Quantitative study of nine main components in Yueju Tablets by HPLC coupled with wavelength switching and gradient elution method / 中草药

ABSTRACT

Objective: To establish HPLC coupled with wavelength switching and gradient elution method (HPLC-DVD) for simultaneous determination of nine main components (α-cyperone, genipin-1-β-D-gentiobioside, geniposide, crocin I, senkyunolide H, senkyunolide I, senkyunolide A, ligustilide, and atractylodin) in Yueju Tablets. Methods: The chromatographic separation was achieved on Zorbax Eclipse Plus C18 (250 mm × 4.6 mm, 5 μm) column with methanol-acetonitrile (2∶1) (A)-0.2% glacial acetic acid solution (B) as mobile phases for gradient elution, at the flow rate of 0.9 mL/min; The detection wavelength was set at 240 nm for α-cyperone, genipin-1-β-D-gentiobioside and geniposide, 440 nm for crocin I, 280 nm for senkyunolide H, senkyunolide I, senkyunolide A and ligustilide, and 340 nm for atractylodin. The volume of sample injection was 10 μL. Results: The nine active components were well separated and showed good linearity, such as α-cyperone 2.58—51.60 μg/mL (r = 0.999 4), genipin-1-β-D- gentiobioside 11.99—239.80 μg/mL (r = 0.999 9), geniposide 17.96—359.20 μg/mL (r = 0.999 6), crocin-I 3.98—79.60 μg/mL (r = 0.999 7), senkyunolide H 2.82—56.40 μg/mL (r = 0.999 9), senkyunolide I 2.38—47.60 μg/mL (r = 0.999 9), senkyunolide A 6.04—120.80 μg/mL(r = 0.999 5), ligustilide 7.98—159.60 μg/mL (r = 0.999 3), and atractylodin 6.51—130.20 μg/mL (r = 0.999 2). The precision was good, and RSD was not more than 1.22%. The repeatability was good, and RSD was not more than 1.75%. The stability was good in 18 h, and RSD was not more than 1.37%. The average recoveries and corresponding RSD values were 97.64% (0.98%), 99.09% (1.46%), 100.11% (1.03%), 97.87% (0.80%), 98.59% (1.19%), 96.89% (1.34%), 99.38% (0.58%), 98.50% (1.22%), and 99.71% (0.85%), respectively. The contents of 10 batches of α-cyperone, genipin-1-β-D-gentiobioside, geniposide, crocin I, senkyunolide, H, senkyunolide I, senkyunolide A, ligustilide and atractylodin were 0.282—0.344, 2.099—2.445, 3.628—4.225, 0.758—0.913, 0.241—0.286, 0.217—0.266, 1.077—1.291, 1.386—1.623, and 1.137—1.434 mg/tablet. Conclusion: HPLC coupled with wavelength switching and gradient elution method has been established for simultaneous determination of nine components in Yueju Tablets. The method is simple, quick, accurate, and it can be used for content determination and quality control of Yueju Tablets.