Janatová, M.: Determination of amygdalin content in trade stone fruits and its biological activity in cultured cancer cells. Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2015, pp.74 Stone fruits from tribe Amygdaleae of Rosaceae family are known for their antioxidant activity and amount of nutrients and vitamins. Their seeds are connected with content of cyanogenic glycoside amygdalin and their possible effect on inhibition of cancer cells growing. The aim of study is to quantitatively analyze amount of amygdalin in extracts of trade stone fruit seeds (apricots, almonds, plums, cherries, sour cherries, peaches) and one dietary supplement B17 APRICARC®. Kernels were extracted in methanol by Soxhlet extractor and subsequently analyzed by TLC and by reverse phase HPLC with UV detection on 214-215 nm wavelength. The highest concentration of amygdalin was found in apricot seeds during their ripeness with the median value 79.59 mg/g of seeds. On the other hand, in sweet almonds and apricots from Pakistan was not detected any amount. The enzymatic hydrolysis by β-glucosidase and degradation of amygdalin was studied in vitro and analyzed by reverse- phase High performance liquid chromatography (rp-HPLC). Anticancer...

1. Colorimetric Assay for Cyanide and Cyanogenic Glycoside Using Polysorbate 40-Stabilized Gold Nanoparticles. This study described a simple and homogeneous method for the selective and sensitive detection of cyanide and endogenous biological cyanide using polysorbate 40-stabilized gold nanoparticles (PS 40-AuNPs). Neutral PS 40 molecules enable citrate-capped AuNPs to stabilize in a high-salinity solution. The addition of cyanide to a solution of PS 40-AuNPs resulted in the formation of AuCN(s) on the NP surface and Au(CN)2¡V in an aqueous solution. The removal of PS 40 molecules from the NP surface rendered the AuNPs unstable in a high-salinity solution, leading to NP aggregation. The formation of AuCN(s) and Au(CN)2¡V was demonstrated by means of surface-assisted laser desorption/ionization time of flight mass spectrometry and inductively coupled plasma mass spectroscopy, respectively. PS 40-AuNPs were capable of selectively detecting cyanide at concentrations as low as 500 nM. Additionally, the minimum detectable concentration of linamarin (cyanogenic glycoside) was measured to be 1 uM using PS 40-AuNPs. This probe was successfully applied to the determination of cyanide in tap water, the monitor of cyanide removal during food processing, and the quantification of linamarin in cassava root. 2. Colorimetric detoxification and monitored cyanogenic glycoside in plants/fruit using polysorbate 40-stabilized gold nanoparticles. Developing rapid, highly sensitive, and selective detection/inhibition of cyanide/cyanogenic glycoside from plants and foods is extremely essential for human life safety. Here we report a strategy for the colormetric visualization of cyanogenic glycoside using polysorbate 40 stabilized gold nanoparticle (PS 40-AuNPs). Two cyanogenic glycosides (amygdalin and linamarin) were chosen to determine the efficiency of acid hydrolysis. According to US Department and Health and Human Services standard cyanide antidote kit, sodium thiosulfate and hydroxocobalamin (vitamin B12a) seems to be an appropriate antidote for treatment cyanide poisoning victims. The addition of thiosulfate/vitamin B12a to a solution of cyanide/cyanogenic glycosides resulted in the formation of thiocyanate/vitamin B12 in an aqueous solution, which couldn¡¦t etch PS 40-AuNPs and inhibit the aggregation of PS 40-AuNPs in a high-salt solution. The inhibition/detoxification efficiency (IC50) of thiosulfate and vitamin B12a were studied for treatment of cyanide and hydrolyzed cyanogenic glycoside. This probe was also used to monitor the removal of cyanide, estimated the concentration of cyanide and detoxification of cyanide by thiosulfate in plants/fruit sample.

Magister Scientiae (Medical Bioscience) - MSc(MBS) / Amygdalin, a controversial anti-cancer agent, is a cyanogenic glycoside plant compound found in apricot and peach kernels. Both amygdalin and its patented form, Laetrile®, have been promoted and sold as "vitamin B-17", although neither compound is a vitamin. No consensus on the efficacy of amygdalin regarding the treatment of different cancers has been reached. Cancer is now the third leading cause of death worldwide. More than 7.6 million deaths were estimated to have occurred in 2007 and by 2030 it is projected to increase to 17 million cancer deaths per year. Cancers of the lung, breast, colon/rectum, liver and prostate are no longer largely confined to Western industrialized countries but are among the most common cancers worldwide (Thun et al. 2010). In South Africa it is estimated that one in every four males and one in every five females will be affected by a cancer diagnosis in their lifetime. The most common cancers in males are prostrate, lung, oesophagus, bladder and colorectal and in females they are cervix, breast, colorectal, oesophagus and lung (Haggar & Boushey 2009). Colon cancer is one of the most prevalent cancers worldwide, especially in western societies and is nutrition dependent (Klenow et al. 2009). It is one of the leading causes of death in both men and women in industrialised western countries. Colon cancer development involves both hereditary factors and lifestyle factors which include absence of physical exercise, unbalanced nutrition and long term smoking (Forman et al. 2004; Heavey et al. 2004). Colon cancer is traditionally treated by the resection of the colon, chemotherapy, radium therapy, and pharmaceutical hormonal drugs (Willson et al. 1987; Padussis et al. 2004)). Epidemiological studies supports evidence that colon cancer is preventable by adjusting the diet (Forman et al. 2004) and a protective effect is attributable to polyphenols and foods such as fruits and vegetables (Araújo et al. 2011). It was reported by Ruan et al. (2006) that the addition of Chinese Herbal Medicine in conjunction with chemotherapy notonly raised the efficacy of the chemotherapeutic drug, but also reduced the toxic side-effects. The aim of this research was to carry out a comparative in vitro study of the anti-tumour effect of the Chinese , South African and Turkish apricot (Xing ren / Armeniacea Semen) and Chinese and South African peach (Tao ren / Persica Semen) kernel extracts on the HT-29 colon cancer cell line.All the extracts significantly reduced cell viability and inhibited proliferation in the HT-29 cancer cells after 24 hours with the lipophilic and total fractions of CAK being the most effective. After 72 hours, it is clear that the inhibitory effects have been abolished and replaced by a stimulatory effect as the cell viability is higher in the treated cultures than the untreated controls. Results show that the total and the hydrophilic fractions of all the kernels increased cell viability more than the lipophilic fractions. It cannot be said with certainty that it was the amygdalin metabolite cyanide that affected the cell viability or induced apoptosis on its own. If hydrolysis of amygdalin indeed happened and cyanide was produced, it would affect the cells by shutting down aerobic respiration. Since cancer cells have more β- glucosidases and less rhodanese than normal cells, it is a possibility that the HT-29 cancer cells had some rhodanese to convert cyanide into a relatively harmless compound thiocyanate. It could be that in vitro this conversion, in light of the low enzyme levels in the HT-29 cancer cells, happened slowly and that the effect was only seen after 48 hour. However, this does not explain the overall inhibition even by the lipophilic fractions that should not contain any amygdalin or the eventual stimulatory effect, observed from 48 hour onwards.The S phase block observed, was mostly seen after 24 hour exposure to organic extractions, with the SAK showing 86% of cells in the S phase in contrast to the aqueous extractions which only slightly increased the S phase fraction. This could indicate that synergistic and/or additive effects between polyphenolic compounds may also be responsible for the reduction of cell viability, proliferation and apoptosis. All the kernels and the various fractions affected cell viability and to an extent cell cycle progression, but more studies is needed to establish the most effective kernel and specific fraction or signature active component. Inhibition of cell viability and proliferation and the induction of apoptosis could be an important preventive approach in chemoprevention. Understanding how dietary components regulate proliferation and cell survival could play a critical role in development of new enriched agents that can prevent and treat cancer with reduced risk of toxicity.