Quantitative Measurement of Melittin in Asian Honeybee Venom Using a New Method Including UPLC-QqTOF-MS

Sheng Huang,Yan Wang,Chundong Liu,Zeqin Guo,Jianhua Wang

doi:10.3390/toxins12070437

Abstract

Asian honeybee venom is widely used in traditional oriental medicine. Melittin is the main component of Asian honeybee venom. In the present study, an ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QqTOF-MS) method was used for accurate qualitative and quantitative analyses of melittin in Asian honeybee venom. The results showed that the dynamic linear range of melittin was from 0.094 to 20 μg/mL, and the limit of quantification was 0.3125 μg/mL. The spiking recovery of melittin in honeybee venom ranged from 84.88% to 93.05%. Eighteen Asian honeybee venom samples in eighteen batches were collected from two different zones of China, and their melittin contents were measured. The contents of melittin in Asian honeybee venom samples was 33.9–46.23% of dry weight. This method proved a useful tool for the rapid evaluation of the authenticity and quality of Asian honeybee venom in terms of the melittin contents, and will contribute to a broader understanding of Asian honeybee venom.

Highlights

Apis cerana Fabricius is a type of Asian honeybee whose venom has been widely applied as a traditional oriental medicine to treat some diseases such as chronic pain diseases [1] and inflammatory diseases [2]
The above two methods have their shortcomings in the quantification of macromolecules, which can be solved by liquid chromatography (LC)-high-resolution mass spectrometry (HRMS)
In the context of our study, we considered it necessary to adjust the UPLC conditions for analyzing the melittin in bee venom, especially in terms of the choice of additives for the mobile phase, because additives can aid in increasing separation [42], in addition to improving the response of high-level buffers

Summary

Introduction

Apis cerana Fabricius is a type of Asian honeybee whose venom has been widely applied as a traditional oriental medicine to treat some diseases such as chronic pain diseases [1] and inflammatory diseases [2]. The latest research has shown that bee venom exhibits potential therapeutic effects toward neurological diseases, such as amyotrophic lateral sclerosis (ALS) [3] and Parkinson’s Disease (PD) [4,5], in addition to peripheral neuropathy [6,7,8], cancer [9,10,11], and circulatory diseases [12]. Melittin is the main component of bee venom and exhibits many excellent biological activities such as antibacterial [14,15,16], antiviral [17,18], and anticancer properties [19,20,21].

Methods

Results

Discussion

Conclusion