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Objective: To established a rapid nondestructive determination method for the multi-marker constituents in Angelica sinensis by near infrared spectroscopy (NIRS) combined with the partial least squares (PLS) method in order to improve the quality control for A. sinensis. Methods: A total of 108 batches of A. sinensis from different origins were collected for the research. An Ultra performance liquid chromatography (UPLC) method was established to measure the content of the seven components in A. sinensis, which were adopted as the reference value. And the integrating sphere diffuse reflection mode was employed to collect the NIR spectrum. The quantitative calibration model between the near infrared spectrum and the reference content of each component to be measured was established by PLS and other chemometrics methods. Each part of the modeling process was optimized respectively, including the selection of calibration set and validation set, different pretreatment methods and different spectral section. Results: The correlation coefficient for calibration set of chlorogenic acid, ferulic acid, isochlorogenic acid A, ligustilide, butylidenephthalide, senkyunolide I and levistilide A were 0.937 6, 0.970 2, 0.963 4, 0.991 1, 0.962 4, 0.966 6 and 0.947 6, respectively; The root mean square error of prediction (RMSEP) were 0.072 1, 0.038 9, 0.011 3, 0.483 0, 0.017 5, 0.178 0 and 0.097 0, respectively. The predicted values of NIRS models and the measured values of UPLC showed a good linear relation, which presented a great prediction ability of the models. Conclusion: The methods of NIRS combined with PLS can be applied for the rapid content determination of seven components in A. sinensis including chlorogenic acid, ferulic acid, isochlorogenic acid A, ligustilide, butylidenephthalide, senkyunolide I and levistilide A, which is proved to be simple and reliable.
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Objective To establish a quantitative analysis method of multiple active components (gallic acid, tanshinol, ferulic acid, rosmarinic acid, salvianolic acid B, luteolin, apigenin, aloe-emodin, emodin, butylidenephthalide, and levistilide A) in Fukejing Capsule (FC) based on ultra performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UPLC-Q- Orbitrap HRMS). Methods The column was BEH C18 (50 mm × 2.1 mm, 1.7 μm) and the mobile phase was consisted of acetonitrile-water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min with gradient elution; Mass spectrometer conditions: Heated electrospray ionization source (HESI) and full mass quantify methods were used to perform the determination, and the results of the contents were imported into the muti-data processing soft ware SIMCA14.0 to make a quality evaluation. Results Under the optimized conditions, gallic acid, tanshinol, ferulic acid, rosmarinic acid, salvianolic acid B, luteolin, apigenin, aloe-emodin, emodin, butylidenephthalide, and levistilide A all showed good liner relationship (r ≥ 0.999 5) in the range of 0.15—1.50, 0.80—8.00, 1.50—15.00, 0.04—0.40, 0.015—0.150, 0.10—1.00, 0.004—0.040, 0.004—0.040, 0.02—0.20, 0.01—0.10, and 0.04—0.40 μg/mL, respectively; The results of the accuracy, the repeatability and the stability all reached the standards (RSD ≤ 4%); The recoveries ranged from 98%—102% and RSDs were below 3%; The result of the content ranges in different batches were 26.36—31.03 μg/g (gallic acid), 178.85—210.79 μg/g (tanshinol), 320.91—343.16 μg/g (ferulic acid), 2.84—3.09 μg/g (rosmarinic acid), 3.55—4.25 μg/g (salvianolic acid B), 27.33—32.36 μg/g (luteolin), 1.89—2.13 μg/g (apigenin) A, 0.47—0.60 μg/g (aloe-emodin), 3.17—3.57 μg/g (emodin), 1.99—2.54 μg/g (butylidenephthalide), and 7.51—8.53 μg/g (levistilide A). The analysis results showed that the quality of the most batches was stable, the tanshinol and ferulic acid had a great influence on the quality of the medicine, and could be monitored to ensure the quality of different batches. Conclusion The methods established in this paper have a high sensitivity and accuracy; The results of the methodology conform to the relevant requirements and the methods can rapidly determinate the multiple active components in FC; The research also provides a new scientific basis and reference for the quality assessment at the same time.
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Objective: To optimize the processing technology for prepared Chuanxiong Rhizoma by D-optimal response surface methodology with comprehensive weighted of multi-index. Methods: The single factor experiment was used to optimize the dilution ratio and the moistened time of rice wine. Then, based on the contents of five main components (ferulic acid, ligustilide, senkyunolide A, senkyunolide I, and 3-butylidenephthalide), the D-optimal response surface methodology was used to optimize the rice wine dosage, stir-fried temperature, and stir-fried time in the processing technology for prepared Chuanxiong Rhizoma. Results: The optimal processing technology for prepared Chuanxiong Rhizoma was mixed with 20% rice wine, moistened for 75 min, and stir-fried for 12 min at 156 ℃. Also, it was found that the contents of senkyunolide A, senkyunolide I, and 3-butylidenephthalide in prepared Chuanxiong Rhizoma had increased, and it should be associated with processing. Conclusion: The processing technology for prepared Chuanxiong Rhizoma optimized by D-optimal response surface methodology with comprehensive weighted of multi-index can be used to optimize the process parameters, promote the stability and repeatability of the processing technology, which can significantly control the inner quality of prepared Chuanxiong Rhizoma to ensure effectiveness in clinical application.
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Objective: This current study focused on the identification of active constituents from Angelica Sinensis Radix in Xiaoyao Powder based on UPLC-PDA-guided isolation technique. Methods: The UPLC-PDA chromatogram of Xiaoyao Powder was compared with that of Angelica Sinensis Radix. The relative retention time of each peak and the Uhraviolet spectra provided by PDA were used in the analyses. The constituents were isolated from Angelica Sinensis Radix under the guidance of UPLC-PDA investigation. The structures of the isolates were elucidated by NMR techniques. The antidepression effect was evaluated on glutamate-induced neurons. Results: Five marker peaks of Xiaoyao Powder fingerprint belonged to Angelica Sinensis Radix and they were determined as coniferyl ferulate (1), E-butylidenephthalide (2), ligustilide (3), Z-butylidenephthalide (4), and 14-acetoxy-12-senecioyloxytetradeca-2E,8E,10E-trien-4,6-diyn-1-ol (5). Compound 5 was isolated from the plants in Umbelliferae for the first time. The treatment with compounds 1, 3, and 4 could protect PC12 and SH-SY5Y cells from glutamate-induced cytotoxicity. Antidepression bioactivity of compound 1 was first investigated. Conclusion: UPLC-PDA-guided isolation technique is confirmed to be a rapid and accurate method to identify the main active constituents from Angelica Sinensis Radix in Xiaoyao Powder.
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Aim To investigate the bio-affinities of ligustilide and butylidenephthalide to rat aortic smooth muscle cells and the inhibitory effects of them on bFGF-stimulated proliferation of rat vascular smooth muscle cell (VSMC). Methods VSMCs were cultured from rat aorta pectoralis and identified by an immunohistochemical method. The bio-affinities between solute (ligustilide or butylidenephthalide) and cell membrane were measured by rat aortic cell membrane chromatography (CMC). The inhibitory effects of ligustilide and butylidenephthalide on bFGF-stimulated VSMC proliferation were evaluated by MTT colorimetric method. Results Both ligustilide and butylidenephthalide had selective affinities to rat aortic smooth muscle cell as the same as verapamil, one of the calcium ion antagonists. They could potently separately (P ＜ 0.05 ), but had no effects on the normal VSMC growth. Conclusion Both ligustilide and butylidenephthalide can inhibit the abnormal proliferation of VSMC induced by bFGF.