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requirements:
1. The extraction method of steroid saponins (pro-lipid and neutral components) is mastered.
2. Be familiar withand identification of saponins in potatoes.introduction:
potato(Diosgenin) is a type of steroid saponin, molecular form (C
27
H
42
O
31
), molecular weight 414 .61, for white crystallization, mp204 to 207 degrees C, 25D-129.3 (CHCl
3
), dissolved in general
organic
solvent and acetic acid, insoluble in water. Currently, it is an important raw material for the manufacture of a variety of steroidal drugs such as oral contraceptive pills (I, II pills) and steroids
hormones
(e.g. cocoa pine).
In the plant world mainly distributed in the potato dioscorea genus (Dioscorea) plants, China's potatogenus plants have more than 80 species, of which only 17 kinds of potatostem group (Stenophora), I subseeds and a variant contains steroid saponin, others contain polysume, soap-free glycoside. The main ones that have been used for production are D.Zingiberensis C. H. Wright, Dragon Potato(D.nipponica Makino), Yellow Yamant (D.panthaica prain et Burkil), purple turmeric (D.nipponica Makino rosvarth praniin.
in plants,saponins are combined with glucose, rat lily sugar into potatoes(Dioscin) and exist. When extracting separation, the potato suponsine hydrolyzed into potatoessaccharides and monosaccharides (glucose, rat lily sugar) are generally first dissolved with thin acid. Becausesoap glycosides are insoluble in water and mixed in plant residues, organic solvents (such as petroleum ethers) can be used to extract potatosecosides directly from plant residues.extraction method:
1. Saponin pre-test: 2. Extractionof saponins in potatoes:Note 1: Check whether the potatosasines are exhausted, can be used Liberman reaction, reference to experiment II "Note 3" under.
Note II: The method of recovering petroleum ether can be found under "Note 4" of experiment I.
Note 3: The resulting potatosulsine (coarse) is washed with petroleum ether, the melting point can be determined, if the melting point is not qualified, recrystration.identificationsaponins in potatoes:
1. Melting point determination: 204 to 209 degrees C
2. Thin layer identification:
should only present a color spot consistent with the standard RF value.
sample: white crystalline a waterless alcohol liquid . Standard products
: potato sorine-free alcohol liquid
1) adsorbent: AI
2
O
3
soft plate (neutral 100 to 200 eyes, active III.grade)
Expander: Phenyl methanol (9:1)
2) Adsorbent: Silicone H-CMC hard plate
expander: petroleum ether-ethyl acetate (7:3)
colorant: 10% phosphate ethanol solution. After spray
10 to 20 minutes
3) Chemical reaction:
(1) Liebermann-Burchard reaction
(2) CHCI
3
-thick H
2
SO
4
Test
(3) PTA reaction (Kahlenberg reaction): chloroform solution with PTA is purple-blue.
4) Preparation of derivatives:
acetylide method: take samples 100mg dissolved in 3 ml of radon, add 20 ml of acetic anhydride, boil for half an hour to an hour, pour the reaction into ice water (winter operation with cold water). After the white crystal is pre-analyzed, the filtration is extracted and the acetone recrystrystal is available. mp196 to 193 degrees C.
5) UV absorption
spectral
assay:
take samples 5 mg, add thick H
2
SO
4
10ml, heat up in a 40c water bath for 1 hour, cool, assay. Thesaponins should have the following maximum absorption peaks:max271nm415nm514nm 10g s3.994.063.6 .