To establish an HPLC characteristic chromatogram for Hyssopus cuspidatu Boriss. and control quality and identity authentication for the herb.

The HPLC method was used, and the mobile phases were acetonitrile (A) and 0.1% aqueous phosphoric acid (B) in gradient elution(0-15 min, 10% A; 15-30 min, 10% A→15% A; 30-40 min, 15% A→18% A; 40-60 min, 18% A→23% A; 60-70 min, 23% A→55% A; 70-85 min, 55% A→60% A; 85-85.1 min, 60% A→10% A; 85.1-90 min, 10% A). The column temperature was 30 ℃, the flow rate was 1.0 mL·min^-1, the detection wavelength was 330 nm and the injection volume was 20 μL. Fifteen batches of Hyssopus cuspidatu Boriss. and its adulterants, notably Cynanchum officinale, were analyzed to construct characteristic chromatogram. The authenticity and quality of Hyssopus cuspidatu Boriss. were assessed through principal component analysis(PCA), cluster analysis, and similarity analysis.

Analysis of the characteristic chromatogram of ten Hyssopus cuspidatu Boriss. batches revealed fifteen distinct peaks, with six compounds identified as isodendrobioside, molluscoside, rosmarinic acid, salvianolic acid, geranylgeranyl, and trichothecenes. Cluster analysis segregated the Hyssopus cuspidatu Boriss. samples into three distinct groups, primarily influenced by geographical (latitude and longitude) and environmental (climate) factors. PCA results indicated that four principal components accounted for 92.914% of the variance. The similarities for the ten batches of Hyssopus cuspidatu Boriss. ranged from 0.896 to 0.997, with no significant difference observed between the wild and cultivated varieties in terms of overall HPLC profile and relative peak content. The overall profiles of the HPLC characteristic chromatogram and the contents of the main components of the Hyssopus cuspidatu Boriss. and the Nepeta bracteata Benth. were significantly different.

The HPLC characteristic chromatogram method developed in this study is reproducible, stable, and practical for the quality control and authentication of Hyssopus cuspidatu Boriss. Moreover, it offers a reliable analytical technique for the quality evaluation and authentication of this herb, providing substantial data support.