The composition of potentially bioactive triterpenoid glycosides in red raspberry is influenced by tissue, extraction procedure and genotype.

The beneficial effects of consumption of berry fruits on a range of chronic diseases has been attributed (at least in part) to the presence of unique phytochemicals. Recently, we identified novel ursolic acid-based triterpenoid glycosides (TTPNs) in raspberry fruit and demonstrated their survival in human ileal fluids after feeding which confirmed their colon-availability in vivo. In this paper, in vitro digestion studies demonstrated that certain TTPNs were stable under gastrointestinal conditions and confirmed that these components may have been responsible for bioactivity noted in previous studies. Sequential extractions of raspberry puree, isolated seeds and unseeded puree showed that certain TTPN components (e.g. peak T1 m/z 679, and T2 m/z 1358) had different extractabilities in water/solvent mixes and were differentially associated with the seeds. Purified seed TTPNs (mainly T1 and T2) were shown to be anti-genotoxic in HT29 and CCD841 cell based in vitro colonocyte models. Further work confirmed that the seeds contained a wider range of TTPN-like components which were also differentially extractable in water/solvent mixes. This differential extractability could influence the TTPN composition and potential bioactivity of the extracts. There was considerable variation in total content of TTPNs (∼3-fold) and TTPN composition across 13 Rubus genotypes. Thus, TTPNs are likely to be present in raspberry juices and common extracts used for bioactivity studies and substantial variation exists in both content and composition due to genetics, tissue source or extraction conditions, which may all affect observed bioactivity.

[1]  A. Törrönen,et al.  Influence of domestic processing and storage on flavonol contents in berries. , 2000, Journal of agricultural and food chemistry.

[2]  T. Nohara,et al.  Triterpenoids from the fruits and leaves of the blackberry (Rubus allegheniensis) and their inhibitory activities on foam cell formation in human monocyte-derived macrophage , 2014, Natural product research.

[3]  C. Finn,et al.  Antioxidant properties of domesticated and wild Rubus species , 2000 .

[4]  A. Rodriguez-Mateos,et al.  Dietary (poly)phenolics in human health: structures, bioavailability, and evidence of protective effects against chronic diseases. , 2013, Antioxidants & redox signaling.

[5]  Eric J Hanson,et al.  Changes in fruit antioxidant activity among blueberry cultivars during cold-temperature storage. , 2002, Journal of agricultural and food chemistry.

[6]  V. Tešević,et al.  Antioxidant properties of raspberry seed extracts on micronucleus distribution in peripheral blood lymphocytes. , 2009, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[7]  I. Rowland,et al.  Tracking (Poly)phenol components from raspberries in ileal fluid. , 2014, Journal of agricultural and food chemistry.

[8]  C. Bassi,et al.  Effects of Dietary Components on Cancer of the Digestive System , 2015, Critical reviews in food science and nutrition.

[9]  G. Blunden,et al.  Triterpenoids, Including One with Smooth Muscle Relaxant Activity, from Rubus idaeus (Raspberry) Leaves , 2006 .

[10]  M. Hansen,et al.  Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. , 1989, Journal of immunological methods.

[11]  S. N. El,et al.  Mind the gap—deficits in our knowledge of aspects impacting the bioavailability of phytochemicals and their metabolites—a position paper focusing on carotenoids and polyphenols , 2015, Molecular nutrition & food research.

[12]  M. Nair,et al.  Ursolic acid analogues: non-phenolic functional food components in Jamaican raspberry fruits , 2009 .

[13]  I. Rowland,et al.  Colon-available raspberry polyphenols exhibit anti-cancer effects on in vitro models of colon cancer , 2007, Journal of carcinogenesis.

[14]  Hee-Juhn Park,et al.  A dimeric triterpenoid glycoside and flavonoid glycosides with free radical-scavenging activity isolated from Rubus rigidus var. camerunensis , 2011, Archives of pharmacal research.

[15]  J. Gaforio,et al.  The differential localization of a methyl group confers a different anti-breast cancer activity to two triterpenes present in olives. , 2015, Food & function.

[16]  R. Kasai,et al.  Oleanane and ursane glucosides from Rubus species , 1992 .

[17]  J. Kennedy,et al.  Seed-coat anatomy and proanthocyanidins contribute to the dormancy of Rubus seed , 2011 .

[18]  S. G. Monteiro,et al.  Antimutagenic action of the triterpene betulinic acid isolated from Scoparia dulcis (Scrophulariaceae). , 2015, Genetics and molecular research : GMR.

[19]  T. Nohara,et al.  Two new triterpenoids from the seeds of blackberry (Rubus fructicosus) , 2016, Natural product research.

[20]  F. Sun,et al.  Bioactivity-Guided Fractionation of Physical Fatigue-Attenuating Components from Rubus parvifolius L. , 2013, Molecules.

[21]  J. Wolfender,et al.  LC/ES-MS analysis of triterpene glycosides: rapid estimation of the saponin content of dried berries of Phytolacca dodecandra , 1999 .

[22]  G. W. Robertson,et al.  A comparison of the composition of epicuticular wax from red raspberry (Rubus idaeus L.) and hawthorn (Crataegus monogyna Jacq.) flowers. , 2000, Phytochemistry.

[23]  F. Stampar,et al.  Composition of sugars, organic acids, and total phenolics in 25 wild or cultivated berry species. , 2012, Journal of food science.

[24]  J. Morales,et al.  Potential use of nanocarriers with pentacyclic triterpenes in cancer treatments. , 2016, Nanomedicine.

[25]  D. Graf,et al.  Anthocyanin-Rich Juice Lowers Serum Cholesterol, Leptin, and Resistin and Improves Plasma Fatty Acid Composition in Fischer Rats , 2013, PloS one.

[26]  S. Huttunen,et al.  Triterpenoid content of berries and leaves of bilberry Vaccinium myrtillus from Finland and Poland. , 2012, Journal of agricultural and food chemistry.

[27]  L. Ming,et al.  Triterpenoids from the Roots and Stems of Rubus alceaefolius , 2016, Chemistry of Natural Compounds.

[28]  M. Bakovic,et al.  Biologically Active Triterpenoids and Their Cardioprotective and Anti-Inflammatory Effects , 2015 .

[29]  M. A. Alhnan,et al.  Anti-glioma activity and the mechanism of cellular uptake of asiatic acid-loaded solid lipid nanoparticles. , 2016, International journal of pharmaceutics.

[30]  E. Weiderpass,et al.  Consumption of berries, fruits and vegetables and mortality among 10,000 Norwegian men followed for four decades , 2015, European Journal of Nutrition.

[31]  A. Dale,et al.  Genetic diversity of red raspberry varieties throughout the world. , 1993 .

[32]  C. Rao,et al.  Triterpenoids for cancer prevention and treatment: current status and future prospects. , 2012, Current pharmaceutical biotechnology.

[33]  P. Iversen,et al.  Polyphenol-rich juices reduce blood pressure measures in a randomised controlled trial in high normal and hypertensive volunteers , 2015, British Journal of Nutrition.

[34]  C. Hackett,et al.  Genetic and environmental effects influencing fruit colour and QTL analysis in raspberry , 2010, Theoretical and Applied Genetics.

[35]  S. Kitanaka,et al.  Triterpenoids from Hippophae rhamnoides L. and their nitric oxide production-inhibitory and DPPH radical-scavenging activities. , 2007, Chemical & pharmaceutical bulletin.

[36]  B. Burton-Freeman,et al.  Berries: anti-inflammatory effects in humans. , 2014, Journal of agricultural and food chemistry.

[37]  M. Sporn,et al.  Triterpenoids and rexinoids as multifunctional agents for the prevention and treatment of cancer , 2007, Nature Reviews Cancer.

[38]  J. W. Allwood,et al.  Novel colon‐available triterpenoids identified in raspberry fruits exhibit antigenotoxic activities in vitro , 2017, Molecular nutrition & food research.

[39]  A. Rodriguez-Mateos,et al.  Berry (poly)phenols and cardiovascular health. , 2014, Journal of agricultural and food chemistry.

[40]  S. Rohn,et al.  Phenolic profile and antioxidant activity of highbush blueberry (Vaccinium corymbosum L.) during fruit maturation and ripening , 2008 .

[41]  Reijo Karjalainen,et al.  Environmental and genetic variation of phenolic compounds in red raspberry , 2005 .

[42]  I. Rowland,et al.  In vitro and in vivo models of colorectal cancer: antigenotoxic activity of berries. , 2014, Journal of agricultural and food chemistry.

[43]  Ji-Hyung Seo,et al.  Anti-inflammatory effect of a standardized triterpenoid-rich fraction isolated from Rubus coreanus on dextran sodium sulfate-induced acute colitis in mice and LPS-induced macrophages. , 2014, Journal of ethnopharmacology.

[44]  D. Stewart,et al.  Assessing potential bioavailability of raspberry anthocyanins using an in vitro digestion system. , 2005, Journal of agricultural and food chemistry.

[45]  C. Bertsch,et al.  Fruit cuticular waxes as a source of biologically active triterpenoids , 2012, Phytochemistry Reviews.

[46]  C. Neto Ursolic Acid and Other Pentacyclic Triterpenoids: Anticancer Activities and Occurrence in Berries , 2011 .

[47]  C. Hackett,et al.  Over-seasons analysis of quantitative trait loci affecting phenolic content and antioxidant capacity in raspberry. , 2012, Journal of agricultural and food chemistry.

[48]  A. Collins,et al.  Protective effects of ursolic acid and luteolin against oxidative DNA damage include enhancement of DNA repair in Caco-2 cells. , 2010, Mutation research.

[49]  Rong-hong Zhang,et al.  Prodrugs of triterpenoids and their derivatives. , 2017, European journal of medicinal chemistry.

[50]  Takuya Suzuki,et al.  Regulation of the intestinal tight junction by natural polyphenols: A mechanistic perspective , 2017, Critical reviews in food science and nutrition.

[51]  I. Rowland,et al.  Persistence of Anticancer Activity in Berry Extracts after Simulated Gastrointestinal Digestion and Colonic Fermentation , 2012, PloS one.

[52]  G. Santoyo,et al.  A glimpse of the endophytic bacterial diversity in roots of blackberry plants (Rubus fruticosus). , 2016, Genetics and molecular research : GMR.

[53]  Wei Li,et al.  Triterpenoid saponins from Rubus ellipticus var. obcordatus. , 2009, Journal of natural products.