Herbal extracts are gaining widespread acceptance as ingredients in cosmetic and health products. Moringa oleifera Lam. is an interesting candidate as it has multiple biological properties. However, methods for quality control of the use of M. oleifera are still required. This study proposes the use of isothiocyanate compounds and astragalin as chemical markers to standardize M. oleifera extracts. Both of these compounds are known to have anti-inflammation properties, and astragalin is also reported to have antioxidant properties. The HPLC-based cyclocondensation method was further developed and validated specifically for quantitative analysis of the isothiocyanate compounds in the M. oleifera extracts, and the concentration of astragalin in the M. oleifera extracts was determined by the HPLC method. Both methods gave acceptable linearity, accuracy, and precision with the limit of detection of 0.13 µg/ml for total isothiocyanate and 0.10 µg/ml for astragalin. The uses of the methods were demonstrated in the analyses of the components in the extracts taken variously from the leaves, immature seeds, mature seeds, petioles, and pods of the M. oleifera tree. Our results suggest that the methods developed could be used in the quality control of M. oleifera extracts to develop cosmetic and natural health products.

Brahmi essence, developed from Bacopa monnieri (L.) Wettst. standardized extract and mulberry juice, was proven to improve the memory speed of healthy participants aged 55–80 years old, following a 12-week dietary program. However, the metabolites have not yet been reported. Our objective was to characterize the altered metabolites in the plasma, urine, and feces of healthy volunteers after consumption of Brahmi essence for 12 weeks, using the LC-MS metabolomics approach. The altered metabolites were selected from OPLS-DA S-plots; 15 metabolites in the plasma, 7 in the urine, and 17 in the feces samples were tentatively identified by comparison with an online database and literature. The metabolites in the plasma samples were in the classes of amino acids, acylcarnitine, and phospholipids. Benzeneactamide-4-O-sulphate and 3-hydroxyhippuric acid were found in urine samples. The metabolites in the class of amino acids, together with jujubogenin and pseudojujubogenin, were identified in the fecal samples. The aminoacyl-tRNA, aromatic amino acids, and branched-chain amino acid biosynthetic pathways were mainly related to the identified metabolites in all three samples. It could be implied that those metabolites and their pathways might be linked with the effect of Brahmi essence on memory speed.

Eulophia macrobulbon (E.C.Parish & Rchb.f.) Hook.f. contains a natural PDE5A1 inhibitor, phenanthrene, 1-(4'-hydroxybenzyl)-4,8- dimethoxyphenanthrene-2,7-diol (HDP), a potential agent for the treatment of erectile dysfunction. The aim of this study was to improve the extraction efficiency of HDP from E. macrobulbon by using a more environmentally friendly extraction method, subcritical liquid dimethyl ether extraction (sDME), instead of classical solvent extraction (CSE) and ultrasound-assisted extraction (UAE). The efficiency and quality of the extracts obtained were evaluated using the following criteria: %process yield; solvent amount; extraction time; temperature; %HDP content by LC–MS, bioactivity as inhibition of phosphodiesterase-5A1 (PDE5A1) by radio-enzymatic assay; and chemical profiles by LC-QTOF-MS. sDME provided the highest content of HDP in the extract at 4.47%, much higher than the use of ethanol (0.4–0.5%), ethyl acetate (1.2–1.7%), or dichloromethane (0.7–1.4%). The process yield for sDME (1.5–2.7%) was similar to or lower than the other solvents (0.9–17%), but as long as the process yield is not prohibitively low, the concentration is a more important measure for clinical use. The optimal conditions for sDME extraction were: Extraction time, 40 min; 200% water as co-solvent; sample-to-solvent ratio of 1:8; temperature, 35 °C. Phenanthrene aglycone and glycoside derivatives were the major constituents of the sDME extracts and lesser amounts of phenolic compounds and sugars. The inhibition of PDE5A1 by sDME (IC50 0.67 ± 0.22 µg/ml) was tenfold more potent than ethanolic extract and other extraction methods, suggesting a high probability of clinical efficacy. Thus, sDME was a more efficient, faster, solvent-saving and environmentally friendly extraction method and more selective for phenanthrene when extracted from E. macrobulbon.

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Eulophia macrobulbon (E.C.Parish & Rchb.f.) Hook.f. contains a natural PDE5A1 inhibitor, the phenanthrene, 1-(4'-hydroxybenzyl)-4,8- dimethoxyphenanthrene-2,7-diol (HDP) a potential treatment for erectile dysfunction. This investigation aimed to improve extraction efficiency of HDP from E. macrobulbon by using greener extraction methodology, subcritical fluid dimethyl ether extraction (sDME) rather than classical solvent extraction (CSE) and ultrasound-assisted extraction (UAE). The efficiency and quality of obtained extracts were evaluated by: %process yield; solvent amount; extraction period; temperature; %HDP content by LC-MS assay, bioactivity as inhibition of phosphodiesterase-5A1 (PDE5A1) by radio-enzymatic assay; and chemical profiles by LC-QTOF-MS analysis. sDME yielded the highest content of HDP in the extract at 4.47%, much higher than using ethanol (0.4-0.5%), ethyl acetate (1.2-1.7%), or dichloromethane (0.7-1.4%). Process yield for sDME (1.5-2.7%) was similar or less than that observed with other solvents (0.9-17%), but providing that process yield is not prohibitively low, concentration is a more important metric for clinical application. Optimal sDME extraction conditions were: extraction period, 40 mins; 200% water as a cosolvent; sample-to-solvent ratio of 1:8; temperature, 35°C. Phenanthrene aglycone and glycoside derivatives were major constituents in sDME extracts and lesser amounts of phenolic compounds and sugars. Inhibition of PDE5A1 by sDME (IC50 0.67±0.22 µg/mL) was 10-fold more potent than the ethanolic extract and other extraction methods, suggesting a high likelihood of clinical efficacy. Thus, sDME was more efficient, faster, solvent-sparing, greener extraction methodology and more selective for phenanthrene when extracted from E. macrobulbon.