Method of quickly screening standard substance used in quantitative detection of trifolium pratense isoflavone
A technology for the quantitative detection of red clover isoflavones, which is applied in the field of rapid reduction and screening of standard products used in the quantitative detection of red clover isoflavones, can solve problems affecting the development of the red clover industry and achieve low detection costs and fast screening , The effect of simple process
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Embodiment 1
[0015] Example 1 Effect of Different Standard Items on Spectrophotometric Determination of Red Clover Isoflavones Results
[0016] 1. Preparation of red clover samples: Three red clover samples were randomly collected, air-dried and pulverized, and named samples 1, 2 and 4. Take 0.2 g of naturally air-dried red clover leaves, soak them in 8 mL of methanol, and extract them by ultrasonic wave for 30 minutes. After centrifugation for 10 minutes, the supernatant was taken and filtered through a 0.45 micron organic membrane to prepare the test solution.
[0017] 2. Preparation of standard products:
[0018] Weigh 5mg of formononetin reference substance and place it in a 100mL volumetric flask, add methanol to dilute to the mark to obtain a formononetin reference substance solution with a concentration of 0.05mg / mL.
[0019] Weigh 10mg of genistein standard substance into a 100mL volumetric flask, and then add methanol to calibrate to the scale to obtain a genistein standard solu...
Embodiment 2
[0030] Embodiment 2 Thin-layer chromatography screening standard method
[0031] Using a capillary tube, spot the mixed samples of the four standard products and the samples of red clover 1, 2 and 4 to be tested on the prepared silica gel GF254 thin-layer chromatography plate, put them into a developing bottle, and use chloroform:methanol= 15:1 is the developing agent for developing. When the developing agent reaches 1-2 mm from the edge of the TLC plate, take out the chromatographic plate, dry it, and observe it under a 254nm ultraviolet lamp.
[0032] Such as figure 2 As shown, in the figure B represents garbanzoin A; F represents formononetin; G represents genistein; D represents daidzein. The spots of samples 1 and 2 formononetin (standard product F) are more obvious, while the spots of sample 4 chickpein A (standard product B) are more obvious.
[0033] Based on the results, it is suggested that samples 1 and 2 use formononetin or daidzein as the standard, and sample ...
Embodiment 3
[0034] Embodiment 3 Thin-layer chromatography screens standard substance method
[0035] Because the standard substances chickpein A and formononetin have strong ultraviolet absorption, only these two standard substances can be used to complete the screening.
[0036] Using a capillary tube, the mixed sample mixed with the standard chickpein A and formononetin and the red clover 1, 2 and 4 samples to be tested were spotted on the prepared silica gel GF254 thin-layer chromatography plate to Chloroform:methanol=20:1 for development. When the developing agent reaches 1-2 mm from the edge of the TLC plate, take out the chromatographic plate, dry it, and observe it under a 254nm ultraviolet lamp.
[0037] Such as figure 2 As shown, the spots of formononetin (standard substance F) in samples 1 and 2 are more obvious, while the spots of chickpein A (standard substance B) in sample 4 are more obvious.
[0038] Based on the results, it is suggested that samples 1 and 2 use formonon...
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