Chemical Impossibility in Phytochemical Quantification (Table 3): You report the Total Flavonoid Content (TFC) as 65.56 mg QE/g, which is over four times higher than the Total Phenolic Content (TPC) at 15.50 mg GAE/g. Since flavonoids are a chemical subclass of phenolics, TPC must inherently be greater than or equal to TFC. Please clarify if this is a unit conversion error, swapped data entries, or a fundamental flaw in your Folin-Ciocalteu/AlCl₃ assay calibration.
Statistically Invalid LD₅₀ Calculation (Table 7): In the brine shrimp lethality assay, your highest tested concentration (1000 µg/mL) resulted in exactly 50% mortality (10 dead, 10 survive). Probit analysis for LD₅₀ calculation statistically requires the mortality range to bracket the 50% threshold (i.e., the highest concentration must yield significantly >50% mortality to allow valid extrapolation). Calculating an LD₅₀ of 882.97 µg/mL from a maximum mortality of exactly 50% is mathematically unreliable. Please justify this statistical approach or provide the raw data showing a proper mortality gradient.
Anomalously “Perfect” MBC/MIC Ratios (Table 6): The MBC/MIC ratio is reported as exactly 2.0 for all four bacterial strains and exactly 4.0 for the fungal strain. In empirical biological replicates, it is highly improbable to observe such perfect integer multiples across diverse microorganisms. Please provide the raw colony-forming unit (CFU) counting data to demonstrate that these are actual empirical results, rather than assumed theoretical two-fold serial dilution steps.
Implausible Crude Extract Antioxidant Potency (Fig. 3 & Discussion): You claim the crude ethanolic extract has a DPPH IC₅₀ of 6.09 µg/mL, which is nearly three times more potent than the pure synthetic standard, propyl gallate (17.39 µg/mL). It is chemically implausible for a crude extract containing significant inactive biomass (e.g., sugars, pigments) to exhibit higher specific antioxidant potency per milligram than a pure compound. Please provide the raw standard curve data and exact sample weighing records to rule out severe dilution or calculation errors.
