ABSTRACT
To study the quality control of three traditional Chinese medicines derived from Gleditsia sinensis [Gleditsiae Sinensis Fructus(GSF), Gleditsiae Fructus Abnormalis(GFA), and Gleditsiae Spina(GS)], this paper established a multiple reaction monitoring(MRM) approach based on ultra-high performance liquid chromatography-triple quadrupole-linear ion-trap mass spectrometry(UHPLC-Q-Trap-MS). Using an ACQUITY UPLC BEH C_(18) column(2.1 mm × 100 mm, 1.7 μm), gradient elution was performed at 40 ℃ with water containing 0.1% formic acid-acetonitrile as the mobile phase running at 0.3 mL·min~(-1), and the separation and content determination of ten chemical constituents(e.g., saikachinoside A, locustoside A, orientin, taxifolin, vitexin, isoquercitrin, luteolin, quercitrin, quercetin, and apigenin) in GSF, GFA, and GS were enabled within 31 min. The established method could quickly and efficiently determine the content of ten chemical constituents in GSF, GFA, and GS. All constituents showed good linearity(r>0.995), and the average recovery rate was 94.09%-110.9%. The results showed that, the content of two alkaloids in GSF(2.03-834.75 μg·g~(-1)) was higher than that in GFA(0.03-10.41 μg·g~(-1)) and GS(0.04-13.66 μg·g~(-1)), while the content of eight flavonoids in GS(0.54-2.38 mg·g~(-1)) was higher than that in GSF(0.08-0.29 mg·g~(-1)) and GFA(0.15-0.32 mg·g~(-1)). These results provide references for the quality control of G. sinensis-derived TCMs.
Subject(s)
Flavonoids/analysis , Alkaloids , Chromatography, High Pressure Liquid/methods , Mass Spectrometry , Drugs, Chinese HerbalABSTRACT
In view of the current inadequate standards for Gleditsiae Spina in the Chinese Pharmacopoeia, this study put forward some new items of the quality standards of Gleditsiae Spina. Thin-layer chromatography(TLC) was performed for identification with the reference substance of taxifolin and the reference material of Gleditsiae Spina as the control. According to the general principles of the Chinese Pharmacopoeia(2020 edition, Vol. 4), the moisture, total ash content, and alcohol-soluble extract of medicinal materials and decoction pieces of Gleditsiae Spina were determined. The content determination method for medicinal materials and decoction pieces of Gleditsiae Spina was established using high-performance liquid chromatography(HPLC), with taxifolin as the quality control index. Based on the determination results of 30 batches of samples of Gleditsiae Spina from different habitats, the draft quality standards of Gleditsiae Spina were developed, which provided suggestions for the revision of the quality standards of Gleditsiae Spina in the Chinese Pharmacopoeia.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Quality Control , Reference StandardsABSTRACT
The chemical constituents from ethyl acetate extract of Gleditsiae spina were isolated and purified by various chromatographic methods such as MCI gel CHP-20, ODS, Sephadex LH-20, silica gel and semi-preparative HPLC. Seven lignans were isolated and identified by spectroscopic data analyses as (7R,8S,7'E,7''S,8''R)-buddlenol P (1), (+)-syringaresinol (2), (+)-isolariciresinol (3), (7S,8R)-cedrusin (4), (7S,8R)-4,9,9'-trihydroxy-3,3'-dimethoxy-7,8-dihydrobenzofuran-1'-propylneolignan (5), 3',4-O-dimethylcedrusin (6), balanophonin (7). Among them, compound 1 is a new lignan, compounds 2-7 are isolated from the Gleditsia L. for the first time. MTT method was used to investigate the effect of compounds 2-7 on LPS-induced injury of NRK-52e cells. As a result, compounds 2, 3 and 7 exhibit protective effects against LPS-induced damage to NRK-52e cells.
ABSTRACT
Objective To investigate the chemical constituents from the ethyl acetate extract of Gleditsiae Spina. Methods The chemical constituents were isolated and identified by chromatography on silica gel, Toyopearl HW-40C, Sephadex LH-20, MCI Gel CHP-20, ODS, and RP-HPLC. Their structures were elucidated on the basis of physicochemical properties and spectral analyses. Results Eighteen compounds were isolated from the ethyl acetate extract of Gleditsiae Spina, and identified as scopoletin (1), (-)-(7R,8S)-erythro-guaiacylglycerol (2), 5-methoxy-guaiacylglycerol (3), (-)-(7R,8R)-threo-guaiacylglycerol (4), 3,4’- dihydroxypropiophenone (5), dihydroferulic acid (6), p-hydroxybenzoic acid (7), indole-3-carbaldehyde (8), protocatechuic acid methyl (9), protocatechuic aldehyde (10), syringic acid (11), 2-guaiacylpropane-1,3-diol (12), (2R*,3R*,4S*)-2,3-diguaiacyl-4- hydroxyl tetrahydrofuran (13), 3-(2-oxopropyl)-3-hydroxy-indolin-2-one (14), juglanin D (15), C-veratroylglycol (16), 3-(4-hydroxyl-3-methoxyphenyl)-propan-1,2-diol (17), and (-)-syringaresinol (18). Conclusion Eighteen compounds are isolated from this plant for the first time.
ABSTRACT
Objective: An HPLC method was developed to evaluate the quality of Gleditsiae Spina through three components determination and fingerprint analysis. Methods: The assay was performed on Waters symmetry C18 (250 mm × 4.6 mm, 5 μm) column. The mobile phase consisted of acetonitrile and 0.1% methanoic acid in water with gradient elution by PDA detection at 338 nm. Results: Thirteen batches of Gleditsiae Spina were analyzed with the established method. In the fingerprint, 14 common peaks were marked and three of them were determined. The three compounds showed good linearity (r ≥ 0.999 2) in the range of 0.091 3-5.840 0, 0.176 3-11.280 0, and 0.014 0-0.895 0 mg/mL, respectively. The average recoveries were 98.97%-99.66% with RSD < 2.5%. Conclusion: This method has good repeatability and stability and provides a new method for the quality control of Gleditsiae Spina.
ABSTRACT
Objective: To investigate the chemical constituents from Gleditsiae Spina (the thorns of Gleditsia sinensis). Methods: The compounds were isolated and purified by silica gel, Sephadex LH-20 column chromatographic techniques, and their chemical structures were confirmed on the basis of spectroscopic analysis by the physicochemical properties. Cytotoxic activity using the MTT colorimetry method was performed to measure the inhibitory effect of the compounds on cell proliferation of HepG2, A-549, and EC109. Results: Three coumarins were obtained from the ethyl acetate soluble fraction of the 90% ethanolic extract and their structures were identified as scoparone (1), isoscopoletin (2), and 6-amino-7-methoxycoumarin (3). Conclusion: Compound 3 is a novel coumarin named gledisinmarin A. Compounds 1 and 2 are isolated from this plant for the first time. Compound 2 displays the stronger cytotoxicity against A549 cell with an IC50 value of 34.47 μg/mL, while cisplatin with an IC50 value of 11.50 μg/mL as a positive control.
ABSTRACT
Objective: To investigate the flavonoids from Gleditsiae Spina (thorns of Gleditsia sinensis) and their antitumor activities. Methods: The chemical constituents in the EtOAc fraction from Gleditsiae Spina were isolated and purified by the chromatography on silica gel, Sephadex LH-20 columns, and semi-preparative HPLC. Their structures were identified by various spectroscopic analyses, and the cytotoxicity of compounds 7-16 were evaluated in vitro against MCF-7 cell lines by SRB method. Results: Sixteen compounds were isolated from the extracts of Gleditsiae Spina and identified as tricin (1), liquiritigenin (2), 7,4'-dihydroxy-5,3'-dimethoxyflavanonol (3), garbanzol (4), 7,3',5'-trihydroxyflavanone (5), 7,4'-dihydroxyflavonol (6), dihydrokaempferol (7), butein (8), (2S)-5,7,3',5'- tetrahydroxyflavanone (9), 7,3',5'-trihydroxy-5-methoxyflavanonol (10), quercetin (11), fustin (12), fisetin (13), leucorobinetinidin (14), thevetiaflavon (15), and isovitexin (16). Compound 8 showed the inhibitory effect against MCF-7 cells with IC50 value of 28.53 μmol/L. Conclusion: Compounds 1-6, 8-10, 14 and 15 are isolated from the plants of Gleditsia L. for the first time, compound 8 shows the significant cytotoxic activity against MCF-7 cells.
ABSTRACT
Objective: To study the flavonoids constituents from the thorns of Gleditsia sinensis and their cytotoxicity against tumor cells. Methods: The compounds were isolated by silica gel, Sephadex LH-20 column chromatography, and HPLC techniques. Their structures were elucidated on the basis of spectroscopic analyses. Results: Twelve flavonoids were obtained and identified as (2R,3R)-5,3',4'-trimethoxyl-7-hydroxyl-flavanonol (1), 5,7,3',4'-tetrahydroxyl-flavanonol (2), 5-methoxyl-3',4',7-trihydroxyl-flavanonol (3), dihydrokaempferol (4), epicatechin (5), 5,7,3',5'-tetrahydroxyl-flavanonol (6), fustin (7), (2R,3R)-7,3',5'-trihydroxyl-flavanonol (8), (2R,3R)-5,7,3'-trihydroxyl-4'-methoxyl-flavanonol (9), quercetin (10), 5,7,4'-trihydroxylflavone-8-C-glucopyranose (11), and 2,7-dimethyl-xanthone (12), respectively. Conclusion: Compound 1 is a new component named G. spina flavanonol A, and compounds 3,8,9, and 12 are isolated from the thorns of G. sinensis for the first time. The results of cytotoxicity test show that the dihydroflavonol compound 7 displays the stronger cytotoxicity against HepG2, A549, and EC109 cell strains, while compounds 1 and 3 have the effects on HepG2 and EC109, and compound 2 has the effect on EC109 cancer cells, respectively.
ABSTRACT
Objective: To investigate the chemical constituents from Gleditsiae Spina (the thorns of Gleditsia sinensis) and their antitumor activitives. Methods: The chemical constituents were isolated and purified by the chromatography on repeated silica gel, Sephadex LH-20 column, semi-preparative HPLC, etc. Their structures were elucidated by NMR and MS spectroscopic data analyses; The cytotoxicity of compounds 1-4 and 6 was evaluated against human liver cancer SK-HEP-1 cells by MTT assay. Results: Twelve compounds were isolated from Gleditsiae Spina, and identified as 2-aminoimidazole (1), 2,3-dihydro-5-(2-formylvinyl)-7-hydroxy-2-(4-hydroxy-3-methoxyphenyl)-3-benzofuranmethanol (2), E-cinnamic acid (3), 3-O-trans-feruloylquinic acid (4), trans-caffeic acid (5), 4-hydroxy-3-methoxybenzamide (6), threo-guaiacylglycerol-β-coniferyl aldehyde ether (7), 5,7-dihydroxy-chromone (8), vanillic acid (9), protocatechuic acid (10), 3-O-caffeoylquinic acid methyl ester (11), and 3-O-caffeoylquinic acid ethyl ester (12); Compound 1 exhibited the potent cytotoxicity against SK-HEP-1 cells with IC50 value of 34.47 μg/mL. Conclusion: All the compounds except compound 5 are isolated from the plants of Gleditsia L. for the first time; Compound 1 shows significant cytotoxic activity against SK-HEP-1 cells.
ABSTRACT
Objective: To explore the prevention and treatment of total flavonoids from Gleditsiae Spina (TFGS) on lung cancer and its mechanisms. Methods: Mouse Lewis lung cancer (LLC) and embryonic lung fibroblast (L929) cells were treated with different doses of TFGS for 48 h, cell proliferation and adhesion were examined by MTT assay, and gap junctional intercellual communication (GJIC) was measured through scrape loading and dye transfer. The mice were randomly divided into model, quercetin (100 mg/kg, positive control), high-and low-dose (100 and 30 mg/kg) TFGS groups. The mice were ip injected with urethane twice weekly for five weeks to induce lung carcinogenesis and treated once daily for 10 weeks following the first urethane injection. The prevention of TFGS on chemocarcinogenesis was evaluated and the expression of gap junctional protein connexin 43 (Cx43) in lung tissue with tumors was compared by immunohistochemistry. The LLC cells were injected into the lateral axilla and tail vein respectively to establish the LLC sc allograft and experimental lung metastasis. The tumor-inocubating mice were randomly divided into model, doxorubicin (5 mg/kg, positive control), high-and low-dose (same as above) TFGS groups. The mice received the treatment for three weeks following tumor inocubation, and the effects of TFGS on the tumor size, metastasis, and life span were evaluated. Results: TFGS inhibited LLC cell proliferation in a dose dependent manner but had no effect on L929 cell proliferation in vitro. TFGS with a little effect on cell proliferation decreased cell adhesion and promoted GJIC in a dose dependent manner in LLC cells but did not affect the L929 cell adhesion. TFGS was able to prevent carcinogenesis induced by urethane and enhance Cx43 staining in lung region with tumor in immunohistochemistry. Compared with untreated model mice, GJIC reduced the tumor size and metastasis and prolongated life span in a dose dependent manner. Conclusion: TFGS could promote GJIC to prevent and treat tumor and might be a potential antitumor agent.