Extraction, separation and identification of the composition of the jaundium in the yellow.

The
the Great Yellow is recorded in many literatures, such as the Book of God's Agricultural Herbs, and is used for leakage, stomach health, heat clearing, detoxification, etc.
since ancient times, the great yellow in the plant-based laxative medicine occupies an important position, is a very early by the national pharmacopeia contained in the world's raw medicine. There is a wide variety of large yellows, and the high-quality large yellows are Rheum palmatclm L, and the big yellows (R. Officinale Baill) and Tanggut Huang (R. Tangutium Maxim.et Regll) roots and roots, and the large yellow contains a variety of free hydroxyquinone compounds and the glycosides they form with sugar. There are five main types of hydroxyquinone that have been known:
large yellow mid-yellow quinines, which have different structures and therefore different acidity. Large yellow acid is connected with -COOH, the most acidic, big lutein is connected with β-OH, acidity is second, aloe vera big yellow is connected with phenol-OH, acidity is third, large yellow methyl ether and large yellow phenol are 1,8-diphenol hydroxyl, the former is linked to -OCH3 and -CH3, the latter is only linked to CH3, so the latter acidity ranks fourth.the purpose and requirements of the experiment
1. Learn the basic operating
of
of the buffer paper and design the experimental scheme of liquid extraction to separate the mixture according to the chromatography results.
2. Master the principle and operation technique of PH gradient method.
3. The test of the separation of large yellow phenol and large yellow methyl ether by calcium phosphate column chromatography is further familiar with the column chromatography operation technique.
4. Learn how to identify quinine compounds.experimental method
1. Extraction of large yellow total quinine elements
Note: (1) Most of the quinine components in the large yellow are combined with sugar and are present in plant
tissues in the form of
. So use acid hydrolysing to produce nucleotides. Phenyl glycosides can be dissolved in chloroform, benzene and ether and other organic solvents, with benzene should pay attention to the volatile benzene vapor, strictly prevent poisoning; Chloroform extraction in the placement of precipitation, can be filtered out, the precipitation is mostly large yellow, residual liquid for the next separation test.
2. The design of the total quinine element separation scheme
1) the paper chromatography
2) the buffer paper chromatography test
3) the best extractor and its dosage determination
4) the determination of the various extractor dosages of the various extraction agents
5) the design of the new separation method
3. Separation of quinine components and refined
(1) separation and refining of jaundate
will contain total free chloroform 450ml to 1000ml of large sub-liquid funnel, plus PH8 buffer 275ml vibration extraction, rest to complete layering, release chloroform, to the alkaline water solution to 250ml
0 ml In the
smooth" , add HCl acidification to PH 3, after the yellow precipitation is fully precipitated,
filter
,
dry
, dry sample with ice acetic acid 10 ml
heating
to make the solution, while heat filtration, filter fluid static, the yellow needle crystal for large yellow acid, filtration is pure.
(2) Separation and refining of large lutein
will continue to move chloroform with too much yellow acid into the split funnel, with PH9.9 buffer 275 ml shake extraction, thoroughly layered, the division of alkaline water Layer, and with HCl acidification to PH-3, precipitated brown-yellow precipitation, filtration, precipitation after drying, with 10 ml of ice acetic acid heating to dissolve, while hot filtration, analysis of orange needle crystal, filtration, that is, large jaundrine pure.
(3) separation and refining of aloe vera large lutein
residual chloroform liquid to the split funnel, plus 5% NaCO3:5% NaOH (9:1) alkaline ahydrate 540 ml or with 0.5% KOH540ml shaking extract, alkaline water liquid plus HCl acidification, precipitated precipitation filter drying, with 10 ml ethyl acetate recrystrystry, yellow needle crystal aloe vera large lutein pure.
(4) separation of large yellow phenol and large yellow methyl ether - calcium phosphate column
4. Identification of steroid ingredients
1) Paper chromatography
2)
IR
spectral
parsing The main points and notes of this experiment
1) pH gradient extraction principles and precautions.
2) How to design the procedures and methods of extraction separation schemes.
3) the characteristics of the infrared spectrum of the compound.
4) When separating extraction, be sure to pay attention to the separation of emulsified layers, do not mix in, and each step is best with fresh CHCl3, back wash alkali water.
5) The preparation of the buffer and the preparation of the lye should be accurate, pay close attention to the inspection.
6) At each step of extraction, use paper chromatography to check the tracking extraction effect, if the desired effect is not achieved should be corrected in a timely manner.
.