OBJECTIVE

2

To establish a quantitative analysis of multi-components by single-marker （QAMS） method for simultaneous determination of six flavonoids and two phenolic acids in

Perilla frutescens

leaves using scutellarin and rosmarinic acid as internal reference substances， and apply this method to determine the contents of eight components in 20 batches of

P. frutescens

leaves samples from different regions.

METHODS

2

Scutellarin served as the internal reference to calculate relative correction factors （RCFs） for scutellarin-7-

O

-diglucuronide， luteolin-7-

O

-diglucuronide， apigenin-7-

O

-diglucuronide， luteolin-7-

O

-

β

-D-glucuronide and apigenin-7-

O

-glucuronide. Rosmarinic acid was employed as the internal reference to determine the RCF for caffeic acid. The contents of the above flavonoids and phenolic acids were calculated with QAMS， and compared with the results of external standard method.

RESULTS

2

The eight analytes demonstrated excellent linearity within their respective concentration ranges

（

r

≥0.999 0）. The mean recovery rates for spiked samples ranged from 95.60% to 102.15%， with relative standard deviations （RSDs） of 0.72% to 2.70% （

n

＝6）. The method exhibited good precision， repeatability， and stability （RSD＜2.50%，

n

＝6）. Variations in instruments， columns， column temperature， flow rate， and formic acid volume fraction had minimal impact on the RCFs （RSD＜3%，

n

＝3）. Comparison with the external standard method showed no significant differences in the content of each component across batches， except for caffeic acid in the ZS12 batch （absolute value of RE＜5%，

n

＝2）. The contents of six flavonoid components in

P. frutescens

leaves samples varied significantly across different geographic origins， while the content of total flavonoids showed no significant difference. In contrast， the contents of two phenolic acid components and total phenolic acid exhibited significant variation among samples from different regions.

CONCLUSIONS

2

The developed QAMS method can simultaneously determine the contents of six flavonoids and two phenolic acids in

P. frutescens

leaves. It is convenient for detection， highly accurate， and cost-effective. This method is suitable for the quality control of

P. frutescens

leaves， and the variation of flavonoid and phenolic acid content in samples from different regions provides a reference for the selection of optimal cultivation areas.