Exploitation of HPLC Analytical Method for Simultaneous Determination of Six Principal Unsaturated Fatty Acids in Oviductus Ranae Based on Quantitative Analysis of Multi-Components by Single-Marker (QAMS).

Shihan Wang,Yuanshuai Gan,Hong Kan,Yongsheng Wang,Xinxin Mao

doi:10.3390/molecules26020479

Abstract

As one of the featured products in northeast China, Oviductus Ranae has been widely used as a nutritious food, which contains a variety of bioactive unsaturated fatty acids (UFAs). It is necessary to establish a scientific and reliable determination method of UFA contents in Oviductus Ranae. In this work, six principal UFAs in Oviductus Ranae, namely eicosapentaenoic acid (EPA), linolenic acid (ALA), docosahexaenoic acid (DHA), arachidonic acid (ARA), linoleic acid (LA) and oleic acid (OA), were identified using UPLC-MS/MS. The UFAs identified in Oviductus Ranae were further separated based on the optimized RP-HPLC conditions. Quantitative analysis of multi-components by single-marker (QAMS) method was implemented in content determination of EPA, ALA, DHA, ARA and OA, where LA was used as the internal standard. The experiments based on Taguchi design verified the robustness of the QAMS method on different HPLC instruments and chromatographic columns. The QAMS and external standard method (ESM) were used to calculate the UFA content of 15 batches of Oviductus Ranae samples from different regions. The relative error (r < 0.73%) and cosine coefficient showed that the two methods obtained similar contents, and the method validations met the requirements. The results showed that QAMS can comprehensively and effectively control the quality of UFAs in Oviductus Ranae which provides new ideas and solutions for studying the active components in Oviductus Ranae.

Highlights

IntroductionOviductus Ranae, derived from dried tubal product of Rana temporaria chensinensisDavid, has the effects of anti-fatigue, nourishing the lungs and improving immunobiological activity [1,2,3]
Oviductus Ranae, derived from dried tubal product of Rana temporaria chensinensisDavid, has the effects of anti-fatigue, nourishing the lungs and improving immunobiological activity [1,2,3]
In the twodimensional isoabsorption diagram of diode array detector (DAD) (Figure 1A), the red area that is approximately elliptical represents the compound with high absorbance

Summary

Introduction

Oviductus Ranae, derived from dried tubal product of Rana temporaria chensinensisDavid, has the effects of anti-fatigue, nourishing the lungs and improving immunobiological activity [1,2,3]. Oviductus Ranae, derived from dried tubal product of Rana temporaria chensinensis. As a precious natural product in northeast China, Oviductus Ranae has been used for nearly thousands of years and has developed into a series of nutrition, health food and traditional Chinese medicine. The content of UFA is considered to be closely related to the quality of Oviductus Ranae [15]. Due to the influence of breeding environment, production technology and preservation conditions [2], the content of UFA in Oviductus Ranae in different regions varies, resulting in the uneven quality of Oviductus Ranae in the market [15]. It is necessary to develop an economical and convenient method to monitor the content of UFA in Oviductus Ranae to ensure its quality

Methods

Results

Conclusion