In this protocol a minimum amount of 0.25% (w/w) of flavonoids and 0.50% (w/w) for phenolics in extracts of propolis extracts is established.
The authenticity of the extracts was established by phytochemical comparison against reference extracts prepared from authenticated E. arvense herbs with the traceability documents provided by each manufacturer and if dried raw herbs were available by genomic authentication.
The preparation of parasite extracts followed established protocols [21].
The criteria of cytotoxicity activity for the crude extracts, as established by the American National Cancer Institute NCI) is an IC50 < 30 μg/ml in the preliminary assay [ 34].
The criterion for cytotoxicity for the crude extracts, as established by the National Cancer Institute (NCI), is an IC 50 value lower than 30 μg/mL [ 67].
The 4 day suppressive test, which mainly evaluates the antimalarial activity of candidates on early infections, and Rane's test, which evaluates the curative capability of candidate extracts on established infections, are commonly used for antimalarial drug screening.
For the estimation of the in vitro cytotoxic potency of the investigated extracts, an established microtiter plate assay [ 9] was used with three human cancer cell lines: one lung cancer (A-427), one urinary bladder cancer (5637) and one breast cancer (MCF-7) line.
Additional efforts to improve this production process toward cGMP compliance include (a) developing methods for sterile filtration of the extract, (b) establishing extract process production reproducibility, (c) establishing the translational fidelity of OCFS produced proteins (Heibeck et al., manuscript in preparation), and (d) accurate cost-modeling of the reagents and other process variables.
The polyphenol compounds of olive leaf extract were established by using chromatographic analysis using GC-MS in center lab of Ain Shams University, Faculty of science.
Because the use of yeast extract in the complex medium was not economical, the production of AMVL in the optimal defined medium without yeast extract was established for a comparison.
