[Rapid determination of active components in Ginkgo biloba leaves by near infrared spectroscopy combined with genetic algorithm joint extreme learning machine].

Near-infrared spectroscopy(NIRS) combined with band screening method and modeling algorithm can be used to achieve the rapid and non-destructive detection of the traditional Chinese medicine(TCM) production process. This paper focused on the ginkgo leaf macroporous resin purification process, which is the key technology of Yinshen Tongluo Capsules, in order to achieve the rapid determination of quercetin, kaempferol and isorhamnetin in effluent. The abnormal spectrum was eliminated by Mahalanobis distance algorithm, and the data set was divided by the sample set partitioning method based on joint X-Y distances(SPXY). The key information bands were selected by synergy interval partial least squares(siPLS); based on that, competitive adaptive reweighted sampling(CARS), successive projections algorithm(SPA) and Monte Carlo uninformative variable(MC-UVE) were used to select wavelengths to obtain less but more critical variable data. With selected key variables as input, the quantitative analysis model was established by genetic algorithm joint extreme learning machine(GA-ELM) algorithm. The performance of the model was compared with that of partial least squares regression(PLSR). The results showed that the combination with siPLS-CARS-GA-ELM could achieve the optimal model performance with the minimum number of variables. The calibration set correlation coefficient R_c and the validation set correlation coefficient R_p of quercetin, kaempferol and isorhamnetin were all above 0.98. The root mean square error of calibration(RMSEC), the root mean square error of prediction(RMSEP) and the relative standard errors of prediction(RSEP) were 0.030 0, 0.029 2 and 8.88%, 0.041 4, 0.034 8 and 8.46%, 0.029 3, 0.027 1 and 10.10%, respectively. Compared with the PLSR me-thod, the performance of the GA-ELM model was greatly improved, which proved that NIRS combined with GA-ELM method has a great potential for rapid determination of effective components of TCM.

[Determination of 10 mycotoxin contaminants in Panax notoginseng by ultra performance liquid chromatography-tandem mass spectrometry].

To ensure the quality and safety of Panax notoginseng, a method for the simultaneous determination of 10 mycotoxins in Panax notoginseng was developed using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The sample was extracted with acetonitrile and purified by HLB multifunction cleanup column. The separation was performed on a Phenomenex Kinetex XB-C18 column by gradient elution using methanol and 5 mmol·L(-1) ammonium acetate as mobile phase. The targeted compounds were detected in MRM mode by mass spectrometry with electrospray ionization (ESI) source operated in both positive and negative ionization modes. The linear relationships of the 10 mycotoxins were good in their respective linear ranges. The correlation coefficients (r) ranged from 0.9981 to 1.0000. The LOQs of the 10 mycotoxins were between 0.15 and 8.6 µg·kg(-1). The average recoveries ranged from 73.8% to 107.0% with relative standard deviations (RSDs) of 0.10%-10.9%. The results demonstrated that the proposed method was sensitive and accurate, and suitable for the mycotoxins quantification in Panax notoginseng.

[Determination of four effective components from total flavonoids of Scutellaria barbata by high performance liquid chromatography].

OBJECTIVE: To establish a HPLC method for simultaneous determination of 4 effective components from total flavonoids of Scutellaria barbata (FSB). METHODS: The HPLC method was developed on an Agilent Zorbax C18 column (4.6 mm × 250 mm, 5 µm). The mobile phase was composed of 1% HAc and CH3OH:CH3CN (80:20) with a linear gradient elution. The flow rate was 1.0 ml/min, and UV detection wave length was set at 280 nm. The column temperature was maintained at 30°C. RESULT: The linear range of 4 effective components (scutellarin, isoscutellarein-8-O-glucuronide, isoscutellarein and luteolin) was 0.14-11.20 µg, 0.03-2.40 µg, 0.007-0.560 µg and 0.027-2.160 µg, respectively. The average recovery for 4 effective components was (101.9 ± 1.4)%, (103.5 ± 0.6)%, (98.1 ± 2.9)% and (100.5 ± 2.3)%, respectively. The contents of 4 flavonoids were determined, with scutellarin 7.3%-14.3%, isoscutellarein-8-O-glucuronide 2.4%-9.3%, isoscutellarein 0.3%-0.5%, and luteolin 0.2%-0.6%, respectively. CONCLUSION: The method can be used effectively to evaluate the quality of FSB.

[Comprehensive quality control method for total flavonoid of Fructus Aurantii].

OBJECTIVE: To establish a comprehensive quality control method for total flavonoid of Fructus Aurantii. METHODS: RP-HPLC and spectrophotometry were applied for the quantitative and fingerprint analysis of total flavonoid of Fructus Aurantii. The contents of naringin and neohesperidin were determined on an Agilent SB-C18column (4.6 mm × 250 mm, 5 µm). The mobile phase was composed of 0.02 % H3PO4 and CH3CN (80:20). The flow rate was 1 ml/min with DAD detected at 280 nm. The column temperature was maintained at 35°C. The fingerprints were developed on an Agilent SB-C18 column (4.6 mm × 250 mm, 5 µm). The mobile phase was composed of 0.5 % HAc and CH3OH with a linear gradient elution. The ratio of 0.5 % HAc and CH3OH was: 0 min, 80:20; 10 min, 60:40; 35 min, 30:70; 50 min, 0:100. The flow rate was 1 ml/min with DAD detected at 320 nm. The column temperature was maintained at 30 degree. Meanwhile, the contents of total flavonoid were determined at 283 nm. RESULT: The contents range of naringin, neohesperidin and total flavonoid were 38.3 %- 47.2%, 21.0 %- 28.5% and 79.9%-88.6 %, respectively. The fingerprints of the effective fractions showed 12 common peaks and the fingerprint similarity was all above 98.0 % compared with the standard chromatogram. CONCLUSION: The method reported in this paper can be used effectively for the quality control of total flavonoid of Fructus Aurantii.

Development of the fingerprint for the quality of Radix Linderae through ultra-pressure liquid chromatography-photodiode array detection/electrospray ionization mass spectrometry.

An ultra-pressure LC (UPLC) method has been developed and validated for the quality evaluation of a traditional Chinese medicine (Radix Linderae) by chemical fingerprint analysis with chromatograms collected at two wavelengths (260 and 320 nm). Eleven characteristic peaks in the fingerprints were identified by comparing their retention times, UV spectra and ESI-MS/MS data with those of the reference substances or the data in the literatures. Both correlation coefficient of similarities in chromatograms and relative peak areas of common peaks were calculated for quality expression of the Radix Linderae samples collected from different areas in China. The results showed high variation of relative peak area and correlation coefficients among the samples collected from various habitats, which indicated that the quality consistency of Radix Linderae is still a problem worthy of serious concern.

[Effect of griffithin on anticancer activity and apoptosis of cancer cells in vitro].

To study the anticancer activity of griffithin from Streptocaulon griffithii Hook. f. and its effect on apoptosis of cancer cells in vitro, the inhibitory effect of griffithin on cell proliferation was studied by MTT assay, the cell apoptosis was observed by AO/EB double decoration assay and flow cytometry. Griffithin exhibited high anticancer activity on four human cancer cell lines, with IC50 ranged from 0.17 - 0.43 microg x mL(-1). Griffithin also induced apoptosis of PC-3 cells. Griffithin had anticancer activity and induced apoptosis of cancer cells.

[Influence of H2O2 on degradation of residual pesticides and constituents in Radix Sophorae Flavescentis].

OBJECTIVE: To investigate the characteristic and influential factors of the degradation of residual pesticides and alkaloids in Radix Sophorae Flavescentis by H2O2. METHOD: The spiked samples were treated in H2O2 in different reaction time, concentration and pH value. The pesticide residuals were determined by GC-MS, and the contents of alkaloids were determined by HPLC. RESULT: H2O2 had highly activity in degrading organophosphorus and pyrethroid, but had less activity to organochlorines. The degradation processes of organophosphorus and pyrethroid followed first-order kinetics equations, and were influenced by the pH value, the concentration of H2O2 and reaction time. The contents of alkaloids in Radix Sophorae Flavescentis changed not obviously after treatment with 3 mL x L(-1) H2O2 less than 6 hours under neutral condition. CONCLUSION: H2O2 is a useful reagent for the degradation of organophosphorus and pyrethroid in crude drug.

[Effects of water extractives of a qi and blood regulating prescription on early atherosclerosis of apolipoprotein E-deficient mice].

OBJECTIVE: To investigate whether the water extractives of regulating qi and blood prescription (WQBP) had effects on early atherosclerosis of apolipoprotein E-deficient mice (ApoE-mice) at the age of 19 weeks or not, and to explore the possible mechanisms. METHODS: Forty ApoE-mice, six weeks of age, were given high-fat diet and randomly divided into four groups: high-dose WQBP-treated group (360 mg/kg), low-dose WQBP-treated group (72 mg/kg), simvastatin-treated group (25 mg/kg) and untreated group, with ten mice in each group. Meanwhile, ten C57BL/6 mice of same genetic background were allocated to normal control group. Mice in the high- and low-dose WQBP-treated groups and simvastatin-treated group were administered with corresponding drugs from the 15 to 19 weeks. Mice in the untreated and normal control groups were administered with isovolumic water. Sacrificed at 19 weeks, the level of blood-lipid, the plaque construction, plaque integral, and the contents of plaque macrophages and vessel smooth muscle cells of the mice were analyzed by immunohistochemical method and a computer picture processing system. RESULTS: Compared to the untreated group, high-dose WQBP group could obviously decrease the level of low-density lipoprotein cholesterol (LDL-C). Simvastatin group could decrease the levels of LDL-C and total cholesterol (TC) (P<0.01). In high-dose WQBP-treated group and simvastatin-treated group, the thickness of fiber cap and the quantities of vessel smooth muscle cells increased (P<0.05), the quantities of plaque macrophages and the ratio of lipid and plaque reduced (P<0.01). CONCLUSION: WQBP and simvastatin can interfere in early atherosclerosis of ApoE-mice, attenuate and stabilize plaque in some extent. The mechanisms may include adjusting blood lipid, decreasing macrophage number and increasing the quantities of vessel smooth muscle cells.

Stereoselective urinary excretion of S-(-)- and R-(+)-propranolol glucuronide following oral administration of RS-propranolol in Chinese Han subjects.

AIM: To study the stereoselectivity of phase II glucuronidation metabolism of side-chain propranolol in Chinese Han population. METHODS: Sixteen adult Chinese Han volunteers with an average age of 20 years were given a single oral dose of 20 mg racemic propranolol. Human urine at indicated time after administration was collected and S-(-)-propranolol glucuronide and R-(+)-propranolol glucuronide were determined simultaneously by using RP-HPLC. RESULTS: The mean values of k were 0.19+/-0.04 h(-1) and 0.28+/-0.06 h(-1), of t(1/2) 3.56+/-0.73 h and 2.45+/-0.50 h, of T(max) 2.21+/-0.45 and 1.75+/-0.33 h, and of Xu(0-24) 5.65+/-0.98 and 2.95+/-0.62 micromoL for S-(-)- and R-(+)-propranolol glucuronide, respectively. The cumulative excretion percentages in urine of doses were 14.7+/-2.46% and 7.68+/-1.60% for S-(-)- and R-(+)-propranolol glucuronide, respectively. The results showed the elimination rate constant k of S-(-)-propranolol glucuronide was less than that of R-(+)-propranolol glucuronide; and the elimination half-life (t(1/2)), T(max) and the cumulative excretion amount(Xu(0-24)) of R-(+)-propranolol glucuronide were significantly less than that of S-(-)-propranolol glucuronide. CONCLUSION: The propranolol glucuronidation of the side-chain undergoes stereoselective excretion in Chinese Han population after an oral administration of racemic propranolol.

[Study on quality control of effective fraction in qixue bingzhi decoction].

OBJECTIVE: To develop a method for quality control of effective fraction in Qi-Xue-Bing-Zhi decoction, a traditional Chinese medicine (TCM). METHOD: PF samples, effective fraction from Qi-Xue-Bing-Zhi decoction, were used as example, and a HPLC assay for chemical fingerprint and quantitative analysis was established. RESULT: The contents range of Paeoniflorin (PE), Naringin (NG) and Neohesperidin (NH) in effective fractions were changed from 12.5%-16.0%, 8.4%-12.4%, 12.8%-15.3%, and their average contents were (14.7 +/- 1.1)%, (10.6 +/- 1.2)%, (14.2 +/- 0.8)% (n = 10), respectively. The fingerprints of PF samples showed 25 common peaks, and the fingerprint similarity for PF samples were all above 99.00% by comparing with the standard chromatogram. CONCLUSION: The method reported could be used effectively for the quality control of effective fraction from TCM.

[Quantitative analysis of the nucleosides in Cordyceps sinensis with capillary zone electrophoresis].

OBJECTIVE: To establish a quantitative analysis method for analyzing the nucleosides in Cordyceps sinensis with capillary electrophoresis, and compare the difference between natural and the cultured C. mycelia. METHOD: Capillary zone electrophoresis method was employed to quantitate the adenosine, uridine, guanosine and inosine in C. sinensis, with 0.25 mg x L(-1) boric acid-sodium hydroxide buffer, pH 9.5. The working voltage was 20 kV, the temperature was 25 degrees C, and the detection wavelength was 260 nm. RESULT: With the capillary zone electrophoresis method, the average recovery of the above 4 nucleosides was 98.9%, 95.1%, 97.8% and 98.8% respectively, with the RSD 0.4%, 1.7%, 1.3% and 5.0%. There was no adenosine in natural C. sinensis and no inosine in the cultured C. mycelia detected. CONCLUSION: This method can be used to determine the adenosine, uridine, guanosine and inosine in C. sinensis. The nucleosides in C. sinensis produced from Qinghai province and cultured C. mycelia are obviously different.

[Effects of microsome enzyme induced by phenobarbarbital on the stereoselectivity of recemic propranolol glucuronidation metabolism].

OBJECTIVE: To study the stereoselectivity of R-(+) and S-(-)-propranolol glucuronidation and metabolic interaction between R(+)- and S-(-)-propranolol. METHODS: A RP-HPLC analytical method was developed for determination of R-(+)-and S-(-)-propranolol glucuronide (PG) incubated with rat hepatic microsome induced with phenobarbital (PB). The method was applied to investigate the stereoselectivity metabolism of racemic propranolol glucuronidation in vitro. RESULT: In control and PB group, the concentration of R-(+)-PG produced at different substrates was higher than that of S-(-)-PG. Compared with the control, the V(max) and Cl(int) for R(+)-and S-(-)-propranolol increased significantly the K(m) for R(+)-propranolol was elevated, while that for S-(-) propranolol was decreased. CONCLUSION: There is a stereoselectivity in glucuronidation of propranolol in rat hepatic microsome induced with PB and R-(+)-propranolol is preferred. Metabolic interaction between R-(+)-and S-(-)-propranolol exists with a concentration-dependent mode.

[The contents of paeoniflorin in different combination jingzhixuefuzhuyu].

OBJECTIVE: To investigate the contents of paeoniflorin in different combination Jingzhixuefuzhuyu. METHOD: Using RP-HPLC to determine the contents of paeoniflorin in different combination Jingzhixuefuzhuyu extracts, an ODS column was used with a mobile phase of MeOH-H2O-HAC(25:75:0.15) and DAD detector at the wavelength of 230 nm. RESULT AND CONCLUSION: Different combinations of Jingzhixuefu zhuyu had great influence to the contents of paeoniflorin.