# LC-MS/MS Fingerprinting Analysis of Cyanotis arachnoidea Extracts: Process-Related Artifacts in Anabolic Food Supplements

## TL;DR

This paper identifies new ecdysteroids and process-related compounds in Cyanotis arachnoidea extracts, highlighting potential impacts on supplement quality and safety.

## Contribution

The study reports four new ecdysteroids and identifies autoxidation artifacts in processed extracts.

## Key findings

- Autoxidation of 20-hydroxyecdysone produces unknown ecdysteroids.
- 146 ecdysteroids were detected, with 20 identified using reference standards.
- Process-related artifacts were confirmed in commercial ecdysteroid extracts.

## Abstract

We report the isolation
and complete NMR characterization of four
new ecdysteroids and LC-MS/MS fingerprinting of ecdysteroids in two
native and two industrially processed Cyanotis arachnoidea extracts along with an autoxidized product mixture of 20-hydroxyecdysone
(20E). The autoxidation of 20-hydroxyecdysone leads to the formation
of various unknown or uncharacterized ecdysteroid compounds, which
can significantly impact the quality and efficacy of commercial ecdysteroid-containing
supplements, potentially affecting their regulatory status and consumer
safety. A total of 146 ecdysteroids were detected. Among these, 20
were identified using authentic and fully characterized reference
standards, including the newly reported compounds. The autoxidative
origin of many process-related artifacts was confirmed in the two
commercial ecdysteroid extracts. Considering the pharmacological versatility
of ecdysteroids and the major pharmacological differences between
these compounds and their autoxidized derivatives, our results are
of importance regarding the efficacy and safety of commercially available
ecdysteroid-containing food supplements.