel UHPLC-Q-Orbitrap has demonstrated great performance for rapidity, accuracy, high sensitivity, and high selectivity. It was successfully applied to the pharmacokinetic and tissue distribution studies of Danhong injection. These acquired data would provide helpful information for the clinical applications, mechanism studies, and technology promotion of DHI in the future.

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