OBJECTIVE

2

To establish an ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） method for the simultaneous determination of the contents of six

Aconitum

alkaloids in Compound chondroitin sulfate tablets， and to perform quality evaluation of samples from different batches， thereby providing a reference for the quality control of this preparation.

METHODS

2

Chromatographic separation was performed on an Agilent SB-C

18

column using gradient elution with 0.2% formic acid （adjusted to pH 7.5 with ammonia） and acetonitrile as the mobile phases. The flow rate was 0.3 mL/min， the column temperature was 35 ℃， and the injection volume was 1 μL. Mass spectrometric analysis was performed using an AJS ESI

+

ion source in multiple reaction monitoring mode， with quantification by the external standard method. Method validation was performed. The established UPLC-MS/MS method was applied to determine the contents of the six

Aconitum

alkaloids in 64 batches of samples from three manufacturers， including three diester-type alkaloids （aconitine， hypaconitine， and mesaconitine） and three monoester-type alkaloids （benzoylaconine， benzoylhypaconine， and benzoylmesaconine）. Descriptive statistics， the Kruskal-Wallis

H

test， principal component analysis （PCA）， and Spearman’s rank correlation analysis were used to evaluate the content distribution of

Aconitum

alkaloids， differences among samples from different manufacturers， and correlations among the components.

RESULTS

2

Good linearity was observed for the six

Aconitum

alkaloids over their respective concentration rang

es， with correlation coefficients （all

r

＞0.99）. The relative standard deviations （RSDs） for precision， repeatability， and stability were all less than 5%. The limits of detection and quantification were no higher than 0.003 ng/mL and 0.01 ng/mL， respectively. The mean recoveries ranged from 96.43% to 102.30%， with RSDs ranging from 0.35% to 4.92%. Among the 64 batches of samples， the total content of monoester-type alkaloids （mean value of 0.034 9 mg/tablet） was markedly higher than that of diester-type alkaloids （mean value of 0.002 2 mg/tablet）. Benzoylmesaconine showed the highest content， with a mean value of 0.031 0 mg/tablet， whereas the other five components were present at relatively low levels and showed considerable inter-batch variation. No statistically significant differences were observed in the contents of the six components among samples from different manufacturers （

P

＞0.05）. PCA showed no obvious clustering trend， and significant correlations were observed among some components （

P

＜0.05）.

CONCLUSIONS

2

The established UPLC-MS/MS method enables the rapid and accurate simultaneous determination of the above-mentioned six

Aconitum

alkaloids in Compound chondroitin sulfate tablets. The quality evaluation of multiple batches of samples provides a reference for the quality control of this preparation.