| The quality of Traditional Chinese Medicine (TCM) determined the efficacy and clinical effect. How to gain high quality medicinal material has been the focus of researcher attention. Chuanxiong, the dried rhizome of L. chuanxiong Hort. (Umbelliferae), is a herb that has been widely used in TCM for a long time. It is frequently prescribed to treat angina pectoris, cardiac arrhythmias, hypertension, and stroke because it helps in blood circulation and disperses blood stasis. In this manuscript, L. chuanxiong as the research object, firstly, author build the HPLC fingerprint, high-pressure ultrasonic optimization extraction new technology, and artificial neural network quality quantitative evaluation mathematical model; secondly, through to system research the NPK ratio, special fertilizer, plant density, sowing date and harvesting date, and formulated the L. chuanxiong standardized cultivation of standard operation procedure. The main results were as follows:1. Establishment of HPLC fingerprintDujiangyan, pengzhou, pengshan, and shifang of L. chuanxiong samples for research material, author investigated the six major constituents under different determination conditions, single factor tests were employed to determine the mobile phase, flow rate, column temperature, and wavelength. The mobile phase consisted of acetonitrile (A), methanol (B), and 1% aqueous acetic acid (C) using a gradient program of 20:40:40 (A:B:C, v/v/v) at 0 min to 5 min,60% to 100%(A), and 40% to 0%(C) from 5 min to 30 min. The flow rate was 0.6 mL·min-1, and the column temperature was 25℃. The detection wavelength was set to 0 min to 7.3 min 321nm, 7.4 min to 20.8 min 275nm,20.9 min to 23 min 260 nm, and to 275 nm from 23.1 min to 30 min. The HPLC fingerprint method, linearity, precision, accuracy, stability, and recovery are all comply with the relevant requirement of chromatographic fingerprint. To qualitative analyzed the common peak of fingerprint chromatographic, and identified ferulic acid, senkyunolide I, senkyunolide H, senkyunolide A, ligustilide, and levistilide A.2. Establishment of high-pressure ultrasound-assisted extraction new technologyHigh-pressure ultrasound-assisted extraction technology was applied to extract ferulic acid, senkyunolide I, senkyunolide H, senkyunolide A, ligustilide and levistilide A from L. chuanxiong rhizomes. Seven independent variables, including solvent type, pressure, particle size, liquid-to-solid ratio, extraction temperature, ultrasound power, and extraction time were examined. Response Surface Methodology (RSM) using a Central Composite Design (CCD) was employed to optimize the experimental conditions (extraction temperature, ultrasonic power, and extraction time) on the basis of the results of single factor tests for the extraction of these six major components in L. chuanxiong rhizomes. The experimental data were fitted to a second-order polynomial equation using multiple regression analysis and were also examined using appropriate statistical methods. The best extraction conditions were as follows:extraction solvent:40% ethanol; pressure:10 MPa; particle size:80 mesh; liquid-to-solid ratio:100:1; extraction temperature:70℃; ultrasonic power,180 W; and extraction time,74 min. In this study, seven factors, namely solvent type, pressure, particle size, liquid-to-solid ratio, extraction temperature, ultrasound power, and extraction time, were investigated first to optimize the extraction solvent, pressure, particle size, and liquid-to-solid ratio. Additionally, the levels of the response surface experimental design factors (temperature, power, and time) were determined according to the extraction temperature, ultrasonic power, and extraction time optimized in the single factor tests.3. Establishment of quantitative quality evaluation methodTo the above 6 ingredients content of L. chuanxiong for quality evaluation index, and first proposed using quality evaluation index (δ, is the dimensionless parameter for quantitative description of medicine quality) for quantitative evaluation of TCM. Using the developed HPLC fingerprint method to analyze the samples, and obtained data to through the principle components analysis (PCA), hierarchical cluster analysis (HCA), similarity evaluation (SE), and quality evaluation index (δ) analyzed. The artificial neural network (ANN) quantitative quality evaluation model of L. chuanxiong, as shown below: H1=tanh(0.5(10.6066x1-1.3179x2+114001x3+1.6723x4+0.5779x5-26.2736x6-5.6332)) H2=tanh(0.5(13.2457x1-9.1249x2+11.6289x3-0.1441x4+0.1966x5-34.2369x6-4.8899) H3=tanh(0.5 (-2.0291x1+19.4490x2-5.0848x3+0.5648x4+0.2370x5+49.9950x6-4.0533 δ=0.0708H1+0.2400H2+0.2322H3+0.90214. Formulated the L. chuanxiong standardized production of operation procedureThrough to system research effects of NPK ratio, special fertilizer, plant density, sowing date, and harvesting date on the quality and yield of L. chuanxiong, determination the N 201～206 kg·hm-2, P 458～522 kg·hm-2, and K 362～388 kg·hm-2; plant density 160000～180000 hills·hm-2; the best sowing data and harvesting date are 8.1 to 8.5 and 5.20 to 5.25, respectively. |
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