G. A. Zou

Urinary metabonomics study of anti-depressive effect of Chaihu-Shu-Gan-San on an experimental model of depression induced by chronic variable stress in rats

Chaihu-Shu-Gan-San (CSGS), a traditional Chinese medicine (TCM) formula, has been effectively used for the treatment of depression in clinic. However, studies of its anti-depressive mechanism are challenging, accounted for the complex pathophysiology of depression, and complexity of CSGS with multiple constituents acting on different metabolic pathways. The variations of endogenous metabolites in rat model of depression after administration of CSGS may offer deeper insights into the anti-depressive effect and mechanism of CSGS. In this study, metabonomics based on ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) was used to profile the metabolic fingerprints of urine obtained from chronic variable stress (CVS)-induced depression model in rats with and without CSGS treatment. Through partial least squares-discriminate analysis, it was observed that metabolic perturbations induced by chronic variable stress were restored in a time-dependent pattern after treatment with CSGS. Metabolites with significant changes induced by CVS, including 3-O-methyldopa (1), pantothenic acid (2), kynurenic acid (3), xanthurenic acid (4), 2,8-dihydroxyquinoline glucuronide (5), 5-hydroxy-6-methoxyindole glucurnoide (8), l-phenylalanyl-l-hydroxyproline (9), indole-3-carboxylic acid (10), proline (11), and the unidentified metabolites (6, 2.11min_m/z 217.0940; 7, 2.11min_m/z 144.0799), were characterized as potential biomarkers involved in the pathogenesis of depression. The derivations of all those biomarkers can be regulated by CSGS treatment except indole-3-carboxylic acid (10), which suggested that the therapeutic effect of CSGS on depression may involve in regulating the dysfunctions of energy metabolism, tryptophan metabolism, bone loss and liver detoxification. This study indicated that the rapid and noninvasive urinary metabonomics approach may be a powerful tool to study the efficacy and mechanism of complex TCM prescriptions.

Online identification of the antioxidant constituents of traditional Chinese medicine formula Chaihu-Shu-Gan-San by LC–LTQ-Orbitrap mass spectrometry and microplate spectrophotometer

Chaihu-Shu-Gan-San (CSGS), a traditional Chinese medicine (TCM) formula containing seven herbal medicines, has been used in treatment of gastritis, peptic ulcer, irritable bowel syndrome and depression clinically. However, the chemical constituents in CSGS had not been studied so far. To quickly identify the chemical constituents of CSGS and to understand the chemical profiles related to antioxidant activity of CSGS, liquid chromatography coupled with electrospray ionization hybrid linear trap quadrupole orbitrap (LC-LTQ-Orbitrap) mass spectrometry has been applied for online identification of chemical constituents in complex system, meanwhile, antioxidant profile of CSGS was investigated by the fraction collecting and microplate reading system. As a result, 33 chemical constituents in CSGS were identified. Among them, 13 components could be detected both in positive and in negative ion modes, 20 constituents were determined only in positive ion mode and 2 components were only detected in negative ion mode. Meanwhile, the potential antioxidant profile of CSGS was also characterized by combination of 96-well plate collection of elutes from HPLC analysis and microplate spectrophotometer, in which the scavenging activities of free radical produced by DPPH of each fraction could be directly investigated by the analysis of microplate reader. This study quickly screened the contribution of CSGS fractions to the antioxidant activity and online identified the corresponding active constituents. The results indicated that the combination of LC-MS(n) and 96-well plate assay system established in this paper would be a useful strategy for correlating the chemical profile of TCMs with their bioactivities without isolation and purification.

Flavonoids from the Stems of Croton caudatus Geisel. var. tomentosus Hook

A new flavone, named crotoncaudatin (1), was isolated from the stems of Croton caudatus Geisel. var. tomentosus Hook., together with nine known analogues: 3,5,6,7,8,3′,4′-heptamethoxyflavone (2), tangeretin (3), nobiletin (4), 5,6,7,4′-tetramethoxy-flavone (5), sinensetin (6), kaempferol (7), tiliroside (8), kaempferol-3-O-rutinoside (9) and rutin (10). The structures of the above compounds were established by a combination of spectroscopic methods, including HR-ESI-MS, 1H-NMR, 13C-NMR, HMQC and HMBC spectra. All compounds were isolated from and identified in this species for the first time and compounds 1-6 are new for the genus Croton.

Pyrrole alkaloids from Alhagi sparsifolia

The novel pyrrole alkaloid alhagifoline A (1) together with the two known analogs pyrrolezanthine (2) and pyrrolezanthine-6-methyl ether (3) were isolated from the aerial part of Alhagi sparsifolia. Their structures were established based on spectral (HR-ESI-MS, 1 H and 13C NMR, 1 H–1 H COSY, HSQC, HMBC) data. Compounds 2 and 3 were isolated from the genus Alhagi for the first time.

Chemical constituents from seeds of Lactuca sativa

A new flavonol glycoside with a rare structure type, lactucasativoside A (1), was isolated from the seeds of Lactuca sativa, together with three known compounds, japonicin A (2), isoquercitrin (3), and caffeic acid (4). Their structures were established on the basis of extensive spectroscopic analysis. Compound 2 was obtained from the genus Lactuca for the first time.

flavonoids from croton laevigatus (a)

Chinese National Program for Treatment of Infectious Diseases;Natural Sciences of China;National Specialized Project for Innovations of New Drugs

Three new diterpenoids from Coleus forskohlii Briq

Three new diterpenoids, forskolin G(2), forskolin H(3), forskolin I(4), were isolated from the whole plant of the Coleus forskohlii Briq., and their structures were elucidated as 1α,6β-diacetoxy-8,13-epoxylabd-14-en-11-one, 1α-hydroxy-6β,7β-diacetoxy-8,13-epoxylabd-14-en-11-one, and 1α,9α-dihydroxy-6β,7α-diacetoxy-8,13-epoxylabd-14-en-11-one on the basis of spectral data.

Chemical Composition of Alhagi sparsifolia Flowers

Alhagi sparsifolia (Leguminosae) is a perennial herb or semi-bush that is broadly distributed in the flora of Central Asia, i.e., Tajikistan, Kazakhstan, Uzbekistan, Turkmenistan, and Kyrgyzstan [1]. It grows along river banks and in arid zones of the provinces Gansu, Inner Mongolia, and Qinghai and Xinjiang–Uyghur Autonomous Region (XUAR) of China [2]. It is used in traditional Uyghur medicine to treat fever, rheumatism, diarrhea, stomach pain, headache, toothache, and cancer [3]. Flowers are usually used as a tincture for use by local residents. Previously, catechins, proanthocyanidins, coumarins, flavonoids, steroids, and alkaloids were isolated from the plant [4, 5]. Dried flowers (7 kg) were collected in XUAR and extracted by refluxing EtOH (75%) in a flask at 85°C for 3 h. The condensed extract (497.7 g) obtained after solvent distillation was suspended in H2O and worked up sequentially with petroleum ether, EtOAc, and n-BuOH. The EtOAc fraction (57.7 g) was chromatographed over a column of silica gel followed by gradient elution by CH2Cl2–MeOH (1:0 0:1) and purification over a column of Sephadex LH-20 to afford compounds 1–15. The structures of 1–15 were determined using spectral data (1D and 2D NMR and mass spectroscopy) and comparisons of the analyses with those published [6–20]. Genistein (2), isovanillic acid (3), syringetin (4), gentisic acid (10), and genistin (11) were isolated for the first time from the genus Alhagi; heptacosan-1-ol (1) and gallic acid (14) were isolated for the first time from this plant. Heptacosan-1-ol (1), 80.6 mg, white amorphous powder, C27H56O, mp 82–83 C. ESI-MS, m/z: 395.3 [M – H] –. 1Í NMR spectrum (400 MHz, CDCl3, , ppm, J/Hz): 3.64 (3H, t, J = 6.6, H-1), 1.57 (2H, dt, J = 13.7, 6.7, H-2), 1.51–0.96 (48H, m, H-3–26), 0.88 (3H, t, J = 6.7, H-27) [6]. Genistein (2), 14.9 mg, pale-yellow amorphous powder, Cl5Hl0O5, mp 296–298 C. ESI-MS, m/z: 269.7 [M – H] –. 1Í NMR spectrum (400 MHz, CD3OD, , ppm, J/Hz): 8.07 (1H, s, H-2), 7.39 (2H, d, J = 8.6, H-2 , 6 ), 6.86 (2H, d, J = 8.6, H-3 , 5 ), 6.36 (1H, br.s, H-8), 6.24 (1H, br.s, H-6) [7]. Isovanillic acid (3), 3.8 mg, white needle-like crystals, Cl5Hl0O5, mp 296–298 C. ESI-MS, m/z: 269.7 [M – H] –. 1Í NMR spectrum (400 MHz, CD3OD, , ppm, J/Hz): 8.07 (1H, s, H-2), 7.39 (2H, d, J = 8.6, H-2 , 6 ), 6.86 (2H, d, J = 8.6, H-3 , 5 ), 6.36 (1H, br.s, H-8), 6.24 (1H, br.s, H-6) [7]. Syringetin (4), 8.6 mg, pale-yellow amorphous powder, C17H14O8, mp 224–226 C. ESI-MS, m/z: 345.5 [M – H] –. 1Í NMR spectrum (400 MHz, DMSO-d6, , ppm, J/Hz): 12.44 (1H, s, 5-OH), 10.75 (1H, s, 7-OH), 9.47 (1H, s, 3-OH), 9.13 (1H, s, 4 -OH), 7.51 (2H, s, H-2 , 6 ), 6.52 (1H, d, J = 2.0, H-8), 6.20 (1H, d, J = 2.0, H-6), 3.84 (6H, s, 3 , 5 -OCH3) [9]. Isorhamnetin (5), 6.1 mg, yellow amorphous powder, C16H12O7, mp 303–305 C. ESI-MS, m/z: 315.9 [M – H] –. 1Í NMR spectrum (400 MHz, DMSO-d6, , ppm, J/Hz): 12.46 (1H, s, 5-OH), 10.77 (1H, br.s, 7-OH), 9.74 (1H, br.s, 3-OH), 9.12 (1H, br.s, 4 -OH), 7.75 (1H, d, J = 1.8, H-2 ), 7.69 (1H, dd, J = 8.7, 1.8, H-6 ), 6.94 (1H, d, J = 8.7, H-5 ), 6.48 (1H, d, J = 1.7, H-8), 6.19 (1H, d, J = 1.7, H-6), 3.84 (3H, s, 3 -OCH3) [10]. Quercetin (6), 19.7 mg, yellow amorphous powder, C15H10O7, mp 297–299 C. ESI-MS, m/z: 301.5 [M – H] – [11]. Kaempferol (7), 7.3 mg, yellow amorphous powder, C15H10O6, mp 267–269 C. ESI-MS, m/z: 285.5 [M – H] –. 1Í NMR spectrum (400 MHz, DMSO-d6, , ppm, J/Hz): 12.48 (1H, s, 5-OH), 10.78 (1H, s, 7-OH), 10.10 (1H, s, 4 -OH), 9.37

Bioactive Constituents of Ziziphora clinopodioides

Ziziphora clinopodioides Lam. (Labiatae), a medicinal and edible plant, is mainly distributed in Xinjiang of China, Iran, Turkey, Mongolia, and Central Asia. It is commonly used in traditional Uyghur medicine for the treatment of fever, edema, neurasthenic, insomnia, tracheitis, lung abscess, hemorrhoids, hypertension, angina pectoris, coronary artery disease, and other cardiovascular diseases [1–4]. Phytochemical investigations on the genus Ziziphora have mainly focused on essential oil components as well as a few flavonoids, caffeoyl derivatives, fatty acids, phenolic acids, triterpenoids, and sterols [4]. In our previous studies, Z. clinopodioides Lam. was shown to acquire notable antihypertensive, antidiabetic, and antioxidant effects. In order to explore the antihypertensive constituents of this plant, the commonly adopted in vitro model of rat thoracic aortic rings [4] were applied for the bioassay-guided fractionation of active components from Z. clinopodioides Lam., with 11 compounds isolated from the 70% EtOH extract. Based on spectroscopic analyses, their structures were identified as diosmetin (1), apigenin (2), luteolin (3), caffeic acid (4), 5,7,2 -trihydroxyflavone 2 -O-D-glucopyranoside (5), methyl rosmarinate (6), betulinic acid (7), dibutyl phthalate (8), oleanolic acid (9), acacetin (10), and 5,6,4 -trihydroxy-7,8,3 -trimethoxyflavone (thymonin, 11), respectively, among which compounds 7 and 8 were obtained from the genus Ziziphora for the first time. Apigenin (2), luteolin (3), methyl rosmarinate (6), and oleanolic acid (9) were identified as potential vasorelaxant principles. Methyl rosmarinate (6), caffeic acid (4), and luteolin (3) possessed significant antioxidant capacities. Ziziphora clinopodioides along with the active principle oleanolic acid (9) were also reported to possess exciting antidiabetic properties herein for the first time.

CHEMICAL COMPOSITION OF Croton laevigatus

The tree Croton laevigatus Vahl. (Euphorbiaceae) is distributed mainly in China in the provinces of Yunnan, Guangdong, and Hainan. Roots and leaves of the plant are usually used by the Dai people as a traditional drug for treating fractures, malaria, stomach ache, and wounds received from falling [1]. Phytochemical studies of C. laevigatus have not yet been reported with the exception of diterpenoids with interesting skeletons [2, 3]. Air-dried ground leaves (20 kg) were extracted with MeOH (3 80 L) in order to obtain the crude extract (2120 g) after evaporation in vacuo. The extract was suspended in H2O (8.0 L) and fractionated successively with petroleum ether, CHCl3, and n-BuOH (3 8.0 L). The petroleum-ether extract (673 g) was chromatographed over a column of silica gel with elution by petroleum ether:EtOAc mixtures (1:0, 9:1, 4:1, 3:2, 0:1) to afford compounds 1 (3 g) and 5 (10 g). The BuOH fraction was chromatographed over a column of silica gel with gradient elution by CHCl3:CH3OH mixtures (50:1 0:1) to afford 2 (50 mg), 3 (5 mg), 4 (20 mg), and 6 (20 mg). PMR and 13C NMR spectra were used to identify 1–4. Compounds 5 and 6 were identified by direct comparison with authentic samples as -sitosterol and daucosterol, respectively. 3 -(4 -Hydroxy-3 ,5 -dimethoxyphenyl)propylbenzoate (1), brown oil, C18H20O5. EI-MS (m/z, %): 316 (90) [M]+, 194 (100), 179 (23), 167 (52), 163 (53), 105 (75), 77 (92). The PMR and 13C NMR spectra were published [4]. Dihydrodehydrodiconiferyl alcohol -D-glucoside (2), yellow semi-solid compound, C26H34O11. ESI-MS (m/z, +-ions): 545.2 [M + Na]+, 383.1 [M + Na – glucose]+, 365.1 [M + Na – glucose – H2O] +. The PMR and 13C NMR spectra were published [5]. Myrsinionoside C (3), yellow semi-solid compound, C19H36O7. ESI-MS (m/z, +-ions): 400.2 [M + Na] +. The PMR and 13C NMR spectra were published [6]. Alangionoside J (4), yellow semi-solid compound, C19H36O7. ESI-MS (m/z, +-ions): 400.2 [M + Na] +. The PMR and 13C NMR spectra were published [7]. All compounds were isolated from C. laevigatus for the first time whereas 2–4 were isolated for the first time from a plant of the genus Croton.