Summary
Isolation and Procedure for Simultaneous Quantitative Determination of Imperatorin and Isoimperatorin in Angelicae dahuricae Radix by HPLC
In order to provide a method for quality control of Radix Angelicae dahuricae and preparations containing this Radix, this paper presents the isolation of the major coumarins from A.dahurica root and using them in method development for simultaneous quantification of these compounds in raw material and preparations by means of HPLC-PDA.
From roots of Angelica dahurica, two coumarins, i.e., imperatorin (345 mg) and isoimperatorin (962 mg) were isolated. Their structure were confirmed by MS, NMR data compared with those reported in literatures. Their purity was estimated to be 100% by means of HPLC-PDA.
The standard solution of imperatorin and isoimperatorin showed the linearity in the range of 0.05 - 50 µg/ml (R2 @ 1.0000) and 0,1 - 50 µg/ml (R2 = 0.9999), respectively. The repeatability of imperatorin and isoimperatorin: 0.086% (RSD 0.002) and 0.031% (RSD 0.0004%). The average of recoveries were 96.77%, RSD 0,21 for imperatorin and 95.28%, RSD 1,32 for isoimperatorin. LODs and LOQs were 0.1 and 0.3 µg/ml for imperatorin, 0.25 and 0.8 µg/ml for isoimperatorin. The repeatability of sample preparations were with RSD 0.58 and RSD 0.08, respectively. Application of the method to analysis of 6 A. dahurica root samples (M1 - M6) and 4 preparations (S1 - S4) containing A. dahurica distributed on the HCMC market showed that: M1, M2, S4 samples were conformed to the proposed standard for imperatorin and isoimperatorin; other M samples and S1, S3 meet quantitative requirements of the standard. The preparation S2 did not correspond.
The procedures for isolation and quantification of imperatorin and isoimperatorin were established.
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