Flavonol glycosides in the petal of Rosa species as chemotaxonomic markers Sarangowa, Ochir Kanazawa, Tsutomu Nishizawa, Makoto Myoda, Takao Bai, Changxi Yamagishi, Takashi Phytochemistry 2014 2014-11-30 107 61 68 6WWCX 74632 L. [483,569,599,619] Magnoliopsida Rosaceae Rosa Plantae Rosales 1 62 Tracheophyta species gallica  The 50% EtOH extracts of the petals of a hybrid of  R. gallicacollected in the Xinjiang provincein Chinawere continuously partitioned with a H 2O–MeOH–CHCl 3system ,and the upper layer was fractionated by column chromatography on MCI gel CHP-20P followed by chromatography on Sephadex LH-20. The fractions containing flavonol glycosides were further purified using a reversed phase gel (Wakosil 40 C18) to give compounds 1–13. Compounds 1, 2, 3, 5and 9were isolated as the major flavonol glycosides in the petals of  R. gallica, and were identified as quercetin 3- O-(  200- O - β-D- glucosyl)- β-D- galactoside ( 1), quercetin 3- O-sophoroside ( 2), kaempferol 3- O-sophoroside ( 3), quercetin 3- O - β-D- glucoside ( 5) and kaempferol 3- O - β-D- glucoside ( 9), respectively, as previously described ( Sarangowa et al., 2010) and by comparison of their 1H and 13C NMR data and MS data with literature values ( Iwashina et al., 2005; Schliemann et al., 2006; Ross et al., 2005). The 1H NMR spectra of 4, 10and 11indicated the presence of quercetin as an aglycone. The 13C NMR spectrum of compound 4showed the presence of a hexose, and the spectra of 10and 11showed the presence of a pentose. Comparing the 1H and 13C NMR data and MS data with literature data ( Lin and Lin, 1999; Nowak and Wolbis, 2002; Park et al., 2012; Tanaka et al., 1981; Zhu et al., 2013), the structures of 4, 10and 11were identified as quercetin 3- O - β-D- galactoside ( 4), quercetin 3- O - β-D- xylopyranoside ( 10) and quercetin 3- O - α-L- arabinofuranoside ( 11), respectively. The positions of the sugar moiety on quercetin were confirmed by analysis of HMBC spectra. The C 1–H of the sugar moiety gave a cross peak with the C 3position of quercetin. The 1H NMR spectra of compounds 8, 12and 13indicated presence of kaempferol as the aglycone ( Xiao et al., 2006; Zhu et al., 2013). The 13C NMR spectrum of 8showed the presence of a hexose, and the spectra of 12and 13showed the presence of a pentose. Comparing the 1H and 13C NMR data and MS data of these compounds with literature values ( Nowak and Wolbis, 2002; Alaniya et al., 2012; Park et al., 2012), the structures of 8, 12and 13were characterized as kaempferol 3- O - β-D- galactoside ( 8), kaempferol 3- O - α-L- arabinofuranoside ( 12), and kaempferol 3- O - α-D- rhamnoside ( 13). Quercetin 3- O-(  200-O-galloyl)- β-D- glucoside ( 6) and kaempferol 3- O-(  200-O-galloyl)- β-D- glucoside ( 7) were also isolated from  R. gallicaas detailed in our previous report ( Sarangowa et al., 2013) ( Fig. 1).   2.2. Quantitative determination of flavonol glycosides in sections   Gallicanae, Cinnamomeae, Caninaeand Synstylae A method using ultra-performance liquid chromatography (UPLC) equipped with a photodiode array (PDA) detector was optimized for quantitative determination of flavonol glycosides in the 50% EtOH extract of the petal of 31  Rosaspecies( Table 1). The monitoring wavelength was set at 350 nm, which was the absorption maximum for glycosides of quercetin and kaempferol. Chromatography peaks of quercetin and kaempferol glycosides were identified from their retention times and UV spectra monitored by PDA detector. The content of flavonol glycosides was quantitatively analyzed by UPLC. Typical chromatograms of hybrids of  R. gallicaand  R. rugosaare shown in Fig. 2. Seven glycosides of quercetin ( 1, 2, 4, 5, 6, 10and 11) and six glycosides of kaempferol ( 3, 7, 8, 9, 12and 13) were detected in the chromatogram of the hybrid of  R. gallica. However, in the chromatogram of  R. rugosa, 3- O-(  200-O - β-D- glycosyl)- β-D- xylosides of quercetin ( 14) and kaempferol ( 15) were detected along with 2and 3, while other glycosides were not detected. The content of fifteen glycosides of quercetin and kaempferol are shown in Table 2. Fifteen flavonol glycosides were identified in this experiment; however, all of the flavonol glycosides were not detected in all samples. The total content of these glycosides was highest in the petals of  Rosaspeciesin section  Gallicanae( 28.68–119.10 mg/g), followed by section  Caninae(30.78 and 41.52 mg/g) and section  Synstylae(30.49 and 48.90 mg/g). The glycoside content in the petals of  Rosaspeciesin Section  Cinnamomeaewas lower than the other sections ( 0.48–7.5 mg/g). Flavonol glycosides including a hexose ( 4, 5, 8, 9and 13) and their galloyl derivatives ( 6and 7), and those with a pentose ( 10, 11and 12) were detected only in section  Gallicanae, except for  Rosa acicularis(Ci05),  Rosa davurica(Ci08) and  R. rugosa× Rosa rubrifoia(Ci02). On the other hand, the  Rosaspeciesin section  Cinnamomeaemainly contained glycosides with a disaccharide ( 1, 2, 3, 14and 15). Sophorosides of quercetin and/or kaempferol ( 2and 3, respectively) were detected in almost all of the samples, and two patterns involving the presence and content of these glycosides along with the content of 1were recognized. Samples belong to pattern 1 contained relatively large amount of 1, 2and 3, and those belonging to pattern 2 contained lower amounts of these compounds (or were not detected). Compounds 14and 15were detected only in section  Cinnamomeae.    3. Discussion   3.1. Flavonol glycosides in R. gallica  Xiao et al. (2006)reported the isolation of glycosides of kaempferol and quercetin such as 1, 3, 4, 9, 11and 12from ‘‘  R. rugosa’’ collected in the Xinjiang province, China. In the present study, samples of  Rosaplants were collected at the same location for analysis. As reported previously, the roses used as ‘‘Mei-gui’’ or ‘‘Kizil gul’’ in Xinjiang provinceare not  R. rugosa, but are  R. gallicaand its hybrids. In this paper, glucosides 4, 9, 11and 12were not detected in 6 samples of  R. rugosacollected in Japan, Koreaand China, and 1, 3, 4, 9, 11and 12were detected in all of the samples collected in Xinjiang province. Therefore, the plants collected by Xiao et al. (2006)were supposed to be hybrids of  R. gallica. In this study, additional glycosides of quercetin and kaempferol including 2, 5, 8, 10and 13were isolated for the first time from hybrids of  R. gallica.