Orthogonal experiments to optimize the extraction of tanshinone ⅡA
Tanshinone is a fat-soluble diterpenoid compound with orange-yellow and orange-red features in Salvia miltiorrhiza, which can be classified into more than 10 kinds of components according to its structure, such as tanshinone I, tanshinone IIA, tanshinone IIB, cryptotanshinone, dihydrotanshinone I, hydroxytanshinone, tanshinone methyl, isotanshinone, isocryptotanshinone, etc., among which, tanshinone IIA is the representative of the fat-soluble components of Salvia miltiorrhiza, and it has natural antioxidant effects. Tanshinone IIA has a natural antioxidant effect, and a large number of studies have shown that it has good therapeutic effects in anti-tumor, cardio-cerebral and cerebral vascular diseases, antibacterial and anti-inflammatory effects, and so on.
Operation method
orthogonal method
Principle
1. Tanshinone IIA (Tanshinone IIA) is derived from the dried roots and rhizomes of Salvia miltiorrhiza Bge. in the family Labiatae, the roots of Salvia sclarea L. in southern Europe, and the roots of Salvia sclarea L. or Salvia officinalis in Longxi. Orange-red needle-like crystals (EtOAc), mp 209～210 ℃. Easily soluble in ethanol, acetone, ether, benzene and other organic solvents, slightly soluble in water. 2. Principle of orthogonal experimental design Orthogonal experimental design (Orthogonal experimental design) is the study of multifactorial and multilevel and another design method, which is based on the orthogonality of the full test from the test to select some representative points for the test, these representative points have a " These representative points have the characteristics of "uniform dispersion and comparability", orthogonal experimental design is the main method of fractional factorial design. Orthogonal experimental design is the main method of fractional factorial design. It is a kind of experimental design method with high efficiency, rapidity and economy. Japanese famous statistician Genichi Taguchi will orthogonal test selected level combinations into a table, called orthogonal table. For example, to make a three-factor three-level experiment, according to the comprehensive experimental requirements of Xiamen University School of Medicine, Department of Pharmacy - Natural Medicinal Chemistry Laboratory Handout, 33 = 27 combinations of experiments, and has not yet considered the number of replicates of each combination. If the experiments are arranged according to L9(33) orthogonal table, only 9 times need to be made, if L18(37) orthogonal table then 18 experiments are carried out, obviously greatly reducing the workload. Thus orthogonal experimental design has been widely used in many areas of research. Orthogonal table is a set of rules for the design of the table, using L as the code name of the orthogonal table, n is the number of experiments, t is the number of levels, and c is the number of columns, that is, the number of factors that may be arranged to maximize the number of factors. For example, L9(34), which indicates that 9 experiments are required, with a maximum of 4 factors to be observed, each at 3 levels.
Materials and Instruments
Salvia divinorum crude powder Move 1. Extraction methods For more product details, please visit Aladdin Scientific website.
Water
Volumetric instrument Extraction filter Microporous filter membrane HPLC
The crude powder of Salvia miltiorrhiza was weighed 2.5 g, and the experiments were arranged according to the orthogonal test table L9 ( 33 ). After immersing the herbs for 30 min, the extract was extracted by ultrasonic sound, and the filtrate was collected by filtration, and the volume of the filtrate was measured. The filtrate was collected by filtration, and the volume of filtrate was measured. 0.25 ml of the filtrate was accurately aspirated into a 25 ml volumetric flask, diluted with methanol, shaken well, and filtered through a microporous membrane, and then determined by HPLC.
2. Orthogonal design
According to the physicochemical properties of tanshinone IIA, three influencing factors were selected in the extraction, and the three-factor level orthogonal test table L9 ( 33 ) was used to design the experimental program, see the following table.
3. Orthogonal experimental data processing: refer to the following table processing method to select the optimal extraction method with the purity and transfer rate as indicators.
