A study on the rapidly non-destructive detection method of Huoxiang Zhengqi oral liquid using near infrared spectroscopy / 药学学报

Based on near infrared spectroscopy and high performance liquid chromatography, this paper established the regression relationship between near infrared spectroscopy and index component content of Huoxiang Zhengqi oral liquid, so as to realize the rapid detection of index component content based on near infrared spectroscopy. Magnolol, honokiol and hesperidin were used as the quality indexes of Huoxiang Zhengqi oral liquid. After using the first derivative and normalization pretreatment method, characteristic variables were screened by CARS, and the correction model was finally established by partial least-squares regression (PLSR) method. The method accuracy was evaluated with the external validation, and the prediction results were tested for significance. The results indicated that when the near infrared spectrum was scanned through the bottle, the model's correlation coefficients of prediction (Rp) were higher than 0.99, the root mean square errors of the prediction models (RMSEP) were all less than 0.008 4, and the relative standard errors of prediction set (RSEP) were all less than 2.83%. There was no significant difference in the predicted results between these two kinds of model. The models established in the non-destructive way have good performance and high prediction accuracy. The rapid and nondestructive way has application value in the quality control of Huoxiang Zhengqi oral liquid.

Application of near infrared spectroscopy to predict contents of various lactones in chromatographic process of Ginkgo Folium / 中国中药杂志

This study established a method for rapid quantification of terpene lactone, bilobalide, ginkgolide C, ginkgolide A and ginkgolide B in the chromatographic process of Ginkgo Folium based on near infrared spectroscopy(NIRS). The effects of competitive adaptive reweighting sampling(CARS), random frog(RF), and synergy interval partial least squares(siPLS) on the performance of partial least squares regression(PLSR) model were compared to the reference values measured by HPLC. Among them, the correlation coefficients of prediction(Rp) of validation sets of terpene lactone, bilobalide, and ginkgolide C were all higher than 0.98, and the relative standard errors of prediction(RSEPs) were 5.87%, 6.90% and 6.63%, respectively. Aiming at ginkgolide A and ginkgolide B with relatively low content, the genetic algorithm joint extreme learning machine(GA-ELM) was used to establish the optimized quantitative analysis model. Compared with CARS-PLSR model, the CARS-GA-ELM models of ginkgolide A and ginkgolide B exhibited a reduction in RSEP from 15.65% to 8.52% and from 21.28% to 10.84%, respectively, which met the needs of quantitative ana-lysis. It has been proved that NIRS can be used for the rapid detection of various lactone components in the chromatographic process of Ginkgo Folium.

Rapid determination of active components in Ginkgo biloba leaves by near infrared spectroscopy combined with genetic algorithm joint extreme learning machine / 中国中药杂志

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.

Effects of shikonin on colon cancer xenografts in nude mice based on serum metabolomics / 药学学报

To investigate the antitumor activity of shikonin against human colorectal cancer, the IC50 value towards four different human colon cancer cells was detected by MTT assay. In addition, a SW620 xenograft model was established and both the tumor volume and tumor inhibitory rate were calculated to evaluate the antitumor activity of shikonin in vivo. To further explore the mechanism of shikonin, metabolomics combined with multivariate statistical analysis was performed to analyse the profile of metabolites in mouse serum. The results show that shikonin can significantly inhibit the proliferation of four different colon cancer cell lines and exerted a high antitumor activity in vivo. The tumor inhibitory rate at low dose and high dose were 38.35% and 42.16%, respectively. In addition, a total of 38 potential biomarkers related to the antitumor effects of shikonin were identified through metabolomics analysis, including tryptophan, proline and methionine. The study revealed that the mechanism was related to disordered amino acid metabolism in colon cancer, especially in tryptophan metabolism. Our study suggests that shikonin could exert an antitumor effect by regulating amino acid metabolism in colon cancer and provides a theoretical foundation for further exploration and the eventual clinical application of shikonin.

Simultaneous determination of five saponins in Yaobitong capsule by HPLC-CAD / 药学学报

A high performance liquid chromatography charged aerosol detector (HPLC-CAD) method was established for the simultaneous determination of five saponins (notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, Ginsenoside Rd) in Yaobitong capsule, providing a method for quality control. The sample was extracted with methanol and chromatographic separation was performed on a Waters Xbridge Phenol column (150 mm×4.6 mm, 3.5 μm) using acetonitrile-water as the mobile phase with gradient elution at a flow rate of 1.0 mL·min-1. The column temperature was 30 ℃ and the injection volume was 10 μL. The nebulizer temperature of CAD was 35 ℃ and the air pressure was 60.2 psi, the filtration was 3.6 s, and the collection frequency was 5 Hz. Notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ginsenoside Rd showed a good linear relationship in the range of 16.96-203.5 μg·mL-1 (R2 = 0.999 3), 54.46-653.5 μg·mL-1 (R2 = 0.999 3), 10.96-131.5 μg·mL-1 (R2 = 0.999 6), 51.50-618.0 μg·mL-1 (R2 = 0.999 0), 15.94-191.3 μg·mL-1 (R2 = 0.999 4), respectively. The average recoveries were 98.96%, 100.8%, 94.76%, 100.1%, 103.1%, and RSDs were 0.87%, 1.46%, 1.85%, 2.06%, 0.96% (n = 6), respectively. The proposed method is accurate, simple and reliable, and can be used for the determination of five saponins in Yaobitong capsule.

Fingerprint analysis and multi-component determination of Leonurus japonicus by high performance liquid chromatography / 药学学报

An HPLC fingerprint and multi-component determination method of Leonurus japonicus was established for comprehensive evaluation and quality control of Leonurus japonicus. The sample was incubated in 70% ethanol in a water bath for 2 h, and the extract was analyzed by HPLC using Kromasil C18 column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-0.1% formic acid with gradient elution. The flow rate was 1.0 mL·min-1. The temperature of column was 30 ℃. The detection wavelength was 280 nm. HPLC fingerprint of characteristic components of Leonurus japonicus was established. There were 12 common peaks among 25 batches of samples, and 5 of them were identified and determined. Syringic acid, leonurine hydrochloride, rutin, hyperoside or isoquercitrin showed a good linearity in the ranges of 0.426 1-85.22 ng (r = 0.999 9), 7.948-1 590 ng (r = 0.999 3), 10.20-2 040 ng (r = 1.000 0), 2.018-403.6 ng (r = 0.999 9), or 8.704-1 741 ng (r = 0.999 9), respectively. The average recoveries were 99.0%, 97.6%, 97.4%, 96.9% and 98.5% with RSD of 1.1%, 1.8%, 1.4%, 1.5% and 1.3%, respectively. The HPLC characteristic fingerprint of Leonurus japonicus was specific, and this method can simultaneously determine the content of 5 components.

The prediction of baicalin content in the extraction process of Scutellaria baicalensis by near-infrared spectroscopy combined with different variable selection methods / 药学学报

Near-infrared spectroscopy (NIRS) combined with chemometrics can achieve rapid detection in process analysis. After variable selection, the redundant information is effectively removed and the characteristic variables related to the response values are selected. Compared with global model, the complexity is significantly reduced and the prediction accuracy is also improved. In this study, near-infrared spectroscopy analysis combined with different variable selection methods was applied to achieve the rapid detection of baicalin in the extraction process of Scutellaria baicalensis. Data sets were divided based on sample set portioning based on joint x-y distance (SPXY) method. Competitive adaptive weighted resampling method (CARS), random frog (RF) and successive projections algorithm (SPA) were applied to variable selection. Partial least squares (PLS) models were constructed based on above three methods, and the prediction results were compared. After CARS, RF and SPA method, 92, 10 and 17 variables were screened out respectively. According to the performance of the models, CARS method is found to be more effective and suitable than RF and SPA. Furthermore, the characteristic variables selected by CARS method have a better correspondence with the chemical structure of baicalin. The root mean square error (RMSEC) of the calibration set and the root mean square error (RMSEP) of the prediction set are 0.528 2 and 0.720 2 respectively. Compared with the global PLS model, the correlation coefficient of the calibration set (Rc) is increased to 0.979 9 from 0.917 0, and the relative standard errors of prediction (RSEP) is reduced to 5.59% from 10.58%.

On-line detection of concentration process of Ganmaoling granules by near infrared spectroscopy combined with automatic control system / 药学学报

Our research was designed for on-line detection of multi-index in the concentration process of Ganmaoling granules by integration of near infrared spectroscopy and automatic control system. First, on-line detection system was set up in the concentration tank for Ganmaoling granules production. Spectra were scanned and values of chlorogenic acid, linarin, solid content and relative density were measured. Models of partial least squares regression were built and imported into near infrared workstation. By connecting the control system, real-time multi-index values were determined automatically in the concentration process. Results showed that correlation coefficients of chlorogenic acid, linarin, solid content and relative density models were 0.963, 0.989, 0.993 and 0.918, respectively. Relative standard errors of prediction were 3.71%, 4.28%, 4.17% and 0.24%, respectively, indicating a good performance and high accuracy of the models. Real-time data collection during the whole process was measured by the near infrared detecting system in the control system. In conclusion, the near infrared detection system is able to perform real-time automatic determination of multi-index in the concentration process of Ganmaoling granules with significant advantages.

Determination of 10 mycotoxin contaminants in Panax notoginseng by ultra performance liquid chromatography-tandem mass spectrometry / 药学学报

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.

Application of near infrared spectroscopy combined with particle swarm optimization based least square support vactor machine to rapid quantitative analysis of Corni Fructus / 药学学报

A novel method was developed for the rapid determination of multi-indicators in corni fructus by means of near infrared (NIR) spectroscopy. Particle swarm optimization (PSO) based least squares support vector machine was investigated to increase the levels of quality control. The calibration models of moisture, extractum, morroniside and loganin were established using the PSO-LS-SVM algorithm. The performance of PSO-LS-SVM models was compared with partial least squares regression (PLSR) and back propagation artificial neural network (BP-ANN). The calibration and validation results of PSO-LS-SVM were superior to both PLS and BP-ANN. For PSO-LS-SVM models, the correlation coefficients (r) of calibrations were all above 0.942. The optimal prediction results were also achieved by PSO-LS-SVM models with the RMSEP (root mean square error of prediction) and RSEP (relative standard errors of prediction) less than 1.176 and 15.5% respectively. The results suggest that PSO-LS-SVM algorithm has a good model performance and high prediction accuracy. NIR has a potential value for rapid determination of multi-indicators in Corni Fructus.

Determination of four effective components from total flavonoids of Scutellaria barbata by high performance liquid chromatography / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish a HPLC method for simultaneous determination of 4 effective components from total flavonoids of Scutellaria barbata (FSB).</p><p><b>METHODS</b>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.</p><p><b>RESULT</b>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.</p><p><b>CONCLUSION</b>The method can be used effectively to evaluate the quality of FSB.</p>

Comprehensive quality control method for total flavonoid of Fructus Aurantii / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish a comprehensive quality control method for total flavonoid of Fructus Aurantii.</p><p><b>METHODS</b>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.</p><p><b>RESULT</b>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.</p><p><b>CONCLUSION</b>The method reported in this paper can be used effectively for the quality control of total flavonoid of Fructus Aurantii.</p>

Influence of H2O2 on degradation of residual pesticides and constituents in Radix Sophorae Flavescentis / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the characteristic and influential factors of the degradation of residual pesticides and alkaloids in Radix Sophorae Flavescentis by H2O2.</p><p><b>METHOD</b>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.</p><p><b>RESULT</b>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.</p><p><b>CONCLUSION</b>H2O2 is a useful reagent for the degradation of organophosphorus and pyrethroid in crude drug.</p>

Effect of griffithin on anticancer activity and apoptosis of cancer cells in vitro / 药学学报

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.

The effect of preparation method on the quality of shen-mai injection / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of different preparation method on the quality of Shen-mai injection.</p><p><b>METHOD</b>The Shen-mai injection samples were prepared using three different methods. Fingerprints of Shen-mai extracts red ginseng, and its intermediates were obtained using an HPLC analytical procedure. The contents of ginsenoside Rg1, Rc and Rb1, and the gross saponins of Shen-mai extract were quantitatively mensured with HPLC procedures.</p><p><b>RESULT</b>There was significant difference in fingerprints and chemical contents of the injections prepared by the three different methods.</p><p><b>CONCLUSION</b>The quality of Shen-mai injection was greatly influenced by the preparation method. HPLC fingerprinting method can be applied for the determination of the Shen-mai preparations.</p>

A novel fingerprint method for quality evaluation of Chinese medicinal plants based on analytical data visualization / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate new method for evaluating the quality of Chinese Medicinal Plants (CMP).</p><p><b>METHOD</b>A visualization technique for representing instrumental analytical data was developed by applying the fundamental of Data Visualization, with Principal Component Analysis (PCA) and spatial projection transformation, original IR spectral data were projected into a low-dimensional subspace so that the dimensionality of original data space was decreased and tiny fingerprint features were extracted. The data set in the subspace was visualized by means of two-dimensional grayscale images. Consequently, the characteristic fingerprint for appraising the quality of CMP was obtained.</p><p><b>RESULT</b>42 mulberry root-bark samples from three different quality classes were identified with the proposed method, which showed that the fingerprint images had satisfactory resolution and classification accuracy as high as 90.5%.</p><p><b>CONCLUSION</b>The proposed method is a useful technique for appraising the quality of CMP.</p>

Simultaneous determination of 4 anthraquinones in Kuhuang injection by RP-HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To develop a new high-performance liquid chromatographic method for simultaneous quantitative determination of 4 anthraquinones in Kuhuang injection.</p><p><b>METHOD</b>The chromatographic separation was performed on a Lichrospher C18 column (4.6 mm x 250 mm, 5 microm,), A linear gradient elution of A (CH3COOH: H2O = 1:100) and B (CH3COOH: H2O: CH3CN = 1:20:80) was used. The flow rate was 0.8 mL x min(-1) and column temperature was set at 35 degrees C. The UV detection wavelength was set at 254 nm.</p><p><b>RESULT</b>The recoveries of rhein, emodin, chrysophanol and physcion were 98.9%, 100.5%, 102.5% and 99.0%, respectively. Their linear ranges were 0.0875-1.75 microg for rhein, 0.0825-1.65 microg for emodin, 0.159-3.17 microg for chrysophanol and 0.0525-1.05 microg for physcion, respectively.</p><p><b>CONCLUSION</b>The method was accurate, repeatable and suitable to determine the contents of 4 anthraquinones in Kuhuang injection.</p>

Study on the solubility changes of Rhizoma Coptidis' main chemical constituents influenced by the proportion of Rhizoma Coptidis and Fructus Evodiae / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the relationship between the solubility of Rhizoma Coptidis' main chemical constituents and the proportion of Rhizoma Coptidis and Fructus Evodiae.</p><p><b>METHOD</b>9 samples were selected in which the proportion of Rhizoma Coptidis and Fructus Evodiae were 1:0, 6:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:6 and 0:1, respectively. The samples' water-extracts were analyzed by RP-HPLC. Then solubility changes of eight chemical constituents were calculated, and the regression equations were established.</p><p><b>RESULT</b>Among the eight chemical constituents, one was independent on the change of proportion of Rhizoma Coptidis and Fructus Evodiae, six having linear relationship. A new constituent was detected, and there was nonlinear relationship between its quantity and the proportion of the couple.</p><p><b>CONCLUSION</b>The solubility of Rhizoma Coptidis' main chemical constituents declines almost in the same scale when Rhizoma Coptidis and Fructus Evodiae are used as a couple, and a new constituent has been detected which can't be found in the mono-extraction.</p>

Determination of amlodipine in CYP3A4 cDNA-expressed cells by HPLC / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish a RP-HPLC method for the determination of amlodipine after metabolism by cytochrome P450 cDNA-expressed cells.</p><p><b>METHODS</b>The determination was performed on a C(18) reversed phase column with a mobile phase composed of acetonitrile phosphates buffer (45:55, v/v, pH 4.5) with UV detection (lambda250nm). Propranolol was used as the internal standard.</p><p><b>RESULT</b>The standard curve was linear over the concentration range of 0.2 - 30.0 microg/ml (r=0.9993), and the limits of determination was 20 ng/ml (S/N >or=3), the limits of quantity was 0.2 microg/ml (recovery 104.0%, RSD 11.4%, n=5). The recovery for this assay was (98.2+/-2.4)%, precision for inter-assay and intra-assay was <10 % and 6 %, respectively.</p><p><b>CONCLUSION</b>The HPLC method established is simple, accurate and suitable for the determination of amlodipine in cytochrome p450 cDNA-expressed cells.</p>

Identification of the habitat of Ligusticum chuangxiong with chromatographic fingerprinting / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the chromatographic fingerprinting for identifying the habitat of Ligusticum chuangxiong.</p><p><b>METHOD</b>HPLC system was applied to obtain the chromatograms of L. chuangxiong samples from different areas, and 15 peaks were measured from the chromatograms. Then some computer-based methods including principle component analysis, clustering analysis, similarity calculation and fisher factor analysis were applied for data analysis.</p><p><b>RESULT</b>There was obvious difference among chromatographic fingerprints of L. chuangxiong samples from different areas. The 15 measured peaks could be used as the fingerprint features.</p><p><b>CONCLUSION</b>Chromatographic fingerprinting can be used for identifying the habitat of L. chuangxiong.</p>