LC-ESI-MS/MS analysis of phenolic compounds and in vitro cytotoxicity and apoptosis-inducing effects of Nonea pulmonarioides extracts.

The chemical composition of Nonea pulmonarioides extracts were investigated for the first time. The phytoconstituents of the methanol extracts were screened by using LC/MS-MS technique. The anticancer activity of the acetone and methanol extracts were measured against four cancer cell lines; MCF-7, PC3, HT-29, and U-87 MG. Thirty phenolic compounds were identified, rosmarinic (90.06 mg analyte/g extract) and fumaric acids (39.737 mg analyte/g extract) were major compounds of the studied species. Moreover, both methanol and acetone extracts were found to have strong anticancer activities. The acetone extract HT-29 (with IC50 of 10.17 ± 0.25 µg/mL) compared with standard cis-platin (with IC50 of 22.20 ± 0.72 µg/mL) with apoptotic mediated programmed cell death. These findings identified N. pulmonarioides as a potential species exhibiting anticancer properties. In conclusion, the compelling results show that the methanol extract contains possible bioactive compounds with anticancer properties that require isolation and further characterisation.

Chemical Profile, Antioxidant and Enzyme Inhibition Activities of Natural Saudi Sidr and Talh Honeys.

Honey is used since ancient time as a food and to cure many diseases. The present study investigated the chemical constituents, antioxidant and enzyme inhibition activities of natural Saudi Sidr (SH) and Talh (TH) honeys. Beside entire honey samples, ethyl acetate, ethanol and water extracts were prepared. The total polyphenolic content of SH, TH and their extracts was in the range of 2.86-7.21 and 3.80-17.33 mg gallic acid equivalents/g, respectively and the total flavonoids content was in the range of 0.05-1.17 and 0.18-2.38 mg rutin equivalents/g, respectively. Out of the 53 standards analyzed by HPLC, 27 compounds were detected with highest number of compounds identified in the ethyl acetate extract of TH (45 %, 24/53) and SH (26 %, 14/53), respectively. Quinic acid was dominant compound identified in all honey samples with the highest content determined in TH ethanol extract (4454 µg/g). The majority of tested samples possessed considerable anti-radicals and reducing ions capacity with the ethyl acetate extract from TH exerted significantly (p<0.05) the highest activity. All honey samples did not show chelating iron metal property. Honey samples revealed variable enzyme inhibition activity with TH (entire and/or ethyl acetate extract) showed significantly (p<0.05) the highest acetylcholinesterase, butyrylcholinesterase, tyrosinase and α-amylase inhibition activity. In conclusion, ethyl acetate is the best solvent for extraction of bioactive molecules from the two honey types. Moreover, the dark-colored TH contained the highest number of molecules and consequently exerted the best antioxidant and enzyme inhibition activities in most assays.

A comprehensive LC-MS/MS method validation for the quantitative investigation of 37 fingerprint phytochemicals in Achillea species: A detailed examination of A. coarctata and A. monocephala.

The current study aims to optimize and validate a comprehensive LC-MS/MS method for the quantification of 37 phytochemicals (15 phenolic acids, 17 flavonoids, 3 non-phenolic organic acids, 1 phenolic aldehyde and 1 benzopyrene) in Achillea species. Though Achillea species were chosen as real life samples, the current method is applicable to a wide range of plant species. The developed method was fully validated in terms of linearity, accuracy (recovery), inter-day and intra-day precision (repeatability), limits of detection and quantification (LOD/LOQ) and relative standard uncertainty (U% at 95% confidence level (k = 2)). Reversed-phase ultrahigh performance liquid chromatography was optimized to achive optimum separation for 37 phytochemical compounds and to overcome the suppression effects. MS detection was performed using a triple quadrupole mass spectrometer and negative or positive ionization modes were optimized for each analyte. Multiple reaction monitoring (MRM) was used to quantify the analytes, related molecular ions and transition ions were optimized. Phytochemical screening of ethanol and methanol-chloroform extracts of root and aerial parts of A. coarctata and A. monocephala were performed by using the developed and validated LC-MS/MS method. Root and aerial parts of both species have considerable amounts of certain phenolic-nonphenolic acids (quinic, malic, fumaric, chlorogenic and vanillic acids) and flavonoids (rutin, hesperidin, isoquercitrin, apigetrin, luteolin, apigenin). Additionally, total phenolic and flavonoid amounts, antioxidant (DPPH free radical scavenging assay, ABTS radical cation decolorization assay, ß-carotene lipid peroxidation test system and CUPRAC cupper reduction capacity methods), anticholinesterase, tyrosinase, urease inhibition and cytotoxic activities (on HeLa (Human Cervical Carcinoma Cell Line) of A. coarctata and A. monocephala were also investigated. It has been determined that the studied Achillea species, that are rich in total phenolic-flavonoid and chlorogenic acid contents, have high antioxidant and cytotoxic potential at the same time. According to the results of LC-MS/MS, antioxidant and cytotoxic activity studies, after detailed chemical investigation and toxicity studies on these species, A. coarctata and A. monocephala may be promoted as promising sources of natural agents and used for the development of nutraceuticals or functional food ingredients in future.

Comparison of reproductive toxicity caused by cisplatin and novel platinum-N-heterocyclic carbene complex in male rats.

Cisplatin and other platinum complexes are important chemotherapeutic agents and useful in the treatment for several cancers such as prostate, ovarian and testis. However, severe side effects including reproductive toxicity of cisplatin and other platinum complex cause limitations in their clinical usage. In this context, we aimed to compare the damage in testis caused by cisplatin and a novel platinum-N-heterocyclic carbene complex (Pt-NHC). To this end, 35 Sprague-Dawley rats were divided randomly into five equal groups (n = 7 in each group). Cisplatin and Pt-NHC were intraperitoneally administered as a single dose of 5 mg/kg or 10 mg/kg, and the rats were then killed 10 days after this treatment. The testicular tissues and serum samples were taken from all rats for the determination of reproductive toxicity. The results showed that cisplatin and Pt-NHC caused toxicity on the reproductive system via increased oxidative and histological damage, decreased serum testosterone levels and negatively altered sperm characteristics in a dose-dependent manner (p < 0.05). At the same dose levels, cisplatin generally caused lower toxicity on the reproductive system compared with Pt-NHC. In conclusion, these results suggest that Pt-NHC has more toxic effects on the male reproductive system than cisplatin, and in terms of clinical usage, Pt-NHC may be unsafe compared with cisplatin.

The determination of oxidative damage in heart tissue of rats caused by ruthenium(II) and gold(I) N-heterocyclic carbene complexes.

In the present study, we aimed to determine the oxidative damage in rat heart tissue induced by ruthenium(II)-NHC (Ru) and gold(I)-NHC (Au) complexes which have anticarcinogenic effects and not used clinically yet. For this purpose, 35 Sprague-Dawley rats were randomly divided into 5 equal groups. In the control group, rats treated with saline, Ru and Au complexes were intraperitoneally given high (10 mg/kg) and low (5 mg/kg) doses as only one administration. The animals were killed, and heart tissues were taken on day 10 of the drug administration for the determination of the biochemical parameters (malondialdehyde, superoxide dismutase, reduced glutathione and catalase levels). It was determined that both Ru and Au complexes treatment significantly caused oxidative damage compared to the control group. Additionally, it was shown that Au treatment caused more adverse effects than Ru treatment. Also, it was clearly found that the occurred effects were generally determined in a dose-dependent manner. In conclusion, when these compounds synthesized for the treatment of cancer were used, they caused oxidative damage in heart tissue. However, Ru complex could be preferred for cancer treatment in terms of user safety.