Key words: Schizonepeta
; total flavonoids; spectrophotometry; determination
1 Instruments and materials
TM902C digital thermometer (Shenzhen Haitian Industrial Co., Ltd.); PT-02 thermocouple (Nanjing Thermal Instrument Factory); FA-1004 electronic analytical balance (Shanghai Tianping Instrument Factory); UV-2401 UV-visible spectrophotometer (Japan) Shimadzu).
1.2 The material of schisandra chinensis was purchased from Hebei, Shandong and Jiangsu, and was identified as the spike of the schizonepeta plant of the Labiatae family by Professor Wu Qinan of the School of Chinese Medicine Identification. Wash and cut into small pieces for use. The self-broiled mustard charcoal system will be stalked with stalks, and the charcoal method will be used to control the temperature of 220 Â° C for 8 min. The rutin
reference substance was purchased from the China National Institute for the Control of Pharmaceutical and Biological Products, batch number: 0080-9705. The remaining reagents were of analytical grade.
2 Methods and results
2.1 Preparation of reference substance stock solution Take 120 Â°C under reduced pressure to a constant weight of rutin bismuth product 20mg, accurately weighed, set in a 100mL volumetric flask, add methanol to the mark, shake well, that is (each containing a reed D. 0.2064 mg).
2.2 Preparation of the test solution Take about 2.5g of sample powder (over 20 mesh sieve), accurately weighed, set the Soxhlet extractor, add appropriate amount of petroleum ether, heat and reflux until the extract is colorless, let cool, discard Remove petroleum ether solution, add methanol to the right amount, heat and reflux until the extract is colorless, concentrate, dispose in a 25m1 volumetric flask, wash the container with a small amount of methanol, wash the liquid in a human bottle, add methanol to the mark, shake well, ie Got it.
2.3 Determination of absorption wavelength Take 2 mL of reference substance and 0.5 mL of sample No. 4, respectively, in two 25 mL volumetric flasks, add water to 10 mL, add 1 mL of 5% sodium nitrite solution, mix and place for 6 min (broken vibration) Shake), add 1mL of 10% aluminum nitrate solution, mix, place for 6min (continuous shaking), add 1moL / L sodium hydroxide test solution 10mL. Add water to the mark, shake well, place it for 15 min, scan on the spectrophotometer at 200-800 nn, and determine the detection wavelength to be 500 nm from the scan result.
2.4 Drawing of the standard curve Accurately draw the reference stock solution 0, 2.0, 4.0, 6.0, 8.0 and 10.0mL, respectively, in a 25mL volumetric flask, according to the method of "determination of absorption wavelength", from "add water to Each reference solution was prepared according to law from 10m1". The first solution was blank. The absorbance was measured at a wavelength of 500 nm selected by the spectrophotometer L: the absorbance was plotted on the ordinate and the concentration was plotted on the abscissa, and a standard curve was drawn: the standard curve was: A=1.736+81.93, r=0.9994,
The linear range is from 0.016512 to 0.082560 mg/mL.
2.5 Precision investigation Take the No. 3 reference solution, according to the method under the â€œDrawing of Standard Curveâ€, continuously measure 6 times according to law. The RSD is 0%, indicating that the method has good precision.
2.6 Stability investigation Take 0.5mL of sample solution No. 4 and place it in a 25mL volumetric flask. According to the method under â€œDrawing of Standard Curveâ€, the measurement is carried out every 10 minutes according to the law, and the results show that The stability of the sample solution within 30 min is good, RSD: 2.32% <3% time prolonged, the sample absorbance decreased more: so the selection time is 10-30 min.
2.7 Repeatability test Take sample No. 4 powder, and prepare 6 parts of test solution according to the method under â€œPreparation of test solutionâ€, and accurately draw 0.5mL from 25mL volumetric flask, according to â€œ The method under the â€œdrawing of the standard curveâ€ is determined according to the law from â€œadding water to l0mLâ€. The repeatability results show that the average content of total flavonoids in the sample is 25.58mg/g. RSD = 1.39%.
2.8 Recovery rate Take 6 samples of No. 2 sample, each 1.25 precision weighing, 1 group for 2 groups, a total of 3 groups. Add 25, 31, 38mg of rutin reference substance respectively, and prepare 6 parts of test solution according to the method under â€œPreparation of test solutionâ€, and accurately absorb 0.5m1 in 25mL volumetric flask, according to â€œStandard The drawing of the curve starts from "adding water 10mL" and is determined according to law. The average yield was calculated to be 97.24%, RSD = 1.81%.
2.9 Determination of sample content Precision extraction of the prepared sample of 6 samples of 0.5mL in a 25mL volumetric flask, according to the method of "drawing of the standard curve", from "add water to 10mL" from the law, Calculate the total flavonoid content in the sample.
3 Discussion The extraction methods of total flavonoids in mustard charcoal were: cold soaking, ultrasonic extraction, Soxhlet extraction, etc. The Soxhlet extraction method was used to extract Zui completely.
In this experiment, the content of total flavonoids before and after burning charcoal was compared, and it was found that the content of total flavonoids after burning charcoal was significantly increased. The UV spectra of the schisandra chinensis were compared before and after the charcoal. The results showed that there were significant differences in the UV spectra before and after the charcoal. It was suggested that the content and proportion of flavonoids in the schisandra chinensis were significantly changed before and after charcoal, which may be related to the enhanced hemostatic effect of schisandra chinensis, but the specific composition changes and corresponding proportions need further study.
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