Triphala and Punarnava Show Complementary Antioxidant, Anti-Inflammatory Power in Lab Tests

Two of the most storied remedies in the Ayurvedic pharmacopoeia have just been put through a rigorous modern laboratory interrogation, and the results suggest that their ancient reputations rest on measurable biochemistry. In a study published in BMC Complementary Medicine and Therapies, researchers Shivani Makhijani and Deepak Khobragade of Datta Meghe College of Pharmacy, Datta Meghe Institute of Higher Education and Research in Wardha, India, compared the antioxidant and anti-inflammatory activities of hydroalcoholic extracts of Triphala and Punarnava, the latter derived from the plant Boerhavia diffusa. Their findings reveal a striking pharmacological division of labor: Triphala emerged as the stronger antioxidant, while Punarnava demonstrated superior anti-inflammatory effects in two established assays. The work, conducted entirely in vitro, offers a molecular rationale for why these botanicals have persisted in traditional practice for centuries, and hints at how they might be combined in future therapeutic strategies.

The choice of these two botanicals was anything but arbitrary. Triphala, whose name literally means three fruits, is a classical formulation combining the dried fruits of Amalaki (Emblica officinalis), Bibhitaki (Terminalia bellirica) and Haritaki (Terminalia chebula). It has long been prized in Ayurveda as a rejuvenating Rasayana, credited with benefits ranging from digestive support to eye health, and modern phytochemistry has attributed much of its activity to a rich arsenal of polyphenols, tannins, gallic acid and vitamin C. Punarnava, meanwhile, derives its name from the Sanskrit for renewing or reviving, and the creeping herb Boerhavia diffusa has been used traditionally to treat inflammation, liver disorders and urinary complaints. Despite this long history of parallel use, the two remedies had rarely been evaluated head to head under identical analytical conditions, a gap the Indian team set out to close.

A key methodological innovation of the study lies in how the plant material was extracted. Rather than relying on conventional maceration or Soxhlet techniques, which can be slow and thermally punishing, the researchers employed microwave-assisted extraction, or MAE, using a hydroalcoholic solvent mixture of 70 percent ethanol and 30 percent water. The extraction was performed at 200 watts and 50 degrees Celsius for just 18 minutes. This approach exploits the ability of microwave energy to heat solvent and plant cell moisture from within, rupturing cell walls rapidly and liberating bioactive compounds efficiently while minimizing degradation of heat-sensitive molecules such as phenolics and tannins. The ethanol water blend was chosen deliberately: water extracts highly polar constituents, ethanol captures moderately polar phenolics, and the combination maximizes the diversity of compounds recovered from both formulations.

With extracts in hand, the team assembled an unusually comprehensive battery of antioxidant tests, seven in total, each probing a different chemical facet of radical-scavenging capacity. The DPPH assay measures the ability of antioxidants to neutralize a stable synthetic nitrogen radical, providing a rapid readout of hydrogen-donating capacity. The ABTS assay extends this logic to both hydrophilic and lipophilic antioxidants by generating a green chromophore that decolorizes when reduced. The FRAP and CUPRAC assays assess reducing power, quantifying the capacity to convert ferric ions to ferrous ions and cupric ions to cuprous ions, respectively, which serves as a proxy for total antioxidant potential. Complementing these four, the researchers measured scavenging of nitric oxide, hydroxyl radical and superoxide radical, three reactive species with direct biological relevance, since these molecules are generated in living tissues during inflammation and contribute to oxidative damage of lipids, proteins and DNA.

Across the DPPH, ABTS, FRAP and CUPRAC assays, Triphala consistently outperformed Punarnava, achieving lower IC50 or EC50 values, meaning smaller concentrations of extract were needed to produce a half-maximal effect. This concentration-dependent superiority is chemically plausible: the three fruits of Triphala are famously dense in gallic acid, ellagic acid, chebulinic acid and ascorbic acid, compounds whose structures are optimized for electron donation and radical stabilization. The result positions Triphala as a broad-spectrum antioxidant capable of intercepting multiple classes of reactive oxygen and nitrogen species, the molecular vandals implicated in oxidative stress, a state now associated with aging, cardiovascular disease, neurodegeneration, diabetes and chronic inflammatory conditions.

The anti-inflammatory half of the investigation used two complementary assays that model different mechanisms of tissue protection. The protein denaturation assay examines whether an extract can prevent the structural unfolding of proteins, such as bovine serum albumin, under stress, since protein denaturation is thought to trigger autoimmune responses and inflammation in vivo, a mechanism implicated in rheumatic diseases. The proteinase inhibition assay, by contrast, tests the capacity to block proteolytic enzymes such as trypsin, which are released during inflammatory episodes and contribute to tissue destruction. Non-steroidal anti-inflammatory drugs, the clinical standard, are known to act partly through these mechanisms, making the assays a meaningful benchmark for botanical candidates.

Here the rankings flipped. Punarnava exhibited stronger anti-inflammatory activity than Triphala in both the protein denaturation and proteinase inhibition assays, again in a concentration-dependent fashion. The researchers interpret this as evidence that Boerhavia diffusa contains constituents particularly adept at stabilizing protein structure and restraining proteolytic cascades. The plant is known to harbor alkaloids such as punarnavine, along with flavonoids, lignans and ecdysteroids, any of which could underlie this protective behavior, although the present study did not attempt compound-level attribution. What matters from a pharmacological standpoint is the pattern: the two botanicals are not redundant but complementary, each excelling where the other is merely competent.

That complementarity is the study’s most intriguing implication. Oxidative stress and inflammation are deeply intertwined pathologies: reactive oxygen species activate inflammatory signaling pathways, including those involving tumor necrosis factor-alpha, and inflammatory cells in turn generate more free radicals, creating a self-amplifying loop. A therapeutic strategy that pairs a potent antioxidant with a potent anti-inflammatory agent could theoretically interrupt this loop at both ends. The authors suggest that Triphala and Punarnava, used together, could serve as natural sources for the development of new complementary therapeutic modalities for managing diseases and disorders in which both oxidative damage and inflammation play driving roles. Such conditions include, plausibly, metabolic syndrome, arthritis and ocular disorders, where both mechanisms converge.

Important caveats temper the enthusiasm, and the authors are careful to acknowledge them implicitly by framing the work as in vitro. Test-tube assays demonstrate chemical activity but say nothing about whether the active compounds survive digestion, reach target tissues in sufficient concentrations, or exert comparable effects in the complexity of a living organism. Bioavailability, metabolism, dosing and safety in humans all remain open questions that will require animal studies and, ultimately, controlled clinical trials. Nor did the study characterize the extracts’ full chemical fingerprints or isolate the specific molecules responsible for each activity. Nevertheless, by applying standardized, quantitative, concentration-dependent benchmarks to two venerable botanicals prepared with a modern extraction technology, the research provides a reproducible analytical foundation for that next phase. It transforms what has often been traditional assertion into testable hypothesis, and in doing so hands formulation scientists a provocative pairing: Triphala to quench the radicals, Punarnava to calm the inflammatory response, each doing what it demonstrably does best.

Subject of Research: Comparative in vitro evaluation of antioxidant and anti-inflammatory activities of Triphala and Punarnava extracts

Article Title: In vitro assessment of antioxidant and anti-inflammatory activities of Triphala and Punarnava: a comparative study

Article References: Makhijani, S., & Khobragade, D. (2026). In vitro assessment of antioxidant and anti-inflammatory activities of Triphala and Punarnava: a comparative study. BMC Complementary Medicine and Therapies. https://doi.org/10.1186/s12906-026-05580-3

Image Credits: AI Generated

DOI: 10.1186/s12906-026-05580-3

Keywords: Triphala, Punarnava, Boerhavia diffusa, antioxidant activity, anti-inflammatory activity, microwave-assisted extraction, DPPH, ABTS, FRAP, CUPRAC, Ayurveda, in vitro study