HPLC-PDA Method for Quantification of Bioactive Compounds in Crude Extract and Fractions of Aucklandia costus Falc. and Cytotoxicity Studies against Cancer Cells.

Aucklandia costus Falc. (Synonym: Saussurea costus (Falc.) Lipsch.) is a perennial herb of the family Asteraceae. The dried rhizome is an essential herb in the traditional systems of medicine in India, China and Tibet. The important pharmacological activities reported for Aucklandia costus are anticancer, hepatoprotective, antiulcer, antimicrobial, antiparasitic, antioxidant, anti-inflammatory and anti-fatigue activities. The objective of this study was the isolation and quantification of four marker compounds in the crude extract and different fractions of A. costus and the evaluation of the anticancer activity of the crude extract and its different fractions. The four marker compounds isolated from A. costus include dehydrocostus lactone, costunolide, syringin and 5-hydroxymethyl-2-furaldehyde. These four compounds were used as standard compounds for quantification. The chromatographic data showed good resolution and excellent linearity (r2 ˃ 0.993). The validation parameters, such as inter- and intraday precision (RSD < 1.96%) and analyte recovery (97.52-110.20%; RSD < 2.00%),revealed the high sensitivity and reliability of the developed HPLC method. The compounds dehydrocostus lactone and costunolide were concentrated in the hexane fraction (222.08 and 65.07 µg/mg, respectively) and chloroform fraction (99.02 and 30.21 µg/mg, respectively), while the n-butanol fraction is a rich source of syringin (37.91 µg/mg) and 5-hydroxymethyl-2-furaldehyde (7.94 µg/mg). Further, the SRB assay was performed for the evaluation of anticancer activity using lung, colon, breast and prostate cancer cell lines. The hexane and chloroform fractions show excellent IC50 values of 3.37 ± 0.14 and 7.527 ± 0.18 µg/mL, respectively, against the prostate cancer cell line (PC-3).

Glutaminase inhibitory activity of umbelliferone isolated from kabosu (Citrus sphaerocarpa Hort. ex Tanaka).

Kabosu (Citrus sphaerocarpa Hort. ex Tanaka) fruits have pleasant and fresh odors and have been used as raw materials for vinegar, seasonings, jams, marmalades and juices in Japan. The n-butanol extracts from kabosu fruits were prepared and a component in the extract was purified by column chromatography and HPLC to afford compounds 1-3. Three compounds, 5-(hydroxymethyl)-2-furaldehyde (1), umbelliferone (2) and oxypeucedanin hydrate (3), have been isolated from kabosu, and the structures of compounds 1-3 were elucidated by 1 D and 2 D NMR as well as EI-MS. Compound 2 exhibited potent glutaminase inhibitory activity with an IC50 value of 1.33 mM. This is the first report on glutaminase inhibitory activity of 2 and the isolation of three compounds 1-3 from kabosu fruits.

Antioxidative Capacity of and Contaminant Concentrations in Processed Plum Products Consumed in Romania.

The bioactive compounds (total phenols, anthocyanins, and flavonoids) and antioxidant activity of plum-based products (prunes, jams, and marmalade) were evaluated based on spectrophotometric assays and acrylamide and 5-hydroxymethyl-2-furaldehyde concentrations to determine the quality and safety of these products in the Romanian marketplace. The total polyphenol concentrations in prunes, marmalade, and jams were 0.93 to 5.63 g of gallic acid equivalent per kg of dry matter, and the antioxidant activity was 3.0 to 17.2 mmol/kg. The concentrations of potentially harmful acrylamide in the plum products ranged from the limit of quantitation (<20 µg/kg) to 37.44 µg/kg, and the concentration was higher in prunes (26.66 to 163.72 µg/kg) than in jams. The 5-hydroxymethyl-2-furaldehyde concentrations in plum products were also variable, at 36 to 2,149 mg/kg. These data revealed a very low concentration of acrylamide in thermally processed plum products, and the results for total polyphenols and the antioxidant activity of prunes and plum jams may be relevant for evaluating these products as important sources of bioactive compounds.

Inhibitory mechanism of quercetin against the formation of 5-(hydroxymethyl)-2-furaldehyde in buckwheat flour bread by ultra-performance liquid chromatography coupled with high-resolution tandem mass spectrometry.

Ultra-performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UPLC-HRMS/MS) was employed to investigate the inhibitory mechanism of quercetin against the formation of 5-(hydroxymethyl)-2-furaldehyde (HMF) in buckwheat flour bread. The HMF and HMF precursors (3,4-dideoxyglucosone-3-ene (3,4-DGE), 3-deoxyglucosone (3-DG), or fructofuranosyl cation dehydration products (FCDPs)) adducts of quercetin were detected in buckwheat flour bread, with the trapping of these compounds by quercetin to form corresponding adducts with HMF or its precursors in 1:1, 1:2, 1:3, and 1:4 ratios (where "1" refers to quercetin in all cases). The structures of these adducts were elucidated by UPLC-HRMS/MS. Carbohydrate module labeling (CAMOLA) techniques and the labelled quercetin in model reactions were utilized to further confirm the inhibitory mechanism. Effects of baking temperature and time on the HMF inhibition rate were investigated in wheat flour bread, and a maximum inhibition rate of 86.0% was obtained with the baking of wheat flour bread (with the added quercetin concentration of 1.90mg/g) at 160°C for 30min.

A randomized, double-blind, placebo-controlled trial of a traditional herbal formula, Yukmijihwang-tang in elderly subjects with xerostomia.

ETHNOPHARMACOLOGICAL RELEVANCE: Yukmijihwang-tang (YMJ) is a typical herbal formula to treat Yin-deficiency (YD) syndrome by enriching the fluid-humor of the body. YMJ has been used to treat dry mouth symptoms for hundreds of years in traditional East Asian medicine. Xerostomia, a subjective oral dryness, is common in the elderly and results in impaired quality of life. Many conventional treatments for xerostomia provide only temporary symptom relief, and have side effects. The aim of this study is to investigate the efficacy and safety of YMJ for the treatment of xerostomia in the elderly. METHODS: This study was designed as a randomized, placebo-controlled, double-blinded, two center trial. Ninety-six subjects aged 60-80 years who had experienced xerostomia for at least 3 months and presented with score>40 on the visual analog scale (VAS) for subjective oral dryness were recruited and randomly allocated to YMJ and placebo groups. YMJ or placebo was administered to each group for 8 weeks (3g of YMJ or placebo, three times per day). The primary outcome was change of VAS for xerostomia from 0 to 8 weeks. RESULTS: VAS for xerostomia was decreased by 22.04±22.76 in the YMJ group and 23.58±23.04 in the placebo group. YMJ had no effect on xerostomia. However, participants with BMIs lower than 29.37kg/m(2) showed improvement of xerostomia after 8 weeks of treatment with YMJ compared to placebo. In addition, YMJ improved oral moisture, which is associated with subjective oral dryness in the YMJ group, and the relationship between VAS for xerostomia and YD was significant. CONCLUSION: A trend was observed in which YMJ improved oral moisture status and subjective oral dryness in elderly subjects with lower BMI and greater tendency toward YD.