Objective: To establish a fingerprint and a method for the determination of caffeic acid, rosmarinic acid, luteolin, apigenin, negletein, carvacrol and thymol in Moslae Herba using high performance liquid chromatography (HPLC). Methods: HPLC analysis was performed on an Eclipse XDB-C18 column (250 mm×4.6 mm, 5 μm) with the gradient elution of 0.1% formic acid solution (A) and acetonitrile (B) (0-17 min, 12%B→20%B; 17-30 min, 20%B→30%B; 30-40 min, 30%B→45%B; 40-55 min, 45%B→54%B). The flow rate was 1.0 mL·min-1, and the detection wavelengths were 280 nm and 330 nm. The column temperature was 30 ℃ and the injection volume was 5 μL. Results: HPLC fingerprint spectrum of Moslae Herba was established. A total of 14 common peaks in the fingerprint of Moslae Herba were assigned, and 5 components were identified as caffeic acid (peak No.5), rosmarinic acid (peak No.10), negletein (peak No.12), carvacrol (peak No.13) and thymol (peak No.14). The similarity of 17 batches of Moslae Herba samples were all over 0.910. The contents of the above seven components identified in 17 batches were determined, and their concentrations were in the linear ranges of 1.04-52.0 μg·mL-1 (r=0.999 3,n=), 6.0-300.0 μg·mL-1 (r=0.999 5, n=5), 0.12-5.88 μg·mL-1 (r=0.999 4, n=5), 0.12-5.84 μg·mL-1 (r=0.999 8, n=5), 2.42-120.8 μg·mL-1 (r=0.999 8, n=5), 2.24-112.0 μg·mL-1 (r=0.999 5, n=5), 10.2-510.0 μg·mL-1 (r=0.999 5, n=5), respectively. The average recoveries were 104.3%-97.2% with RSDs of 0.18%-2.2.%. The contents of caffeic acid, rosmarinic acid, luteolin, apigenin, 5,6-dihydroxy-7-methory-flavone, carvacrol, and thymol in 17 batches of samples were 0.442-1.028, 1.222-9.727, 0.017-0.093, 0.016-0.070, 0.279-1.013, 0.0659-1.673, 5.007-9.293 mg·g-1, respectively. Conclusion: The proposed specific HPLC fingerprint spectrum and quantitation method of seven components of Moslae Herba is simple, stable and repeatable, which can provide a reference for quality evaluation of Moslae Herba.
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