OBJECTIVETo establish an UPLC method for simultaneous determination of purpuroxanthine, purpurin, 1,3,6-trihydroxy-2-methylanthraquinone, rubimaillin in carbonized Rubiae Radix et Rhizoma.

METHODThe components were separated on acquity BEHC18 (2.1 mm x 50 mm, 1.7 microm) using methanol and 0.3% formic acid solution as the mobile phase; The flow rate was 0.2 mL x min(-1) and the volume of injection was 2 microL; the column temperature was maintained at 30 degrees C and the detective wavelength was set at 276 nm.

RESULTThere were good liner relationships between the peak area and concentration at ranges of 0.68-34.44 mg x L(-1) (r = 0.9999), 0.66-33.2 mg x L(-1) (r = 0.9997), 0.68-34.08 mg x L(-1) (r = 0.9999), 1.07-53.52 mg x L(-1) (r = 0.9999) for purpuroxanthine, purpurin, 1,3,6-trihydroxy-2-methylanthraquinone, rubimaillin, respectively; the average recovery rates of purpuroxanthine, purpurin, 1,3,6-trihydroxy-2-methylanthraquinone, rubimaillin were 96.95%, 95.75%, 102.5%, 96.15%, respectively, with RSD less than 3%.

CONCLUSIONThe established method was rapid and simple with good accuracy and reproducibility for the determination of carbonized Rubiae Radix et Rhizoma, the method was suitable for the quality control of carbonized Rubiae Radix et Rhizoma.

Chromatography, High Pressure Liquid ; Quinones ; chemistry ; Rhizome ; chemistry ; Rubia ; chemistry

Fourteen compounds, including rubiprasin D (1), rubiprasin B (2), rubiprasin C (3), oleanolic acid (4), methyl-5-hydroxy-dinaphtho[1, 2-2'3']furan-7, 12-dione-6-carboxylate (5), rubioncolin C (6), mollugin (7), furomollugin (8), 3-amino-2-methoxycarbonyl-1, 4-naphthoquinone (9), 1-hydroxy-2-methyl-9, 10-anthraquinone (10), 2-hydroxy-6-methyl-9, 10-anthraquinone (11), 1, 4-dihydroxy-2-hydroxymethyl-9, 10-anthraquinone (12), 2-hydroxy-1-methoxy-9, 10-anthraquinone (13), and 1-hydroxy-2-methoxy-6-methyl-9, 10-anthraquinone(14), were isolated from the methanol extract of the roots and rhizomes of Rubia oncotricha using various column chromatographies. Their structures were mainly determined on basis of NMR and MS spectroscopic data analyses. Among them, 1 is a new oleanane triterpene, and compounds 2-5, 9 and 11-13 were obtained from this plant for the first time. Cytotoxic and nematicidal activities of all these compounds were evaluated, and the results showed that only 4, 6, 11 and 12 exhibited cytotoxicities against A549, SGC-7901 and HeLa cancer cell lines. The IC₅₀ of 6 were 19.42, 2.74, 8.07 μmol·L⁻¹, respectively.
Molecular Structure ; Naphthoquinones ; Plant Extracts ; Plant Roots ; Rhizome ; Rubia

Four kinds of ionic liquids were adopted to analyze the content of rubimaillin and alizarin in Rubia cordifolia roots with ultrasonic-assisted extraction coupled with HPLC. The chromatographic column, Purospher star RP-C18 (4.6 mm x 250 mm, 5 microm), was used. Methanol and 0.4% acetic acid-water as mobile phase with flow rate at 0.85 mL min(-1), gradient elution, detection wavelength at 250 nm, chromatographic column temperature was controlled at room temperature. The result showed that rubimaillin and alizarin had the highest extraction yield when the [ HMIM] PF6methanol solution concentration of 0.6 mol x L(-1) as extraction solvent and the conditions were solid-liquid ratio of 1:80 (g x mL(-1)). Under the optimal extraction conditions, the content of alizarin from 0.01 to 0.04 microg showed a good linearity (r = 0.9999), the average recovery was 97.12%, the content of rubimaillin from 0.41 to 1.35 microg showed a good linearity (r = 0.9999), the average recovery was 98.10%. This experiment adopted environmentally friendly reagent as extraction solvent, the extraction efficiency was improved, and the environmental pollution caused by organic solvent was avoided, the harm of human body aslo was reduced. This method was simple and reliable, its repeatability was also very good, which had an important significance in the study of traditional Chinese medicine active ingredient extraction methods.
Anthraquinones ; analysis ; Chromatography, Reverse-Phase ; methods ; Ionic Liquids ; chemistry ; Pyrans ; analysis ; Rubia ; chemistry ; Ultrasonics

OBJECTIVETo separate and identify cyclopeptides of tubers of Rubia schumanniana.

METHODThe 70% methanol extracts from tubers of Rubia schumanniana were separated and purified by silica gel, RP-18, Sephedax LH-20 and HPLC. Their structures were identified by spectral analysis.

RESULTNine cyclopeptides were separated and identified as RA- II (1), RA-V (2), RA-VIII (3), rubiyunnanin C (4), RA-X (5), RY-II (6), RA- I (7), RA-XIII (8) and RA-XIII-OMe (9), respectively.

CONCLUSIONAll of nine cyclopeptides were separated from R. schumanniana for the first time.

Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Peptides, Cyclic ; analysis ; isolation & purification ; Rubia ; chemistry

OBJECTIVETo search alpha-glucosidase inhibitors from Rubia cordifolia.

METHODThe alpha-glucosidase inhibitors were isolated by the column chromatographic techniques and the bioassay-guided method in vitro. A combination of MS and NMR spectroscopy was used to identify the chemical structures. The inhibitory kinetics of the inhibitors were also investigated.

RESULTThe chloroform extract showed high inhibitory activity, and three active compounds were isolated and identified as 1,3-dihydroxy-2-methylanthraquinone (1), 1-hydroxy-2-methylanthraquinone (2) and 1,2-dihydroxyanthraquinone (3). The IC5o values of compound 1-3 were all lower than that of acarbose. Compound 1 and 2 shown competitive type manner on alpha-glucosidase, whereas compound 3 shown noncompetitive type model.

CONCLUSIONCompounds 1-3 as strong inhibitors of alpha-glucosidase were reported for the first time.

Binding, Competitive ; Enzyme Inhibitors ; analysis ; metabolism ; pharmacology ; Glycoside Hydrolase Inhibitors ; Inhibitory Concentration 50 ; Kinetics ; Rubia ; chemistry ; alpha-Glucosidases ; metabolism

AIM: To determine the IPP origin of the naphthoquinones (NQs) in Rubia cordifolia, and to evaluate the effects of methyl jasmonate (MeJA) treatment, MEP, and MVA pathway inhibitor treatment on the accumulation of anthraquinones (AQs) and NQs in cell suspension cultures of R. cordifolia. METHODS: Cell suspension cultures of R. cordifolia were established. Specific inhibitors (lovastatin and clomazone) and MeJA were supplied to the media, respectively. Treated cells were sampled every three days. Content determination of purpurin (AQs) and mollugin (NQs) were carried out using RP-HPLC. The yield of the two compounds was compared with the DMSO-supplied group and the possible mechanism was discussed. RESULTS: Lovastatin treatment increased the yield of purpurin and mollugin significantly. Clomazone treatment resulted in a remarkable decrease of both compounds. In the MeJA-treated cells, the purpurin yield increased, meanwhile, the mollugin yield decreased compared with control. CONCLUSION The IPP origin of mollugin in R. cordifolia cell suspension cultures was likely from the MEP pathway. To explain the different effects of MeJA on AQs and NQs accumulation, studies on the regulation and expression of the genes, especially after prenylation of 1,4-dihydroxy-2-naphthoic acid should be conducted.
Acetates ; pharmacology ; Anthraquinones ; metabolism ; Cell Culture Techniques ; Cells, Cultured ; Cyclopentanes ; pharmacology ; Isoxazoles ; pharmacology ; Lovastatin ; pharmacology ; Oxazolidinones ; pharmacology ; Oxylipins ; pharmacology ; Pyrans ; metabolism ; Rubia ; drug effects ; metabolism

OBJECTIVETo observe the effects of Rubiae Radix et Rhizoma (RRR) and carbonized Rubiae Radix et Rhizoma (CRRR) on the acute blood stasis rat model, and reveal their differences in efficacy.

METHODThe acute blood stasis model was induced by subcutaneously injecting adrenaline hydrochloride and soaking in ice water. Yunnan Baiyao was used as the positive control drug, and administered for consecutively seven days. This model was adopted to observe the effect of high, middle and low dose RRR and CRRR groups on hemorheology, thrombin activity, and blood platelet system.

RESULTRRR could significantly reduce the wholeblood viscosity and plasma viscosity of blood stasis rats under different shear rates, and showed certain two-way regulating function in hemostasis. It also showed certain effect on ADP-induced platelet aggregation rate, but which was lower than CRRR. CRRR achieved the main hemostatic mechanism by stimulating intrinsic and extrinsic blood coagulation and fibrinogen, and could significantly enhance the platelet aggregation rate of rats in the acute blood stasis model (P <0. 01).

CONCLUSIONRRR had the effect of removing blood stasis and hemostasis, while CRRR mainly has the hemostatic effect. This further demonstrates the traditional processing theory of "promoting blood circulation with crude herbs and stopping bleeding with processed herbs".

6-Ketoprostaglandin F1 alpha ; blood ; Animals ; Blood Coagulation ; drug effects ; Carbon ; chemistry ; Chemistry, Pharmaceutical ; methods ; Disease Models, Animal ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Hemodynamics ; drug effects ; Male ; Medicine, Chinese Traditional ; methods ; Rats ; Rats, Sprague-Dawley ; Rubia ; chemistry ; Thromboxane B2 ; blood

OBJECTIVEConfirm the irritation of needle-like calcium oxalate crystals in raw Pinellia ternata.

METHODComparing the irritations of raw P. ternate containing calcium oxalate crystals, the raw P. ternate no containing calcium oxalate crystals, the pure needle-like calcium oxalate crystals isolated from raw P. ternata, the extracts of water and various solvents from raw P. ternate. by using the model of rabbits' eyes. Studying the quantity effect relationship of different concentration suspensions of needle-like calcium oxalate crystal isolated from raw P. ternate on rabbits' eyes. Observing the shape and appearance of calcium oxalate crystals in raw P. ternate and raw India Madder Root by the electro microscope and comparing their irritation degrees with the same contents of calcium oxalate crystals.

RESULTCalcium oxalate crystals in raw P. ternata showed very strong irritation property. Under the same content of calcium oxalate crystals, the irritation of raw P. ternata and pure needle-like calcium oxalate crystals isolated from raw P. ternate had no significant difference. The concentrations of needle-like calcium oxalate crystals were do relative to the degree of irritation on rabbits' eyes and they showed undoubted quantity-effect relationship.

CONCLUSIONCalcium oxalate crystal is the irritation component in raw P. ternata.

Animals ; Calcium Oxalate ; chemistry ; isolation & purification ; toxicity ; Conjunctiva ; drug effects ; Cornea ; drug effects ; Crystallization ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; isolation & purification ; toxicity ; Edema ; chemically induced ; Eye Diseases ; chemically induced ; Female ; Iris ; drug effects ; Male ; Pinellia ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rabbits ; Random Allocation ; Rubia ; chemistry