Simultaneous determination of 19 constituents in the Jian‐Nao‐Ning mixture using high‐performance liquid chromatography‐tandem mass spectrometry

Jin‐Nao‐Ning mixture (JNNm) has been clinically used to treat insomnia in China. However, the major constituents of JNNm were not revealed. In this study, an ultra‐high‐performance liquid chromatography‐tandem mass spectrometry was established and validated for simultaneous quantitation of 19 representative constituents of JNNm. The method had good specificity and adequate sensitivity. The calibration curves of the constituents displayed good linearity (r > 0.990) in appropriate ranges. The intra‐ and inter‐day precision at three quality control (QC) levels ranged from 0.68% to 5.17% and 0.42% to 5.37%, respectively. The recovery ranged from 91.76% to 110.06%, with a precision of less than 5.07%. In addition, the method had good repeatability (relative standard deviation [RSD] < 4.93%). The analytes were all stable for 24 h (RSD < 7.98%) at room temperature. The results of the quantitative analysis showed that salvianolic acid B (495.7–606.3 μg/mL) is the most abundant constituent, followed by betaine (411.3–468.3 μg/mL), salvianic acid A (10.7–34.9 μg/mL), lobetyolin (11.6–14.6 μg/mL), spinosin (4.0–7.6 μg/mL), schisandrol A (1.5–4.1 μg/mL), and schisandrol B (1.2–1.5 μg/mL). Concentrations of other constituents were below 1 μg/mL. This study would be helpful for the QC of JNNm and the revelation of its effective constituents.