"Recent studies in vivo and in vitro indicate that aescin (β-escin) has significant antitumor activities. 
β-escin from A. hippocastanum inhibited chemically induced colon carcino-
genesis in rats, and in vitro exhibited cytotoxicity at 30 
µmol/L or above concentrations in colon cancer cell lines. β-Escin at 5 µmol/L also inhibited HT-29 colon cancer cell 
proliferation. β-escin induced cell cycle arrest at G1-S phase 
in part mediated by induction of p21WAF1/CIP1 and/or associated with reduced levels of Cdk2 and cyclins A and E 
complex; additionally, there was a lower phosphorylation of 
Rb [93]." (Shiyou Li et al.,2010)

"β-escin isolated from the seeds of A. chinensis var. 
chinensis induced apoptosis and caused a significant inhibition of HL-60 human acute myeloid leukemia cell and K562-
huamn chronic myeloid leukemia cell proliferation in dose- and time-dependent fasion. Morphological evidence of apoptosis, a significant increase of annexin V+ and PI- cells 
(early apoptotic) and apoptotic DNA fragmentation, were 
observed in K562 cells treated with β-escin. Flow cytometry 
analysis indicated that β-escin induced G1-S arrest and led to 
a significant accumulation of the sub-G1 population in HL-
60 and K562 cells 94, 95." (Shiyou Li et al.,2010)

"Aescin isolated from the seeds of A. chinensis var. wilsonii at a dose of 2.8 mg/kg had a rather high inhibition ratio 
(43.5 %) on mice H22 tumor growth in vivo. Aescin could 
induce significant concentration- and time-dependent inhibition of HepG (2) cell viability and 
induce cell cycle checkpoint arrest and caspase-independent cell death in HepG (2) 
cells 96." (Shiyou Li et al.,2010)

"An early investigation showed that two prosapogenins of 
21-O-tigloyl-22-O-angeloyl-R1-barrigenol (10) and 21-O-
angeloylbarringtogenol-C (18) from the acid hydrolysates of 
the HCSE of A. hippocastanum exhibited significant in vitro 
cytotoxicity with ED50 of 3.6 µg/mL and 3.0 µg/mL, respectively, in the human nasopharyngeal carcinoma 9-KB cell 
culture assay 27." (Shiyou Li et al.,2010)

"More recently, 28 individual saponins (72-99) from the 
fruits of A. pavia and six prosapogenins produced from these 
saponins were assayed in vitro for their cytotoxicity and inhibition of DNA topoisomerase I (TOP1) 61, 97 

Saponins 
94, 96 and 98 with two acyl groups at C-21 and C-22 
showed activity with GI50 of 0.175–8.71 µM against most of 
59 cell lines tested, which were from nine different human 
cancers including leukemia, non-small cell lung, colon, central nervous system (CNS), melanoma, ovarian, renal, prostate, and breast tumor cell lines. 

Aesculiosides IIc (79) and
IId (80) with only one acyl group at C-21 showed less activity while aesculiosides Ia-e (72-76) without acyl group 
showed no or weak activity 61. 

Aesculiosides IIa-k (77-
87), IIIa-f (88-93), IVa (95), IVb (97), IVc (99), saponins 94, 
96, 98 and six prosapogenins were also tested for their inhibition of DNA topoisomerase I (TOP1) and their activities 
against A549, PC-3, HL-60, PANC-1, and MRC cell lines. 

Most of the tested saponins and prosapogenins with acyl 
groups showed cytotoxic activity with different GI50 value. 

Sixteen cytotoxic aesculiosides 77-80 and 88-99 inhibited 
TOP1 catalytic activity by interacting directly with the free 
enzyme and preventing the formation of the DNA-TOP1 
complex. Interestingly, six prosapogenins, including 21-O-
angeloylproaescigin (13), 21,22-O-diangeloylprotoaescigenin 
(16), 21,22-O-diangeloylbarringtogenol-C (20), 21-O-angelyol-22-O-2-methylbutanoyl-R1-barrigenol, 21-O-angelyol-
22-O-2-methylbutanoylprotoaescigenin, and 21-O-angelyol-22-O-2-methylbutanoylbarringtogenol-C prepared from the 
acid hydrolysates of saponins 78, 96, 98, 95, 97, and 99, 
respectively, showed no TOP1 inhibitory activity, but had 
stronger cytotoxicity when compared to the related saponins 
97." (Shiyou Li et al.,2010)

"As far as cytotoxicity data are concerned the 
following observations can be made. Most studies 
were performed on a relatively narrow range of cell 
lines, usually from one to five, sometimes up to ten. 
The most notable exceptions in this respect are the 
reports, in which isolated compounds were assayed by 
National Cancer Institute (NCI) in anticancer drug 
discovery screen. For example, Zhang (Zhang and Li 
2007) tested triterpenoid saponins isolated from 
Aesculus pavia against a panel of 59 cell lines from 
nine different human cancers such as leukemia, non-
small cell lung, colon, CNS, melanoma, ovarian, renal, 
prostate and breast." (Podolak et al,2010)

"Zhang and Li screened five pre-selected saponins 
isolated from Aesculus pavia against a panel of 59 
cell lines from nine human cancers: leukemia, non-
small cell lung, colon, CNS, melanoma, ovarian, 
renal, prostate and breast. These saponins were 
polyhydroxyoleanenes with or without acyl groups 
at C-21 and/or C-22 and a sugar moiety composed of 
a-L-Araf, b-D-GlcAp, b-D-Galp. From the results 
obtained with a large array of cancer cell lines it seems that the presence of two acyl groups is 
important for cytotoxicity as non-acylated saponins 
showed no or only weak activity (Zhang and Li 
2007)." (Podolak et al,2010)

"The fruit of Aesculus hippocastanurn L. (horse chestnuts, Hippocastanaceae) has been 
used as an herbal remedy for the treatment of mammary indurations and cancer (1). 
Prior phytochemical studies (2-5) on this plant have yielded a saponin, aescin, which is 
a mixture of the acylated glycosides of protoaesigenin and barringtogenol-C, with 
angelic, tiglic, and acetic acids as the acyl groups. As a result ofour continuing searches 
among medicinal plants for novel, naturally occurring, potential antitumor agents, the 
acid hydrolyzed product of an n-BuOH extract of the fruits of A. hippocastanurn was 
found to show significant in vitro cytotoxicity in the 9-KB (human nasopharyngeal car- 
cinoma) cell culture assay (6). Bioassay-directed fractionation of the foregoing active ex- 
tract led to the isolation and characterization of two cytotoxic sapogenols, the new hippocaesculin
 [1, ED50 (KB)=3.6 pg/ml] and the known barringtogenol-C 2 1-angelate 
12, ED,, (KB)=3.0 pg-mll. Compound 2 was previously isolated from the leaves of 
Pittosporum tobira (7); however, its cytotoxicity is revealed for the first time." (konoshima1986)
      

Phytochemicals

Bark:

"The bark of Aesculus hippocastanum, in common with the seeds, has been described as containing the saponin mixture ‘escin’ ³. Coumarin glycosides, including esculin, scopolin and fraxin, and their respective aglycones, esculetin, scopoletin and fraxetin, are also present in the bark of A. hippocastanum (Figure 4), in contrast to seed tissues, in which these compounds have not been detected. The flavonoid glycoside quercitrin (Figure 2), and its corresponding aglycone have also been detected in bark tissues. Additional compounds, including allantoin, sterols, leucocyanidin, leucodelphinidin, catechol tannins and alkanes have also been described as occurring in bark tissues. ³" ^{(wilkinson1999)}

Leaves:

"In common with the bark of A. hippocastanum, leaf tissues contain the coumarin glycosides scopolin, fraxin and esculin (Figure 4) [3]. A range of flavonoid glycosides of quercetin (e.g. quercitrin, rutin, isoquercitrin and quercetin 3-arabinoside) and the corresponding glycosides of kaemperfol have also been detected in leaf tissues (Figure 2). In addition to these glycosides, escin has been detected (but only in trace amounts), as well as leucanthocyans, cis,trans-polyprenols, amino acids, fatty acids and sterols (sitosterol, stigmasterol and campesterol)." ^{(wilkinson1999)}

• Triterpene saponins ^[PDR]
• Hydroxycoumarins: "chief component is aesculin, in addition fraxin and scopolin" ^[PDR]
• Flavonoids: "including rutin, quercitrin, and isoquercitrin" ^[PDR]
• Tannins ^[PDR]

"Twenty-one carotenoids have been identified from Aes- culus. An early investigation on the leaves and pollen of the three species A. turbinata, A. pavia, and A. parviflora by HPLC analysis indicated the presence of seven carotenoids β-carotene (162), zeaxanthin (163), α-carotene (167), lutein (168), capsanthin (176), capsanthin 5,6-epoxide (177), and capsorubin (178) [74]. A high concentration of α-carotene was found in the leaves and keto hydroxyl carotenoids with a pentanuclear ring structure (capsanthin, capsanthin 5,6- epoxide, and capsorubin) were identified in the pollen. These keto hydroxyl carotenoids were proposed to have special chemotaxonomic significance for the genus Aesculus. Later, in 2000, 16 carotene derivatives were detected in buds, pol- len, and petals of A. hippocastanum and A. pavia by HPLC analysis with a diode array detector utilizing authentic sam- ples as the references. These carotenoids are β-carotene (162), β-cryptoxanthin (164), violaxanthin (165), 9-cis-violaxanthin (166), lutein (168), 13-cis-13'-cis-lutein (169), lutein 5, 6-epoxide (170), neolutein C (171), β-citraurin (172), aesculaxanthin (173), 9-cis-aesculaxanthin (174), 13-cis-aes- culaxanthin (175), 9'-cis-neoxanthin (179), 9-cis-9'-cis-neox- anthin (180), neochrome (181), and luteoxanthin (182) [75]." ^{(Shiyou Li et al.,2010)}

Seed Phytochemicals

"Horse-chestnut seeds, as all others seeds, are natural products whose chemical composition is a very complex matrix. They contain a lot of different molecules and analytes, the majority of which are polysaccharides (both starches and non-starches), proteins, lipids, mineral salts and many minor components among others, with homogeneous or localised distributions in different districts, all strongly interacting with each other in a synergistic way to form very complex structures." ^{(baraldi2007)}

"A number of other products have been isolated from chestnut seeds, i.e., coumarin derivatives (aesculin, fraxin, scopolin), essential oils (oleic acid, linoleic acid), and tanins (leucocyanidine, proanthocyanidin A2) (4)." ^{(kapusta2007)}

"It is interesting that different organs of this species contain distinctive classes of the main bioactive principle: aescin in the seeds, essential oils in the leaves, and flowers and coumarins in the bark." ^{(kapusta2007)}

"For completeness, our experimental data about the com- position of common Mediterranean horse-chestnuts, can be compared with some literature values reported by Par- mar and Kaushal (1982), even though they worked with Aesculus indica seeds from fresh fruits. Aesculus indica is a very common botanical species and largely diffused in Himalayan forests, also pertaining to Hippocastanaceae family, with some distinctive features and characters from each others.
Probably, this comparison can result in a limited useful- ness because of different uses of AHP or AHH and Aescu- lus indica seeds, being the latter foodstuff still largely consumed by local populations of Himachal Pradesh region. Starch of seeds (about 40% on fresh fruits, with a ratio amylopectin:amylose =/~3:1) was recommended as famine food for extending bread flour (or other uses), after removal of bitter characters." ^{(baraldi2007)}

• "...powdered hydroalcoholic extracts of the seeds contain 16 to 20 percent triterpene glycosides (a class of saponins), calculated as aescin (escin). Aescin... is believed to be the main active constituent of horse chestnut seed extract (Schulz, Hänsel, and Tyler, 2001)." ^{[Barrett HCTHR]}
• "Triterpene saponins (3-5%): The triterpene saponine mixture known as aescin (also escin) consists of diacylated tetra-and pentahydroxy-beta-amyrin compounds...." ^[PDR]
• Flavonoids: "in particular biosides and triosides of the quercetins" ^[PDR]
  
• Oligosaccharides: "including 1-kestose, 2-kestose, stachyose" ^[PDR]
  
• Polysaccharides: "starch (50%)
• Oligomeric proanthocyanidins, condensed tannins: (only in the seed-coat)" ^[PDR]
  
• Fatty oil (2-3%) ^[PDR]

"Seeds of A. hippocastanum have also been described as containing starch (40–50% weight) [6], sugars, proteins (specifically the globulin, hippocastanine, containing L-(1)lysine and L-(1)-tryptophan), a fatty oil (containing oleic, linoleic, linolenic, stearic and palmitic acids) and purines (adenosine, adenine and guanine). [5]" ^{(wilkinson1999)}

"The saponins present in these seeds render them very bitter, disagreable and unedible for humans. On the con- trary, they can be consumed for human uses after removing the bitterness of grounded flour, by soaking it in water for about 12 h. The bitter components get dissolved in water and removed when the water is decanted. The remaining slurry can be destined for cuisine manipulations, and is generally taken as a non-cereal-starch food. As reported by Parmar and Kaushal (1982), the Aesculus indica seeds, which constitute the edible portion of the fresh fruits, con- tain about 50.5% moisture. The total sugars content is 5.58% (11.0% on dry matter basis), combining the reducing (4.59%) and non-reducing sugars (0.94%), respectively. The protein and mineral contents are 0.388% and 1.934%, respectively, on fresh samples, that become 0.768% (pro- teins) and 3.83% (minerals) on dry samples. Therefore, we can assert that Aesculus indica seeds are less proteinic and less glucidic with respect to AHP and AHH seeds sam- ples here tested." ^{(baraldi2007)}

"Coumarins isolated from Aesculus usually have simple structures." ^{(Shiyou Li et al.,2010)}

"The seeds of Aesculus also contain a number of long fatty chain compounds." ^{(Shiyou Li et al.,2010)}

Aescin (Seeds)

"The principal extract and medicinal constituent of Aesculus hippocastanum (horse chestnut) seed is aescin, a mixture of triterpenoid saponin glycosides. It can be fractionated into beta-aescin, an easily crystallizable mixture, and alpha-aescin, which is water-soluble." ^[HPEP]

"The total saponin content of seeds, often expressed as ‘(a)escin’ (Figure 1), actually consists of α-escin and β-escin, the latter of which is, in turn, composed of more than 30 derivatives of the triterpenoids, protoaescigenin and barringtogenol C. These compounds are primarily found in the seed cotyledons (they can constitute up to 28% of the weight of the dry seeds), but have also been detected in the seed integument, the bark, buds, leaves and the immature fruit pericarp of A. hippocastanum [5]." ^{(wilkinson1999)}

"Aescin is fairly soluble in water but is poorly soluble in lipid solvents." ^{[Schulz RP]}

"CAN cautions that aescin is nephrotoxic. Side effects include GI disturbance, impaired liver function, mild nausea, shock, spasm, urticaria, and vomiting. Should be avoided by patients on blood-thinning therapy, with hepatic or renal impairment, or lactating or pregnant. Large doses of saponins can be fatally hemolytic in animals. LD50s range for aescin from 134 to 720 orally in mice, rats, and guinea pigs. On ipr administration, the total saponin fraction (LD50 = 46.5 mg/kg ipr mouse) was less toxic compared to isolated aescin (LD50 = 9.5 mg/kg ipr mouse) (CAN). LD50 of seed extract 990 mg/kg orl mouse, 2150 orl rat, 1530 orl rbt, 130 orl dog."^{[HMH Duke]}

Flavonoids (Seeds)

"Flavonoids and their derivatives are also one of the main components of the genus Aesculus. A total of 49 flavonoids including flavonols and their glycosides (100-126), flavanones, and flavanone derivatives (127-148) were iso- lated and identified from the seeds of Aesculus." ^{(Shiyou Li et al.,2010)}

"In the present work, the main flavonoids from horse chestnut seeds were isolated and their structures established with spectral methods. Seven glycosides were isolated, out of which six (2, 3, 4, 7, 11, 13) were previously reported and one (9) was identified as a new tamarixetin 3-O-[β-D-glucopyranosyl(1→3)]-O-α -D-xylopyranosyl-(1→2)-O-β-D-glucopyranoside." ^{(kapusta2007)}

"Thirteen compounds could be identified in the profile, out of which di- and triglycoisdes of quercetin and kaempferol were the dominant forms and their acylated forms occurred in just trace amounts. The total concentration of flavonoids in the powdered horse chestnut seed was 0.88% of dry matter. The alcohol extract contained 3.46%, and after purification on C18 solid phase, this concentration increased to 9.40% of dry matter. The flavonoid profile and their content were also measured in the horse chestnut wastewater obtained as byproduct in industrial processing of horse chestnut seeds. The total flavonoid concentration in the powder obtained after evaporation of water was 2.58%, while after purification on solid phase, this increased to 11.23% dry matter. It was concluded that flavonoids are present in a horse chestnut extract in a relatively high amount and have the potential to contribute to the overall activity of these extracts. Industrial horse chestnut wastewater can be used to obtain quercetine and kaempferol glycosides for cosmetic, nutraceutical, and food supplement industries." ^{(kapusta2007)}

"In the present research, we also studied the flavonoid profile and concentration in the powder obtained after evaporation of water from horse chestnut wastewater (WWHC). The large volume of WWHC remains in the process of precipitation of aescin from the horse chestnut extracts. This is waste byproduct still containing some amount of saponins and flavonoids, which can be commercially useful." ^{(kapusta2007)}

"The flavonoid profile of WWHC was identical to the profile of HCE, with the total concentration of 2.54% dry matter. One-step purification of this fraction on the solid phase produced WWHCF, the product with flavonoid concentration of 11.23% dry matter. This has been a quite high amount, and the product can have commercial value. It can be used in the pharmaceutic, cosmetic, and food industries. Eleven percent concentration of flavonoids makes the product an attractive source for the nutraceutic industry as a high-flavonoid supplement. As shown above, horse chestnut flavonoids are the mixture of quercetin and kaempferol glycosides, the most desired flavonoids in our diet due to their antioxidant activity. Quercetin and kaempferol were documented to be the most potent antioxidant of all flavonoids, (10) and their daily consumption is estimated to be about 25 mg per day (11). Further increase of this amount in the diet can be obtained by flavonoid supplementation, and WWHCF can be a good source of this. The economic evaluation of this source should be performed. It seems logical that the WWHC, instead of being dried and powdered, can be simply passed through a solid-phase column (reversed-phase C18 or Amberlite XAD4), which retains flavonoids and lets saccharides and other strongly polar components of the WWHC matrix go through." ^{(kapusta2007)}

"It can be concluded that the flavonoid content in horse chestnut seed seems to be high enough to contribute to overall activity of the extracts. The high content of flavonoids in industrial horse chestnut wastewater and the ease of their condensation and purification makes this byproduct a promising source of quercetin and kaempferol glycosides to be used in the cosmetic and food additive-producing industries." ^{(kapusta2007)}

"Nine flavonol oligosides of quercetin and kaempferol with glucose, xylose, and rhamnose as sugars were isolated from the seeds of Aesculus hippocastanum L. (Hippocastanaceae). Five of them are new compounds (2 trisaccharides, 1 bisdesmoside, a nicotinic and a indolinone hydroxyacetic acid ester of the bisdesmoside)." ^{(Hubner,1999)}

"Extracts of the seeds of Aesculus hippocastanum L. (horse chestnut; Hippocastanaceae) are widely used to successfully treat chronic venous insufficiency (1), (2). The main active constituent is considered to be aescin, a mixture of several saponin esters of the oleanane type (1), (3). However, some authors believe that the flavonoids contribute to the observed activity of horse chestnut seed extracts (3), (4)." ^{(Hubner,1999)}

"The partial lack of knowledge about the polyphenolic fraction of this widely used crude drug has led to the following re-examination of the flavonoids of the seeds of A. hippocastanum." ^{(Hubner,1999)}

"A quantitative determination of the flavonoids according to Betulae folium of the European Pharmacopoeia (1997) gave a total flavonoid content of 0.29% (calculated as rutin)." ^{(Hubner,1999)}

"In conclusion, the seeds of A. hippocastanum contain a series of quercetin and kaempferol di- and triglycosides, some of which have unusual acyl substituents. The overall content of ca. 0.3% (calculated as rutin) is low. Hence, a contribution of the flavonol glycosides to the therapeutic efficacy of the crude drug seems unlikely." ^{(Hubner,1999)}

Sugars (Seeds)

"The values obtained for total glucides content in these natural products was in the range 14.3% (AHH)–15.2% (AHP). As previously mentioned in the experimental sec- tion, our analysis methodology permit us to determine the two most important monosaccharides (and appar- ently, the only species in these matrices), such as glucose and fructose. However, we observe that while glucose concentration is almost the same on the two sample groups (6.8% for AHP and 6.9% for AHH), fructose con- tent is well differentiated (8.4% for AHP and 7.4% for AHH).
Actually, we are unable do determine the presence of other sugars, such as the most common sucrose (the disac- charide based on glucose and fructose condensation). Nev- ertheless, we are obliged to mention that, generally, edible chestnut seeds mainly contain sucrose (about 10–30%), and only few traces of free monosaccharides (about 0.1–0.5%) (De La Montana Miguelez et al., 2004)." ^{(baraldi2007)}

Fatty Acids (Seeds)

"Therefore, the aims of this work were to study some characters of chemical composition and the relevant classi- fication of seeds from two most wide spread varieties of Aesculus hippocastanum L., such as pure species, with white flowers (AHP), and a common hybrid species with pink flowers (AHH). Both these common Hippocastanum trees are growing in the mediterranean regions, and their luxuri- ous summer flowering is very showy." ^{(baraldi2007)}

"Lipids represent a significant fraction on these original products (4–5% of the dry mass)" ^{(baraldi2007)}

^{(baraldi2007)}

Epicuticular Leaf Waxes

"Ae. hippocastanum leaves (102 g) contained an extractable epicuticular wax (241 mg) of about 0.63% dry wt., which calculated for 22 µg wax per cm² leaf surface area or 4540 µg wax per one leaf. This wax of mature leaves consisted, like the wax of C. sativa (Castanea sativa), of homologous series of wax lipids and in addition greater amounts of triterpenoids (54% of the wax). Main lipid classes are aldehydes (13%), fatty acids (8 %), primary alcohols (7%), wax esters (7%), hydrocarbons (5%) and addi­tionally acetates (5%). Composition and yield of the individual lipid classes are listed in Table I. The compositions of the homologous series are sum­marized in Table II.
All these lipid classes have the same chain lengths as found for C. sativa and show no main component dominating. The distribution patterns of these lipids are therefore very even.
Triterpenols were found to be the dominating wax compounds. ß-Amyrin, a-amyrin and lupeol were again predominant in a relative ratio of 3:2.2:2.2. These triterpenols were present also esterified with fatty acids (2% of the wax). Fur­thermore, friedelanol and friedelanone were iden­tified by GC-MS and comparison with authentic samples [6 , 7]. The latters were found in concentra­ tions of about 1.5% of the wax, each...." ^{(P.-G. Gülz et al.)}

"The SEM figures of mature leaves of Ae. hippo­castanum show no trichomes on the abaxial as well as on the adaxial epidermis. Both leaf surfaces are characterized by numerous cuticular lamellae (Fig. 2, A and B). The cuticular lamellae are found to be linear (Fig. 2,C) and also wavelike (Fig. 2, D). All epidermal cells are covered with a continuous wax layer without any wax sculptures or crystalloids. After washing the leaves with chlo­roform all wax was extracted. The remaining cuti­ cular lamellae are clearly visible on the abaxial and adaxial epidermal cells...." ^{(P.-G. Gülz et al.)}

"C. sativa and Ae. hippocastanum are systemati­ cally not very closely related plant genera. How­ever, they have a rather similar wax composition. They contain the same homologous series of wax lipids, but with individual distribution patterns. No one of these lipid classes has one main compo­nent dominating. In addition to the wax lipids, both plant waxes contain mixtures of triterpenols. In C. sativa wax ß-amyrin, a-amyrin and lupeol are found. In Ae. hippocastanum wax the same tri­terpenols are analyzed and additionally friedelanol and friedelanone. Both leaf waxes contain wax lipids and triterpenols with no main component in a dominating concentration. Therefore, wax crys­talloids on the leaf surfaces of these two plants are not expected." ^{(P.-G. Gülz et al.)}

Folkore

"The common name "horse chestnut″ may have come from the uses of seeds for horses to treat overexertion or coughs by Turks and Greeks [18]. Dated from the early 18th century, the horse chestnut has a therapeutic property for anti-fever [5]. In Europe, the bark and leaves of A. hippocastanum have been employed as an astringent to treat diarrhea and hemor- rhoids [6]." ^{(Shiyou Li et al.,2010)}

• The drug is generally provided by the ripe seed, as described in German and Spanish pharmacopeias, while Portuguese pharmacopeia also includes the bark. ^{[Bajaj MAPS 7]}
• Some kind of horse chestnut decoction was drunk in Essex for lumbago, spoken of there as rheumatism (Newman & Wilson)." ^{[DPL Watts]}
• "Elsewhere, including Spain (H W Howes), it is piles that is reckoned to be cured or prevented by carrying them around (W B Johnson; Tongue; Fogel), and that is interesting, because it is known that extracts of horse chestnut are rich in Vitamin K, and so is useful in treating circulatory disorders like piles, varicose veins and chilblains (Conway)." ^{[DPL Watts]}
• "A fluid extract made from the nuts is also used to protect the skin from the harmful effects of the sun (Schauenberg & Paris)." ^{[DPL Watts]}
• "Another usage of conkers was as a snuff to cure catarrh and headache. The Pennsylvania Germans used it that way (Fogel), but this was quite an early habit (see Thornton), and the idea was to grate them up and use the powder to make one sneeze. Apparently it was recommended not only as a powder but also as an infusion or decoction to take up the nostrils. The leaves and flowers have occasionally been used, too (and so has the bark)." ^{[DPL Watts]}
• "The leaves are narcotic; an infusion of them has been used for insomnia(Conway)" ^{[DPL Watts]}
• "A tincture of the flowers is sometimes given for rheumatism (Perry. 1972)." ^{[DPL Watts]}
• "The bark has been used for fevers, and externally, for ulcers (Wickham)" ^{[DPL Watts]}
• Several French works published between 1896 and 1909 reported successful outcomes in the treatment of hemorrhoidal ailments. ^{[Schulz RP]}

"The bark is no longer in common use, but has a folk medicinal application as a febrifuge, and as an astringent in cases of diarrhoea and haemorrhoids [4]. Decoctions of the bark are also used, albeit rarely, for the topical treatment of skin disorders, such as sores, lupus and ulcers [4]. The bark has also previously been used as an anti-malarial agent, as a cinchona substitute, but this practice is no longer continued [3]. Horse chestnut leaf preparations are used in folk medicine to treat coughs, rheumatism and arthritis, although the underlying phytochemical basis for these applications has not been determined [3]." ^{(wilkinson1999)}

Cultivation

"Prefers a deep loamy well-drained soil but is not too fussy tolerating poorer drier soils[11, 200]. Tolerates exposed positions and atmospheric pollution[200]. A very ornamental and fast-growing tree[1, 4], it succeeds in most areas of Britain but grows best in eastern and south-eastern England[200]. Trees are very hardy when dormant, but the young growth in spring can be damaged by late frosts. The flowers have a delicate honey-like perfume[245]. Trees are tolerant of drastic cutting back and can be severely lopped[200]. They are prone to suddenly losing old heavy branches[98]. The tree comes into bearing within 20 years from seed[98]. Most members of this genus transplant easily, even when fairly large[11]. Special Features: Attractive foliage, Not North American native, Naturalizing, Blooms are very showy."^[PFAF]

"Presowing treatment of seeds with cobalt nitrate increased drought resistance of horse chestnut (Aesculus hippocastanum L.) from the Donets Basin in southeastern Europe (87)." ^{[Barker HPN]}

"In conclusion, our results suggests that in horse-chestnut, where natural variation is large and vegetative propagation through natural cross-hybridisation and grafting is very easy, the traditional method of selection variety may give good results in terms of selected fruits and seeds productions, also in relation to the optimisation procedures for technological transferability, and to obtain the best performances for bio-available supplements and specific salutary targets." ^{(baraldi2007)}

A single exotic pathogen that has appeared in Europe in the last decade, has devastated the European horse chestnut (Aesculus hippocastanum). "Trees were observed to be suffering from a new form of bleeding canker on their stems which ultimately kills them, firstly in continental Europe and more recently across the UK as well. The causal agent of the disease was identified as a new species of pathogenic bacterium; Pseudomonas syringae pv. aesculi. It is thought that this bacterium originated in India on the Indian horse chestnut, and was probably introduced to Europe via the plant trade (as highlighted by Brasier 2008).... results have shown that there is only one strain of this organism across the whole of Europe, which suggests that the outbreak is probably the result of a single introduction event." ^{[Fenning COWF]}

"...A. hippocastanum could be suggested as an appropriate biomonitor for Pb atmospheric pollution, and for Cu in highly polluted areas." [Anicic, 2011]

Propagation

"Seed - best sown outdoors or in a cold frame as soon as it is ripe[11, 80]. The seed germinates almost immediately and must be given protection from severe weather[130]. The seed has a very limited viability and must not be allowed to dry out. Stored seed should be soaked for 24 hours prior to sowing and even after this may still not be viable[80, 113]. It is best to sow the seed with its 'scar' downwards[130]. If sowing the seed in a cold frame, pot up the seedlings in early spring and plant them out into their permanent positions in the summer."^[PFAF]

"Propagation is generally by seed, which fall in autumn, remain dormant over winter, and germinate in spring, at least in Liguria (Italy). In the first year plants have either two or four leaves. The vegetative growth of such plants returns the following year. The plant may also be propagated by cutting; in this case the specimens have the same morphological and physiological characteristics as the parent plant." ^{[Bajaj MAPS 7]}

Bioaccumulation

"The mean concentrations of heavy metals (Pb, Cd, Zn and Cu) in the unwashed and washed leaves, bark and soils of A. hippocastanum from different sites are presented.... A comparison of concentrations of heavy metals in the leaves, barks and soils reveals that the values in the urban roadside are higher than other sites due to higher human activity and higher vehicular traffic. A similar finding is reported by Tam et al. (1987) from Hongkong who found a significant correlation between traffic density and Pb, Cu and Zn concentrations. The heavy metal concentration in the bark and soils are higher than in the leaves as in our findings." ^{(Yilmaz,2006)}

"The highest lead concentration was found in the urban roadside soils (6.745 µg g-1) as compared to the suburban soils (0.812 µg g-1)....In the A. hippocastanum leaves the highest values of lead in the urban roadside samples (0.119 µg g-1),....Lead concentration of the bark too as expected was high in the urbanroadside samples (0.628 µg g-1)," ^{(Yilmaz,2006)}

"The Cd values in our samples in general are very low, but highest values were recorded in the urban roadside unwashed samples (0.068 µg g-1). The values for unpolluted natural environments should lie between 0.01-0.03 µg g-1 (Allen, 1989)....The reason for the low values could be due to a low atmospheric deposition in this region because of long lasting rainy season." ^{(Yilmaz,2006)}

"In A. hippocastanum the Zn concentrations measured during this study were between 0.374 - 0.532 in the washed leaves, 0.391-0.591 in the unwashed leaves, 0.406- 0.660 in the bark and 2.196 and 4.598 µg g-1 in soil....By following the Zn pollution criteria levels of Dmuchowski and Bytnerowicz (1995), we can conclude that Thrace region does not show Zn pollution....In the uncontaminated soils, the total Zn content is said to lie between 10-300 µg g-1 (Freedman & Hutchinson, 1981). In these soils, total Zn is constant in the soil profile, but extractable Zn generally decreases with depth because of decrease in organic matter. The higher amount of the Zn in our soils can be attributed to the high organic matter content." ^{(Yilmaz,2006)}

"The soil Cu can vary greatly among the sites at the same location and in different locations. The Cu values of soil samples supporting A. hippocastanum lie between 0.5171 and 1.1165 µg g-1,in the unwashed leaves Cu content ranges between 0.322-0.466, in the washed leaves between 0.256 and 0.387 and in the bark between 0.3451-0.117 µg g-1. In normal uncontaminated soils Cu content varies between 2-100 µg g-1 (Freedman & Hutchinson, 1981). Our values are low."

"Washing the leaves significantly reduced the lead, cadmium, zinc and copper concentrations. The reduction for the lead was 57.14 percent for the urban roadside, 27.66 for the city centre and 13.04 for the suburban area, whereas for cadmium these values were 97.06, 50.0 and 0.0 % respectively. In the case of zinc the reduction was 10.43 % in the urban roadside, 3.36 % in the city centre and 4.35 % in the suburban areas, and for copper it was 16.95 % in the urban roadside, 17.39 % in the city centre and 20.50 % in the suburban areas. The percentage removal was the highest in the urban roadside (57.14 %) as compared to suburban. The reason for this is differences in the atmospheric deposition of these metals and high and low traffic densities. This data coincides with that of Al-Shayeb et al. (1995) who reported a removal of 26-68 % of Pb by washing." ^{(Yilmaz,2006)}

"The atmospheric deposition of heavy metals in the present study area is not very high, the concentration of heavy metals in A. hippocastanum did not exceed the upper limit." ^{(Yilmaz,2006)}

"The regression analysis data shows (Fig. 6) that our values for soils and plants are significant for Pb, Zn and Cu, but not for Cd. We can thus conclude that with an increase in the amount of the heavy metals in the soil, their uptake by plants also increased." ^{(Yilmaz,2006)}

"A. hippocastanum is widely distributed in Europe and is used as a roadside ornamental tree. In accordance with the data presented here this tree possesses all the characteristics for its selection as a biomonitor." ^{(Yilmaz,2006)}

"This study gives an evidence for the seasonal accumulation of Cr, Fe, Ni, Zn, and Pb in leaves of A. hippocastanum and Tilia spp. (except Zn) sampled in the Belgrade urban area. During the studied time span, Pb concentrations in the leaves showed a decreasing trend (more regular for A. hippocastanum), being in accordance to the bulk atmospheric deposition data. Also, the temporal concen- tration trend for the Cu in A. hippocastanum was decreasing and in accordance with the Cu trends in the bulk atmospheric deposition at the site with the highest atmospheric Cu loading. No agreement was observed between the accumulation trend of Cr, Fe, Ni, and Zn in leaves and the bulk deposition rates, i.e. the elements content in the leaves did not reflect atmospheric deposition directly.
Accordingly, A. hippocastanum could be suggested as an appropriate biomonitor for Pb atmospheric pollution, and for Cu in highly polluted areas." ^{(Anicic,2011)}

"The concentrations of cadmium, copper, lead and zinc have been measured in the leaves of a deciduous tree the horse chestnut (Aesculus hippocastanum L.) over the period of their lifetime (7 months). The average concen- trations for the total sample based on ash weight are." (txg g l) cadmium, 0.197; copper, 129; lead, 294; and zinc, 299. The temporal trends in the concentrations of the metals can be related to their dominant source. Cop- per and zinc concentrations are highest in the new leaves and decrease with time, suggesting the main source of the elements are uptake from the soil. The decrease occurs partly because of dilution by leaf material as it increases over the growing period. In the case of zinc, however, aerial deposits appear to be also a significant source. Lead concentrations, on the other hand, show an increase with time, which can be related to increasing deposits from aerosol lead arising from the combustion of petrol lead. The increase is enough to offset the dilution effect. For cadmium there is no significant trend, but the ten- dency is a decrease with time. It is not possible, however, to distinguish between soil uptake and aerial deposit as both are small compared with increase in leaf material." ^(kim1994)

"The measure of the maturity of the leaves in terms of the amount of internal consolidation ( g c m 2) is lowest in samples collected when the leaves first opened, and increases steadily until January-March. Ash weight concentrations of copper and zinc show the opposite trends, and decrease (in a non-linear manner) with the age of the leaves. Highly significant negative correla- tions between the variable ( g c m -2 x 105) and the con- centrations of copper and zinc suggest that growth of the leaves causes dilution in the concentrations of cop- per and zinc, but also there may be some loss of the metal ions out of the leaves. Growth of the leaves may account for about 60--70% of the reduction in copper and zinc concentrations with time." ^(kim1994)

"The fact that copper and zinc concentrations are highest when the leaves first open implies that these metals are primarily derived from the soil and that most of the copper and zinc is translocated into the leaves prior to their unfolding. In the case of zinc, how- ever, as the leaves grow an atmospheric contribution becomes important, offsetting to some extent the dilu- tion due to leaf growth. The association of copper and zinc with developing leaves is probably a result of the roles that these two metals have in plant protein and carbohydrate metabolism." ^(kim1994)

"Ash weight lead concentrations are lowest when the leaves first appear, and increase in a linear (and highly significant) manner with the age of the leaves. The source of this lead is likely to be petrol combustion, and it is probable that direct deposition of the lead-rich aerosols on the leaves (rather than translocation of lead from the roots) is the predominant enrichment path- way. The rate of lead accumulation is sufficient to negate any dilution effect that might be caused by leaf growth." ^(kim1994)

"No clear (statistically justified) trend is evident which links cadmium concentrations in horse chestnut leaves with the age of the leaves. Additionally, cadmium con- centrations in the leaves are uncorrelated with those of copper, lead or zinc. However, a 'very significant' nega- tive correlation between cadmium concentrations and the variable (g cm 2 x 10 5) suggests that growth of the leaves may cause some dilution in cadmium concentra- tions. It is possible that, if cadmium is accumulating on/in the leaves, the rate of this accumulation is suffi- cient to partially compensate for the dilution effect caused by the leaves growing. Unlike the results for copper, lead and zinc, the results for cadmium do not clearly identify probable sources or enrichment path- ways of that metal to leaves." ^(kim1994)

"Leaves collected from the three sites situated near main roads (sites 1, 4 and 5) have statistically more cadmium, copper, and lead than leaves collected from the other sites, implying that traffic is a source of cad- mium, copper and lead to horse chestnut leaves. The atmospheric source of zinc may derive from weathering of galvanised iron roofs, which are relatively common in New Zealand." ^(kim1994)

AESCULUS BUCKEYE

"Large shrub or tree. Leaf: palmate, leaflets 5–7[9]. Flower: petals 4[5], >> sepals. Fruit: capsule leathery. Seed: 1, large.
± 15 species: northern hemisphere. (Latin name for a sp. of oak)" ^[Jepson]

Local Species;

1. Aesculus hippocastanum - Horse-chestnut [Cultivated]

"Aesculus L. is a genus of the family Hippocastanaceae containing 12 species of deciduous trees and shrubs in the northern hemisphere, primarily in eastern Asia and eastern North America, with one species native to Europe, and two to western North America [1-4]." ^{(Shiyou Li et al.,2010)}

"Aesculus hippocastanum has been cultivated since 1576 as an ornamental plant [17] while A. chinensis var. chinensis has been planted in temples and homes for several centuries. At present, all species are cultivated, with at least 27 commercial cultivars supplied by at least 95 nurseries in the United States [17]." ^{(Shiyou Li et al.,2010)}

"There are two Eurasian species commonly used in medicine: A. hippocastanum (common horse chestnut) and A. chinensis var. chinensis (Chinese horse chestnut)." ^{(Shiyou Li et al.,2010)}

"To date, over a hundred species or varieties in Aesculus have been described because of great morphological variations due to natural pollination among the species. In his monograph of Aesculus, Koch (1857) recognized 13 species in four subgenera, Hippocastanum, Pavia, Calothyrsus, and Macrothyrsus [12]. Based on bud viscidity, fruit exocarp ornamentation, flower color, and petal morphology, the 13 species were grouped into five sections [1, 2, 13]. All new species discovered in the last several decades are not recognized and thus most authors recognized 13 species in the genus until A. wilsonii Rehder was recently treated as a variety under the species A. chinensis Bunge [3, 14]. The 12 currently recognized species in the genus Asculus are grouped in the five sections...." ^{(Shiyou Li et al.,2010)}

Chemotaxonomy

"....Some chemotaxonomical analysis has been conducted [15]. Interestingly, A. pavia in the Section Pavia has tritepernoid saponins with an oligosaccharide chain at C-3 of the aglycone with an α-arabinofuranosyl unit affixed to C-3 of the glucuronic acid while Eurasian species in Section Aesculus and Section Calothyrsus have saponins with a trisaccharide chain at C-3 of the aglycone with a β-glucopyranosyl unit attached to C-4 of the glucuronic acid [16] (see Triterpenoid Saponins for details). This significant chemical difference suggests that triterpenoid saponins may provide important clues in understanding the systematics and evolution of the genus Aesculus." ^{(Shiyou Li et al.,2010)}

Use of Related Sp.

"Seeds of North American species (e.g., A. pavia, A. flava, A. glabra, and A. californica), known as buckeyes, were used by Native Americans to tranquilize fish to make them easier to catch [22]. Extracts from the seeds were used to treat earaches, sores, colic, sprains, and chest pains [23]. Powdered bark was sometimes used to alleviate toothaches and ulcer pain [24]." ^{(Shiyou Li et al.,2010)}

"The genus Aesculus has 12 species, but only A. hippocas- tanum and A. chinensis var. chinensis, two Eurasian species, are officially recognized sources of herbal products in tradi- tional medicine. HCSE or aescin from A. hippocastanum has shown satisfactory evidence for clinically significant activity in chronic venous insufficiency, hemorrhoids, and post- operative oedema, largely due to its anti-inflammatory prop- erties, which were well demonstrated by in vitro and in vivo assay. However, the mechanism of action of the Chinese horse chestnut is poorly understood and further investigation is necessary." ^{(Shiyou Li et al.,2010)}

Aesculus californica - California buckeye

"Nuts processed into mush and served with pit oven-cooked deer meat, seafood, roasted peppernuts, and contemporary picnic foods within the living memory of Kashaya Pomo individuals (Ortiz 1989a: 25)" ^(Ortiz,GTR)

"Aesculus californica (Spach.) Nutt. Berraco; chattYa (M), chatch (R). Fruits eaten, after leaching; bark used in decoctions for toothache, loose teeth; salve of pulverized fruits applied to hemorrhoids; fruits used in preparation of fish poison. (1)" ^(Bocek)

Aesculus chinensis

"In Chinese herbal medicine, the seed of a related plant, A. chinensis, is used to treat malnutrition and other digestive difficulties at a dose of 3 to 9 grams in decoction. Japanese herbal medicine prescribes the seed of A. turbinata, another related plant, to treat digestive difficulties and promote absorption.⁸⁰" ^{[Boik NCCT]}

"Aesculus chinensis is native to China and is distributed mainly in the Hebei, Henan, and Shanxi provinces. The fruits of this species usually are smooth subglobose, truncate to slightly impressed at the apex, thickly walled with smaller seeds having proportionately larger hilum. The surface of its petiolulate leaflets is glabrous or sparingly pilose [2]. Its seeds, called "Sha Luo Zhi" in traditional Chinese medicine, have long been used as a stomachic and analgesic in the treatment of distention and pain in chest and abdomen, ma- laria, and dysentery [10]. Additionally, the tablets made from the seeds are used for treating heart diseases [11]." ^{(Shiyou Li et al.,2010)}

Aesculus indica

Aesculus indica (Colebr. Ex. Cambess.) Hook. "Ethnomedicinal uses: Oil from the seeds is externally applied against rheumatism. Seeds are given to horses in colic pain." ^{(Kumar et al.,2009)}

"Aesculus indica, known as Indian horse chestnut, is found on mountain slopes or in moist and shady valleys in the northwestern Himalayan forests. It is distributed from Nepal northwestward into the State of Kashmir in north India, and across the Indus River to West Pakistan and to northeastern Afghanistan. Its fruits are reddish brown, smooth ovoids. The petiolulate leaflets of this species are submembranacous with finely serrate margin [2]. In some parts of Himachal Pradesh, the seeds are dried and grounded into flour, called tattwakhar. This flour is bitter and used for making halwa, which is taken as phalahar (non-cereal food) during fasts; the leaves are used as a fodder for cattle. India horse chestnut also has medicinal properties for animals and human beings. The fruits are given to horses suffering from colic. The oil extracted from the seeds is used to cure rheumatism [21]." ^{(Shiyou Li et al.,2010)}

Aesculus pavia

"Recently, a new prenylated coumarin with antifungal activity, pavietin (161), was isolated and identified from the leaves of A. pavia [67]." ^{(Shiyou Li et al.,2010)}

Aesculus turbinata

"Aesculus turbinata, known as Japanese horse chestnut, is native only to Japan. It has been found on the island of Hokkaido and the central and northern parts of Honshu. This species has been widely cultured in England, the United States, and China. This species is similar to A. hippocastanum, differently mainly in the slightly smaller flowers, warty fruit surface, and in the larger leaves, which are glaucescent beneath and have more regularly crenate-serrate margins [2]. The seeds of the Japanese horse chestnut have been used as an emergency provision since ancient times and utilized traditionally in Japan as a confectionery ingredient in rice cakes and rice balls [19]. Its seed extract in combination with spirits have also been used as a folk medicine for the treatment of bruises and sprains in some regions of Japan [20]." ^{(Shiyou Li et al.,2010)}

"The seeds of A. turbinata also contain rich triterpenoid saponins as the main components. A number of investigations on the seeds of Japanese horse chestnut resulted in the isolation and structure determination of 16 polyhydroxylated triterpenoid saponins." ^{(Shiyou Li et al.,2010)}

References

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• (baraldi2007) Baraldi, Cecilia, et al. "Chemical composition and characterisation of seeds from two varieties (pure and hybrid) of Aesculus hippocastanum." Food chemistry 104.1 (2007): 229-236.
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• Duke - http://www.ars-grin.gov/cgi-bin/duke/ethnobot.pl?Aesculus%20hippocastanum Accessed Dec 23, 2014
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• E-Flora BC, E-flora - http://ibis.geog.ubc.ca/biodiversity, Electronic Atlas of the Flora of British Columbia, Klinkenberg, Brian. (Editor) 2013. E-Flora BC: Electronic Atlas of the Flora of British Columbia (eflora.bc.ca). Lab for Advanced Spatial Analysis, Department of Geography, University of British Columbia, Vancouver.
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• Herbs2000 - Aesculus hippocastanum, Accessed May 18, 2014
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• (Hubner,1999) Hübner, Gabriele, Victor Wray, and Adolf Nahrstedt. "Flavonol oligosaccharides from the seeds of Aesculus hippocastanum." Planta Medica 65.07 (1999): 636-642.
• (Irving H. Isenberg) Isenberg, Irving H. "Papermaking fibers." Economic Botany 10.2 (1956): 176-193.
• Jepson2012 - William J. Stone, 2012. Aesculus, in Jepson Flora Project (eds.), Jepson eFlora, http://ucjeps.berkeley.edu/cgi-bin/get_IJM.pl?tid=12026, accessed on Mar 6 2014, January 10, 2019.
• (kapusta2007) Kapusta, Ireneusz, et al. "Flavonoids in horse chestnut (Aesculus hippocastanum) seeds and powdered waste water byproducts." Journal of agricultural and food chemistry 55.21 (2007): 8485-8490.
• (kim1994) Kim, Nicholas D., and Jack E. Fergusson. "Seasonal variations in the concentrations of cadmium, copper, lead and zinc in leaves of the horse chesnut (Aesculus hippocastanum L.)." Environmental Pollution 86.1 (1994): 89-97.
• (konoshima1986) Konoshima, Takao, and Kuo-Hsiung Lee. "Antitumor agents, 82. Cytotoxic sapogenols from Aesculus hippocastanum." Journal of natural products 49.4 (1986): 650-656.
• (Kumar et al.,2009) Kumar, Mahesh, Yash Paul, and V. K. Anand. "An ethnobotanical study of medicinal plants used by the locals in Kishtwar, Jammu and Kashmir, India." Ethnobotanical leaflets 2009.10 (2009): 5.
• (matsuda1997) Matsuda, Hisashi, et al. "Antiinflammatory effects of escins Ia, Ib, IIa, and IIb from horse chestnut, the seeds of Aesculus hippocastanum L." Bioorganic & Medicinal Chemistry Letters 7.13 (1997): 1611-1616.
• (Ortiz,GTR) Ortiz, Beverly R. "Contemporary California Indians, oaks and sudden oak death (Phytophthora ramorum)." In: Merenlender, Adina; McCreary, Douglas; Purcell, Kathryn L., tech. eds. 2008. Proceedings of the sixth California oak symposium: today's challenges, tomorrow's opportunities. Gen. Tech. Rep. PSW-GTR-217. Albany, CA: US Department of Agriculture, Forest Service, Pacific Southwest Research Station: pp. 39-56. Vol. 217. 2008.
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• (Pieroni, Andrea, et al.) Pieroni, Andrea, et al. "Ethnopharmacognostic survey on the natural ingredients used in folk cosmetics, cosmeceuticals and remedies for healing skin diseases in the inland Marches, Central-Eastern Italy." Journal of Ethnopharmacology 91.2-3 (2004): 331-344.
• PFAF - Aesculus hippocastanum, Plants For A Future, http://www.pfaf.org/user/Plant.aspx?LatinName=Aesculus+hippocastanum, Accessed Jan 12, 2015
• (Podolak et al,2010) Podolak, Irma, Agnieszka Galanty, and Danuta Sobolewska. "Saponins as cytotoxic agents: a review." Phytochemistry Reviews 9.3 (2010): 425-474.
• (Redzic,2007) S Redžić, Sulejman. "The ecological aspect of ethnobotany and ethnopharmacology of population in Bosnia and Herzegovina." Collegium antropologicum 31.3 (2007): 869-890.
• (Saric-kundalic et al., 2010) Šarić-Kundalić, Broza, et al. "Ethnobotanical study on medicinal use of wild and cultivated plants in middle, south and west Bosnia and Herzegovina." Journal of ethnopharmacology 131.1 (2010): 33-55.
• (Shiyou Li et al.,2010) Li, Shiyou, Xiao-Yuan Lian, and Zhizhen Zhang. "An overview of genus Aesculus L.: ethnobotany, phytochemistry, and pharmacological Activities." (2010).
• Wiki - Aesculus hippocastanum, Wikipedia.org, https://en.wikipedia.org/wiki/Aesculus_hippocastanum
  • [3] Rushforth, K. (1999). Trees of Britain and Europe. Collins ISBN 0-00-220013-9.
  • [5] Euro+Med Plantbase Project: Aesculus hippocastanum Archived September 28, 2007, at the Wayback Machine.
• (wilkinson1999) Wilkinson, J. A., and A. M. G. Brown. "Horse Chestnut–Aesculus hippocastanum: Potential applications in cosmetic skin‐care products." International journal of cosmetic science 21.6 (1999): 437-447.
• (Yilmaz,2006) Yilmaz, Ruya, et al. "Use of Aesculus hippocastanum L. as a biomonitor of heavy metal pollution." Pak. J. Bot 38.5 (2006): 1519-1527.

Image References

• 1, Chestnut inflorescences (Aésculus hippocástanum), Ввласенко, CC BY-SA 3.0 , via Wikimedia Commons
• 2, Fruit de marronnier (Aesculus hippocastanum), Gzen92, CC BY-SA 4.0 , via Wikimedia Commons

Journals of Interest

• Yılmaz, R., S. Sakcalı, C. Yarcı, A. Aksoy and M. Ozturk, 2006. Use of Aesculus hippocastanum L. as a biomonitor of heavy metal pollution. Pak. J. Bot., 38(5): 1519-1527.
• Kim, N.D., Fergusson, J.E., 1994. Seasonal variations in the concentrations of cadmium, copper, lead and zinc in leaves of the horse chesnut (Aesculus hip- pocastanum L.). Environ. Pollut. 86, 89–97.
• De La Montana Miguelez, J., Miguez Bernardez, M., & Garcia Queijeiro, J. M. (2004). Composition of varieties of chestnuts from Galicia (Spain). Food Chemistry, 84, 401–404.
• Deli, J., Matus, Z., & Toth, G. (2000). Comparative study on the carotenoid composition in the buds and flowers of different Aesculus species. Chromatographia, 51, 179–182.
• Deli, J., Molnar, P., Matus, Z., Toth, G., Steck, A., Niggli, U. A., et al. (1998). Aesculaxanthin, a new carotenoid isolated from pollens of Aesculus hippocastanum. Helvetica Chimica Acta, 81, 1815–1820.
• Facino RM, Carini M, Stefani R, Aldini G, Saibene L: Anti-elastase and anti-hyaluronidase activities of saponins and sapogenins from Hedera helix, Aesculus hippocastanum, and Ruscus aculeatus: factors contributing to their efficacy in the treat- ment of venous insufficiency. Arch Pharm (Weinheim) 1995, 328:720-4.
• Konoshima, T., & Lee, K. H. (1986). Antitumor agents, 82. Cytotoxic sapogenols from Aesculus hippocastanum. Journal of Natural Prod- ucts, 49, 650–656.
• Wei, F., Ma, L. Y., Cheng, X. L., Lin, R. C., Jin, W. T., Khan, I. A., et al. (2005). Preparative HPLC for purification of four isomeric bioactive saponins from the seeds of Aesculus chinensis. Journal of Liquid Chromatography and Related Technologies, 28, 763–773.
• Yang, X. W., Zhao, J., Cui, Y. X., Liu, X. H., Ma, C. M., Hattori, M., et al. (1999). Anti-HIV-1 protease triterpenoid saponins from the seeds of Aesculus chinensis. Journal of Natural Products, 62, 1510–1513.
• Yoshikawa, M., Murakami, T., Matsuda, H., Yamahara, J., Murakami, N., & Kitagawa, I. (1996). Bioactive saponins and glycosides. 3. Horse chestnut. Chemical & Pharmaceutical Bulletin, 44, 1454–1464.
• Newell, E.A. (1991) Direct and delayed costs of reproduction in Aesculus californica. Journal of Ecology 79, 365–378.
• Zhang Z, Li S (2007) Cytotoxic triterpenoid saponins from the fruits of Aesculus pavia L. Phytochemistry 68:2075–2086

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