Quantitative and qualitative analysis of common peaks in chemical fingerprint of Yuanhu Zhitong tablet by HPLC-DAD-MS/MS

Abstract

A quality control (QC) strategy for quantitative and qualitative analysis of "common peaks" in chemical fingerprint was proposed to analyze Yuanhu Zhitong tablet (YZT), using high performance liquid chromatography with diode array detector and tandem mass spectrometry (HPLC-DAD-MS/MS). The chromatographic separation was achieved on an Agilent Eclipse plus C₁₈ column with a gradient elution using a mixture of 0.4‰ ammonium acetate aqueous (pH 6.0 adjusted with glacial acetic acid) and acetonitrile. In chemical fingerprint, 40 peaks were assigned as the "common peaks". For quantification of "common peaks", the detection wavelength was set at 254 nm, 270 nm, 280 nm and 345 nm, respectively. The method was validated and good results were obtained to simultaneously determine 10 analytes (protopine, jatrorrhizine, coptisine, palmatine, berberine, xanthotoxin, bergapten, tetrahydropalmatine, imperatorin and isoimperatorin). For qualification of "common peaks", 33 compounds including 10 quantitative analytes were identified or tentatively characterized using LC-MS/MS. These results demonstrated that the present approach may be a powerful and useful tool to tackle the complex quality issue of YZT.

Keywords: Alkaloids; Coumarins; HPLC-DAD–MS/MS; Quality control; Yuanhu Zhitong tablet.

Fig. 1

Chemical structures of 33 identified compounds in Yuanhu Zhitong tablet (YZT).

Fig. 2

Representative HPLC-DAD chromatograms of mixed standard solutions (A) at 254 nm, 270 nm, 280 nm and 345 nm; YZT (B) at 254 nm, 270 nm, 280 nm and 345 nm; the negative sample without Radix Corydalis (C) at 280 nm; and the negative sample without Rhizoma Angelicae dahuricae (D) at 280 nm. (3) protopine; (7) jatrorrhizine; (8) coptisine; (14) palmatine; (15) berberine; (20) xanthotoxin; (23) bergapten; (28) tetrahydropalmatine; (37) imperatorin; (40) isoimperatorin.

Fig. 3

HPLC–ESI-MS total ion chromatogram (TIC) in positive ion mode of (A) the mixed standard and (B) YZT.

Fig. 4

The chromatogram of the investigated 12 samples of YZT.