New Method Developed for Quantifying Natural Bioactive Compounds in Serum

The properties in natural bioactive compounds (NBCs) can be used to treat various illnesses such as diabetes and Alzheimer’s disease (1). There are several NBCs that are often used in clinical applications, including polyphenols, resveratrol, proanthocyanidins, alkaloids, and berberine. NBCs also have played a role in several traditional Chinese medicines.

Studies have examined how NBCs in traditional Chinese medicines can be used to treat various respiratory illnesses, such as COPD, asthma, pneumonia, tuberculosis, and lung cancer (2). In a recent study published in Analytical and Bioanalytical Chemistry, lead researchers Yong-hui Mao and Cai Tie from Beijing, China, and their team sought to address challenges associated with quantifying NBCs that are found in Tripterygium wilfordii Hook f., commonly known as Lei-Gong-Teng or thunder god vine (3).

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In thunder god vine, the NBCs in this traditional Chinese medicine are integral to treating many human afflictions such as rheumatoid arthritis (RA), nephrotic syndrome (NS), and autoimmune hepatitis (AIH). However, because of matrix effect and real standard deviation (RSD), accurately quantifying NBCs in serum have encountered many obstacles (3).

To overcome these obstacles, the research team developed a new quantitation method utilizing ¹⁸O labeling combined with off-line solid-phase extraction (SPE) (3). By contrasting the recoveries and matrix effects of various separation methods, the researchers identified the optimal technique. Additionally, they optimized conditions for SPE loading and washing, crucial steps in the quantitation process (3).

An isotopic internal standard was prepared through the ¹⁶O/¹⁸O exchanging reaction, effectively eliminating matrix effects. The method demonstrated high accuracy and precision, meeting the requirements for method validation with a recovery rate close to 60%. The relative standard deviation (RSD) for high-concentration samples was a mere 2%, with a limit of detection (LOD) as low as 1 ng/mL.

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To evaluate the method's clinical applicability, serum samples from 10 patients with diabetes nephropathy were analyzed for demethylzeylasteral concentration. The results showed a correlation between therapeutic drug monitoring (TDM) and decreased urinary protein, indicating the potential utility of this method in clinical settings.

This innovative approach not only addresses the challenges of quantifying low-abundance and difficult-to-quantify NBCs but also holds promise for advancing the study of drug metabolism in vivo. With its implications for improved clinical treatment strategies, this research marks a significant advancement in the field of analytical chemistry and pharmaceutical science.

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References

(1) Huang, J.; Huang, N.; Mao, Q.; et al. Natural Bioactive Compounds in Alzheimer's Disease: From the Perspective of Type 3 Diabetes Mellitus. Front. Aging Neurosci. 2023, 15, 1130253. DOI: 10.3389/fnagi.2023.1130253

(2) Sindhu, R. K.; Sofat, M.; Kaur, H.; et al. Nanodeliovery based Chinese Medicine's Bioactive Compounds for Treatment of Respiratory Disorders. Pharm. Res. – Modern Chinese Medicine 2022, 5, 100180. DOI: 10.1016/j.prmcm.2022.100180

(3) Wang, Y.; Zhao, B.; Dang, X.; et al. A Novel SPE-LC-MRM Strategy for Serum Demethylzeylasteral Quantitation Developed with an ¹⁸O-labeled Internal Standard. Anal. Bioanal. Chem. 2024, 416, 467–474.