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OBJECTI VE To establish the high performan ce liquid c hromatography（HPLC）fingerprint of carotenoid in Lycium barbarum，and to investigate the spectrum-effect relationship between its common peak and antioxidant activity. METHODS HPLC method was adopted. The fingerprints of carotenoid in 34 batches of L. barbarum from different producing areas were established by Similarity Evaluation System of TCM Fingerprint （2012 edition），and similarity evaluation and common peak identification were carried out. Taking scavenging rate of DPPH free radical as index ，in vitro antioxidant activity of carotenoid in L. barbarum was investigated. The spectrum-effect relationship between the common peaks of carotenoids in L. barbarum and antioxidant activity was analyzed by grey correlation method. RESULTS There were 4 common peaks in the fingerprints of carotenoids in 34 batches of L. barbarum ，and the similarity was not less than 0.903. Peak 1 was identified as zeaxanthin ，and peak 4 as zeaxanthin dipalmitate. The scavenging rates of them to DPPH free radical were 1.792％-3.160％. The common peaks of carotenoids in L. barbarum were positively correlated with scavenging rate of DPPH free radical ，and the correlation degree was greater than 0.6；the correlation degree of peak 2 and peak 4（zeaxanthin dipalmitate ）with scavenging rate of DPPH free radical was greater than 0.8. According to the correlation degree ，the contribution of each common peak to scavenging rate of DPPH free radical was determined as peak 2＞ peak 4（zeaxanthin dipalmitate ）＞peak 1（zeaxanthin）＞peak 3. CONCLUSIONS In this study ，HPLC fingerprint of carotenoid in L. barbarum is successfully established ，and two common peaks are identified. The chemical components represented by peak 2 and zeaxanthin palmitate may be the material basis of antioxidant activity of carotenoid in L. barbarum .
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Objective:To establish an HPLC method to determine the entrapment efficiency (EE) and drug loading (DL) of curcumin (CUR)and quercetin (QUE)loaded self-microemulsifying drug delivery system.Methods:A centrifugation method was used to isolate the free drug.The content of drug was determined by HPLC.The analytical column was a Purospher STAR LP C18 column (250 mm×4.6 mm,5 μm) and the column temperature was 30 ℃.The mobile phase was acetonitrile-4% acetic acid (50∶50) and the flow rate was 1.0 ml·min-1.The UV detection wavelength was set at 370 nm and the injection volume was 10 μl.Results:CUR and QUE were linear within the range of 10.728-96.552 μg·ml-1 (r=0.999 8) and 1.08-9.72 μg·ml-1 (r=0.999 9),respectively.The average recovery was 99.98%(RSD=1.46%,n=9) and 100.34%(RSD=1.06%,n=9),respectively.In CUR-QUE-SMEDDS,the EE of curcumin and quercetin was (95.97±0.50)% and (95.91±2.52)%,and the DL was (25.82±0.15) mg·g-1 and (1.80±0.05)mg·g-1,respectively.Conclusion:The method is accurate,rapid and simple,and suitable for the determination of DL and EE in CUR-QUE-SMEDDS.
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The aim of the study was to prepare and evaluate the quality of negatively charged selfmicroemulsifying drug delivery system of curcumin (NC-CUR-SMEDDS).Based on the CUR-SMEDDS,the optimum amount of excipients was confirmed by the single-factor design experiment taking mean particle size,Zeta potential,drug entrapment efficiency and the drug loadings of curcumin as the evaluation indices for the preparation of NC-CUR-SMEDDS.The quality of NC-CUR-SMEDDS was evaluated by observing its appearance status,transmission electron microscope micrographs and determining particle diameter,Zeta electric potential,drug entrapment efficiency and drug loading.As a result,it was found that the Zeta potential reached -43.43 ± 0.29 mV when 4％ docusate sodium was added.The appearance of NC-CUR-SMEDDS remained clarified and transparent,and the microemulsion droplets appeared spherical without aggregation with uniform particle size distribution.The mean particle size was 14.08 ± 0.082 nm,the drug loading of curcumin was 26.48 mg·g-t and the drug entrapment efficiency was 94.12％.It was concluded that NC-CUR-SMEDDS with high entrapment efficiency and uniform particle size distribution met the requirement of inflammatory target binding in the colon.
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This study was aimed to explore differential effects of various sections of the velvet antler on promoting cell proliferation in vitro. The NRK-49F cell line from rat kidney fibroblasts was used as the cell model. The cell proliferation rates of the water extracts from the upper, middle and lower section of fresh velvet antler were measured by the MTT method. BCA method was used in the detection of protein concentration. The SDS-PAGE method was used in the analysis of difference composition of the sample protein. The results showed that soluble protein content of the upper, middle and lower section were 17.89, 16.04 and 6.89 mg·mL-1, respectively. From the top to the base, the soluble protein content of velvet antler was decreased. After 24 h treatment, when the protein concentration of the upper and middle section samples of the velvet antler were 800 μg·mL-1 and 600 μg·mL-1, the cell proliferation promoting rates reached the maximum, which were 66.76% and 64.36%, respectively. And when the lower section sample of the velvet antler was 1 000 μg·mL-1, the cell proliferation promoting rates reached the maximum, which was 58.87%. After 48 h treatment, when the upper and middle section samples of the velvet antler were 800μg·mL-1, the cell proliferation promoting rates reached the maximum, which were 219.56% and 215.86%, respectively. And when the lower section sample of the velvet antler was 1 000 μg·mL-1, the cell proliferation promoting rates reached the maximum, which was 169.20%. The velvet antler on the proliferation of cells was much better than the 10% fetal bovine serum. The figure of SDS-PAGE showed the slight difference in the protein composition of three part of the velvet antler. It was concluded that all samples had promoting effects on cell proliferation with concentration-depen-dent, and the main protein in different part of the velvet antler had minor differences.
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Objective To establish the fingerprints for characterization of the chemical components of maidong(root of Ophiopogon japonicus) in two main cultivate regions of China,Sichuan(Chuanmaidong) and Zhejiang(Hangmaidong).Methods An HPLC-UV analytical methods was applied to detect 70% ethanol extracts of 20 samples from Sichuan and Zhejiang province,a "Fingerprint similarity evaluating system for TCM" issued by Pharmacopoeia Committee of P.R.China was used to evaluate the similarities all of the samples.Results The fingerprints revealed that the similarities were higher than 0.95 between samples from the same cultivate region,and were lower than 0.80 between samples from different regions of above two.Conclusion The fingerprints of Chuanmaidong and Hangmaidong were provided with high difference,and the difference can be taken as a most important proof for distinguishing the material medica of Chuanmaidong and Hangmaidong,but also in patent medicine of TCM.
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Objective To determine the contents of methylophiopogonanone A (MOA) and methylophiopogonanone B (MOB) in Radix Ophiopogonis and its extracts. Methods An HPLC-UV method was used for determining the contents of MOA and MOB in all samples. Analytical column was Kromasil C18 (250 mm?4.6 mm, 5 ?m). Mobile phase was acetonitrle-water (55∶45) and detection wavelength was 298 nm. The flow rate of mobile phase was 1 mL/min, and temperature was 30 ℃. Results The contents of MOA in Radix Ophiopogonis cropped in Sichuan and Zhejiang Provinces were 0.004 0%-0.009 6%, 0.006 7%-0.013 4%, and the contents of MOB were 0.002 1%-0.006 2%, 0.015 9%-0.028 2%, respectively. The contents of MOA in the extract of Radix Ophiopogonis cropped in Sichuan and Zhejiang Provinces were 0.007 5%-0.008 8%, 0.011 2%-0.012 6%, and the contents of MOB were 0.003 8%-0.005 1%, 0.020 7%-0.023 8%, respectively. Conclusion The contents of MOA and MOB in Radix Ophiopogonis cropped in Zhejiang Province and its extracts are more than those in Sichuan Province and its extrouts. The method can be used for the purpose of the quality control of Radix Ophiopogonis and its extracts.