Correlation of In Vivo and In Vitro Assay Results for Assessment of Free Radical Scavenging Activity of Green Tea Nutraceuticals

Green tea (GT)‐derived catechins; epigallocatechin gallate (EGCG) in particular are commonly used nutraceuticals for their free‐radical scavenging activity (FRSA). The influence of photodegradation on the protective power of GT nutracenticals against oxidative stress was thoroughly explored. Photodegradation of GT extracts was carried out and monitored using orthogonal stability‐indicating testing protocol; in vitro and in vivo assays. Total polyphenol content (TPC) and FRSA were determined spectrophotometrically while EGCG was selectively monitored using SPE‐HPLC. In vivo assessment of photodegraded samples was investigated via measuring a number of biomarkers for hepatic oxidative stress and apoptosis (caspase‐3, inducible nitric oxide synthase, nitric oxide, mitogen‐activated protein kinase, glutathione, thiobarbituric acid reactive substances, nuclear factor kappa beta, and nuclear factor erythroid 2‐related factor) as well as liver damage (alanine transaminase and aspartate transaminase) in serum of rats previously subjected to oxidative stress. Results showed complete degradation of EGCG in photodegraded green tea samples with no correlation with either TPC or FRSA. On the other hand, in vivo assay results revealed not only loss of activity but formation of harmful pro‐oxidants. Photostability was found crucial for the protective effect of GT extract against lead acetate insult. Results confirmed that careful design of quality control protocols requires correlation of chemical assays to bioassays to verify efficacy, stability, and most importantly safety of nutraceuticals. Practical Application In this study, an orthogonal analysis protocol was designed to evaluate the photostability of a model green tea nutraceutical formulation. Lack of agreement between chemical assay results required correlation to in vivo activity in animal model. Results indicated that photodegraded green tea samples were not only ineffective but also harmful to biological systems. Photodegradation products exerted prooxidant effect rather than the beneficial antioxidant effect. Results of the study confirmed that inappropriate selection of analysis techniques and lack of correlation between in vitro and in vivo assay results might lead to elusive conclusions about the quality of nutraceuticals.