Rapid, Selective, and Sensitive Analysis of Triterpenic Acids in Hippophae rhamnoides L. Using HPLC with Pre-Column Fluorescent Derivatization and Identification with Post-Column APCI-MS

Abstract

Hippophae rhamnoides L., a fruit which is rich in natural chemical ingredients, has become an important raw material of health food, medicine, and cosmetics. In the present work, a method using 2-(5-benzoacridine)ethyl-p-toluenesulfonate as pre-column derivatization reagent with high sensitivity, selectivity, and short separation time was developed to analyze triterpenic acids (TTAs) in H. rhamnoides L. by HPLC with fluorescence detection and simultaneously confirmed by post-column atmospheric pressure chemical ionization-mass spectrometry. The method was validated by linearity, limits of detection (LODs) and lower limits of quantification (LLOQs), precision, and accuracy. Good linear correlations were observed for all TTAs with correlation coefficients greater than 0.9996. LODs and LLOQs were in the range of 1.71–2.14 and 7.45–8.23 ng mL⁻¹, respectively. The method was successfully employed to analyze TTAs in H. rhamnoides L. from various origins in the Qinghai–Tibetan plateau with trace amount of samples within 15 min. A systemic mapping of the TTAs in H. rhamnoides L. of different origins was established. The results indicated that H. rhamnoides L. is rich in TTAs, especially in oleanolic and ursolic acids, and the content of TTAs varies across origins. This work will support the research on pharmaceutical applications of H. rhamnoides L. and provide information for quality control.

Keywords:

• APCI-MS
• fluorescence detection
• Hippophae rhamnoides L.
• HPLC
• pre-column derivatization
• triterpenic acids

Notes

^a Y, peak area; X, injected amount of each TTA (pmol).

^a not detected.

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