**2.12.** *Smilax lanceaefolia*

The root of *S*. *lanceifolia* prepared as a decoction is used in traditional Indian medicine to soothe stomach pain and rheumatism. The boiled extract is used to expel gallbladder and kidney stones. It was also found that the aqueous extract of *S*. *lanceifolia* contains compounds with a high affinity for binding proteins, specifically by active sites joining reverse transcriptase. Therefore, it can be used to inhibit proliferation of retroviruses-agents in viral diseases such as AIDS and T-cell leukemia [51]. Laintojam and Kongbrailatpam performed a study on the chemical constituents and antioxidant activiy of *S*. *lanceifolia* roots extracts [7]. The extracts were obtained from petroleum ether, chloroform and methanol, successively. Antioxidant

Phenolic Compounds in Genus *Smilax* (Sarsaparilla) http://dx.doi.org/10.5772/66896 249

(**26**), (−)epicatechin (**27**) and cinchonain 1b (**60**)); 2 flavanones (luteolin (**62**) and apigenin (**63**)); two flavonols (quercetin (**7**) and myricetin (**64**)); 1 chalcone, kukulkanin B (**65**); 3 stilbenes (piceid (**38**), piceatannol (**39**), and resveratrol (**40**)); 6 phenylpropanoids (5-*O*caffeoylshikimic acid (**45**), caffeic acid (**66**), 3-*O*-*p*-coumaroylshikimic acid (**67**), smiglycerol (**68**), juncusyl ester B (**69**) and 1-*O*-p-coumarylglycerol (**70**), **Figure 14**). It is noteworthy that in this study no smiglasides were detected, despite an intensive separation of phenolic compounds having conducted. One explanation for these results is that possibly the high temperatures used for

*S*. *glycyphylla* is a plant endemic to Australia and its leaves and fruits have a sweet taste like honey grass. The sweet principle of *S*. *glycyphylla* is called glycyphyllin A (**71**) and identified as a phenol compound with a structure of dihydrochalcone, phloretin2′αLrhamnose [48]. Cox *et al*. evaluated aqueous extracts of leaves and stems of *S. glyciphylla* [49]. The methods used to evaluate antioxidant activity were lipid peroxidation using thiobarbituric acid reactive substances (TBARS), superoxide quenching by coupling superoxide generation to the reduction of nitroblue tetrazolium (NBT), inhibition of deoxyribose-driven fenton degradation and total radical-antioxidant potential (TRAP) using free radicals derived from ABTS (2,2′azinobis(3ethylbenzothiazoline 6sulphonate). The results showed that *S*. *glycyphylla* extract inhibited deoxyribose degradation. The total radical-antioxidant potential was seven times that the trolox a water-soluble analog of vitamin E with a high antioxidant activity.

Huang *et al*. performed a study of phenolic profile and antioxidant activity of *S*. *glycyphylla* leaves [50]. The leaves of *S*. *glycyphylla* previously dried and blended were extracted with 80% aqueous ethanol. Then, the extract was concentrated and partitioned with hexane and butanol, consecutively. The butanol fraction was subjected to chromatographic separation. The separation produced eight phenolic compounds such as the glycyphyllin A previously mentioned, two new dihydrochalcones (glycyphyllin B (**72**) and C (**73**)) and five flavonoids (catechin (**26**), (2*R*,3*R*)-dihydrokaempferol-3-*O*-*β*-D-glucopyranoside (**57**), kaempferol-3-*Oβ*-D-glucopyranoside (**74**), quercetin-3-*O*-*β*-D-glucopyranoside (**75**) and kaempferol-3-*O*-*β*neohesperidoside (**76**), **Figures 14** and **15**). The antioxidant activity of pure compounds was evaluated using ferric reducing ability of plasma (FRAP) and DPPH• radical scavenging. The flavonoids showed a good antioxidant activity; contrary to this, the dihydrochalcones showed

The root of *S*. *lanceifolia* prepared as a decoction is used in traditional Indian medicine to soothe stomach pain and rheumatism. The boiled extract is used to expel gallbladder and kidney stones. It was also found that the aqueous extract of *S*. *lanceifolia* contains compounds with a high affinity for binding proteins, specifically by active sites joining reverse transcriptase. Therefore, it can be used to inhibit proliferation of retroviruses-agents in viral diseases such as AIDS and T-cell leukemia [51]. Laintojam and Kongbrailatpam performed a study on the chemical constituents and antioxidant activiy of *S*. *lanceifolia* roots extracts [7]. The extracts were obtained from petroleum ether, chloroform and methanol, successively. Antioxidant

extraction caused smiglasides degradation.

248 Phenolic Compounds - Natural Sources, Importance and Applications

**2.11.** *Smilax glycyphylla*

weak antioxidant activity [50].

**2.12.** *Smilax lanceaefolia*

**Figure 14.** Flavonoids and phenylpropanoids isolated from *S*. *glabra* rhizome.

**Figure 15.** Phenolic compounds isolated from 80% ethanol extract of *S*. *glycyphylla* leaves.

activity of the extracts was evaluated by DPPH• radical scavenging and it was found that the methanol extract had the principal antiradical activity attributed to phenolic compounds. The only compound isolated from methanol extract was flavanonol glycoside, quercitrin (**77**, **Figure 16**) [7].

### **2.13.** *Smilax riparia*

The roots and rhizomes of *S. riparia*, commonly called "Niu-Wei-Cai" in China, are used in traditional Chinese medicine as diuretics, treatments for inflammation and cancer [52], and in some cases as food [53]. Sun *et al*. isolated three phenylpropanoid glycosides (the smilasides M (**78**) and N (7**9**), and 2′,6′diacetyl3,6diferuloylsucrose (**80**), **Figure 16**) from a 95% aqueous ethanol extract of *S*. *riparia* roots and rhizomes. The concentrated extract was suspended

**Figure 16.** Flavanonol glycoside from methanol extract of *S*. *lanceifolia* roots*.*

in water and subjected to D101 macroporous resin column chromatography and eluted with water and 30% and 70% ethanol, successively. The fraction eluted with 70% ethanol was suspended in water and partitioned with chloroform, ethyl acetate and butanol. The ethyl acetate fraction was chromatographed over a silica gel column, followed by thin-layer chromatography (TLC) and finally, the fractions were subjected to C18 reversedphase silica gel column to obtain the afore-mentioned compounds, [54].

Wang *et al*. isolated five phenylpropanoids with a sucrose core (helonioside B (**18**), smiglaside A (**51**), smiglaside B (**52**), and smilaside P (**81**), **Figures 5**, **13** and **17**) [55]. The phenylpropanoid compounds were obtained from a 95% aqueous ethanol extract of *S*. *riparia* roots and rhizomes. The extract was subjected to macroporous resin HPD600 and eluted with 95% aqueous ethanol and ethyl acetate. Subsequently, the ethyl acetate fraction was subjected to silica gel column chromatography, and from this fraction were obtained the phenylpropanoids compounds. The five compounds were subjected to cytotoxicity test against human promyelocytic leukemia (HL-60), human hepatocellular carcinoma (SMMC-7721), human lung cancer (A-549), human breast cancer (MCF-7) and human colon cancer (SW480) using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS). The five compounds were evaluated for anticancer activity. Antioxidant activity was also evaluated using DPPH• radical scavenging activity. The results showed the phenylpropanoid compounds with three feruloyl and acetyl groups exhibited the primary antitumoral and antioxidant activities. The proposed explanation of these results was that the feruloyl and


**Figure 17.** Phenylpropanoids from 95% ethanol extract of *S*. *riparia* roots and rhizomes.

activity of the extracts was evaluated by DPPH• radical scavenging and it was found that the methanol extract had the principal antiradical activity attributed to phenolic compounds. The only compound isolated from methanol extract was flavanonol glycoside, quercitrin (**77**,

**Figure 15.** Phenolic compounds isolated from 80% ethanol extract of *S*. *glycyphylla* leaves.

250 Phenolic Compounds - Natural Sources, Importance and Applications

**Figure 16.** Flavanonol glycoside from methanol extract of *S*. *lanceifolia* roots*.*

The roots and rhizomes of *S. riparia*, commonly called "Niu-Wei-Cai" in China, are used in traditional Chinese medicine as diuretics, treatments for inflammation and cancer [52], and in some cases as food [53]. Sun *et al*. isolated three phenylpropanoid glycosides (the smilasides M (**78**) and N (7**9**), and 2′,6′diacetyl3,6diferuloylsucrose (**80**), **Figure 16**) from a 95% aqueous ethanol extract of *S*. *riparia* roots and rhizomes. The concentrated extract was suspended

**Figure 16**) [7].

**2.13.** *Smilax riparia*

acetyl groups confer a minor polarity to the compounds with the most activity and are key to inducing antitumoral activity [55].

### **2.14.** *Smilax scobinicaulis*

The roots of *S. scobinicaulis*, also called "Hei Ci Ba Quia" in Chinese, are used in traditional Chinese medicine for the treatment of arthritis, gout and inflammatory diseases [10]. Zhang *et al*. studied the chemical composition of *S*. *scobinicaulis*. These researchers first obtained a 95% aqueous ethanol extract from *S*. *scobinicaulis* roots and rhizomes. The extract was concentrated, suspended in water and partitioned with petroleum ether, ethyl acetate and butanol, successively. The ethyl acetate fraction was subjected to column chromatography. The purification was done to isolate and characterize two new flavones (7,3′,5′trihydroxy5,6,4′ trimethoxyflavone (**82**) and 7hydroxy5,6,3′,5′pentamethoxyflavone (**83**), **Figure 18**). The new flavones were evaluated for cell proliferation and viability assay against human breast adenocarcinoma (MCF-7) and human lung carcinoma (H520). The results showed a weak activity for **82**, but 7hydroxy5,6,3′,5′pentamethoxyflavone, **83** was inactive [10].

### **2.15.** *Smilax sebeana*

*S. sebeana* is used in traditional Japanese and Chinese medicine to treat syphilis, arthritis and gout [11]. Ao *et al*. evaluated antioxidant activity in isolated phenolic compounds from methanol extracts of *S*. *sebeana* rhizomes and roots [11]. The methanol extract was obtained from maceration of fresh rhizomes and roots with methanol at room temperature. The methanol extract was concentrated, suspended in water and partitioned with hexane, ethyl acetate and butanol, successively. The ethyl acetate fraction was purified in two steps. First, it was subjected to column chromatography packed with sephadex LH20. Some of the fractions collected were selected to evaluate by HPLC to evaluate their components. Finally, the fractions with similar composition were pooled and purified by preparative HPLC. Also, the total phenol content and antioxidant activity expressed by DPPH radical scavenging were evaluated for methanol, ethyl acetate and butanol fractions. The major content of total phenols was

**Figure 18.** Flavones from 95% ethanol extract of *S*. *scobinicaulis* roots.

found in the ethyl acetate fraction, 238.5 mg catechin/g extract. This fraction also showed the principal DPPH• radical scavenging activity, IC50 of approximately 10.4 μg/mL. The compounds isolated from the ethyl acetate fraction were one phenylpropanoid (chlorogenic acid (**84**), **Figure 19**) and three cinchonain (1b (**61**), 1a (**85**) and 11a (**86**), **Figures 14** and **19**) [11].
