Abstract
Guang Dilong is a Traditional Chinese Medicine prepared from the dried body of Pheretima aspergillum (E. Perrier), a species of earthworm. However, preparations of Guang Dilong may be adulterated by other species and a method of quality control is needed. A method was developed to analyze and authenticate commercial Guang Dilong, utilizing ultra-high performance liquid chromatography (UHPLC) coupled with diode array detection (DAD). Equipment included an Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm). The mobile phase consisted of acetonitrile and 0.01% formic acid, pumped at 0.3 mL/min. Wavelength detection was at 260 nm. Twenty-two batches of confirmed P. aspergillum samples (reference) from different sources and 20 batches of adulterated samples were analyzed to establish a reference fingerprint for commercial Guang Dilong. Five peaks in the fingerprints of the reference batches were identified as characteristic; six characteristic peaks in the fingerprints of the adulterants were identified by comparing their retention time with those of the references. The total 42 batches of samples were compared with the reference fingerprint, and the fingerprints of the P. aspergillum samples were similar. The UHPLC-DAD method can simultaneously determine the contents of six compounds (hypoxanthine, xanthine, uridine, inosine, guanosine, and adenosine) in the reference and adulterated batches. The six compounds showed good regression (r > 0.9999) within test ranges. The recovery (accuracy) was 98.25 to 101.68%, with relative standard deviation <2.67%. In summary, this UHPLC-DAD method combines chromatographic fingerprint with quantification analysis and could be readily used as an efficient quality control method for Guang Dilong.
Highlights
Guang Dilong is a Traditional Chinese Medicine (TCM) consisting of the dried body of Pheretima aspergillum
After comparison with the GenBank database, the homology of the gene sequences for sample batches No 1-No 22 was above 98%, confirming that the to-be-detected tissues were P. aspergillum (Perrier, 1872) [26] (Table 1, Figure 2)
The results indicated that this HPLC-diode array detection (DAD) method accurately determined six chemical markers in the samples of commercial Guang Dilong
Summary
Guang Dilong is a Traditional Chinese Medicine (TCM) consisting of the dried body of Pheretima aspergillum The animal is mainly distributed in Guangdong, Guangxi, and Hainan provinces. Modern research has indicated that Guang Dilong exerts many pharmacological effects, such as decreasing blood pressure, preventing thrombus formation, and preventing cancer [2]. Certain preparations of Guang Dilong have been approved for clinical use in China [3]
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