The HPLC Fingerprint and Isovanillin Content of Benincasa hispida Seeds

The Benincasa hispida seed is used as traditional Chinese medicine (TCM). However, there is still a lack of medicinal quality control of B. hispida seeds. The seeds may contain isovanillin, but this finding remains to be confirmed and quantified. The current study aimed to confirm the existence of isovanillin and then preliminarily establish medicinal quality standards for B. hispida seeds. Fourteen batches of unilateral and bilateral B. hispida seeds were purchased from 7 different producers in China. Semipreparative high-performance liquid chromatography (HPLC) was used to isolate and purify the isovanillin from B. hispida seeds. Its chemical structure was elucidated by UV, 1H-NMR, IR spectroscopy, and GC-MS. The B. hispida seed fingerprint and isovanillin determination were performed on an HPLC instrument. Data obtained from the unilateral and bilateral specimens were analyzed with a similarity evaluation system. The HPLC fingerprint showed 19 characteristic peaks with high similarity between the unilateral and bilateral B. hispida seeds. The isovanillin content among the fourteen batches ranged from 13.46 to 46.80 μg/g. The results of this study may provide a preliminary reference for the quality control of B. hispida seeds.