Development and Validation of a HPLC-UV Method with Pre-column Derivatization for Determination of Cinnabar in Jufang Zhibao Pills

Xin Zhang, Jia Yao, Ying Ren, Yuan Li, Zhimin Xie


In this work, a reliable and accurate high-performance liquid chromatography method with
pre-column derivatization was established and validated for determination of cinnabar in
Jufang Zhibao pills. Scanning electron microscope (SEM) image was used to identify the
types of cinnabar crude drug in Jufang Zhibao pills. The chromatography separation was
performed on a Welch XB-C18 column (250 mm × 4.6 mm, 5 μm). The mobile phase consists
of water spiked with 0.022 mmol/L sodium diethyldithiocarbamate (A, pH adjusted to 8–9 by
ammonia water) and methanol (B, 80:20, v/v) at flow rate of 1.0 ml/min with the detected
wavelength was 272 nm. The oven temperature was set at 35°C. The calibration for cinnabar
content has good linearity (R
2 =0.9999) over the range of 2.43–300 μg/ml and the average
recovery was less then 1.90%. The limits of detection and quantification were 0.1127 μg and
0.2065 μg/ml. The results indicated that the proposed method has advantages of high
accuracy, good repeatability and stability and can be successfully used for determination of
cinnabar in Jufang Zhibao pills. It provides a basis for drug manufacture quality control and
proves the feasibility of the pre-column derivatization method during the determination of
cinnabar in Jufang Zhibao pills.


Jufang Zhibao pills, Cinnabar, Prederivatization, HPLC-UV, Sodium diethyldithiocarbamate.

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