07-01-02 04:27
No 327450
I was walking through a field one day with a friend and he spotted some ergot growing on rye. We picked it and decided to grow it in a agar water mixture. Its growing great but i Need to know how to extract the alkaloids using OTC chemicals. I also don't have access to tartaric acid or chloroform.
(Hive Addict)
07-01-02 06:58
No 327509
../rhodium /chlorof
Post 272739 (terbium: "Re: Cream of tartar for D-tartaric acid", Chemicals & Equipment)
(Hive Addict)
07-01-02 23:17
No 327700
Although it's possible, it's extremely expensive. Btw, the calculations should really read 6.4 gallons of 10.8% bleach per 340g of acetone.
Why not use methylene chloride? It has all the solvent properties you desire. Good luck with the procedure btw.
(Hive Bee)
07-02-02 00:25
No 327719
that was a dream you had right? ok. lycaeum has a good doc on ergot alkaloid extraction. link at bottom of page.
(Hive Bee)
07-12-02 13:14
No 331672
The proper way to do it:
(Sorry about huge fonts but at least it's clear
)
(Hive Bee)
07-12-02 13:16
No 331673
(Can't remember the source..)
(Hive Addict)
07-28-02 23:25
No 338166
Heres the above post by mountain girl in readable text.
Indole alkaloids.
All the members of this group of alkaloids contain an indole nucleus in their chemical structure; however, they are derived from many different natural sources. Indole alkaloids are the principal alkaloids isolated from ergot, Strychnos nux vomica, and rauwolfia. Each of these groups will be treated separately.
Ergot alkaloids. Ergot is the dried sclerotium of the parasitic fungus Claviceps purpurea prepared and developed on plants of rye. Ergot yields not less than 0.15% of the total alkaloids of ergot calculated as ergotoxine, and water soluble alkaloids equivalent to not less than 0.01% of ergonovine. Six diastereoisomeric pairs of alkaloids have been isolated from ergot. The most physiologically active alkaloids are of the levo form. All these alkaloids can be considered to be derivatives of lysergic acid or its stereoisomer, isolysergic acid. The principal alkaloids of ergot are lysergic acid (11), ergometrine, or ergonovine (12), and ergotamine (13).
Analysis of indole alkaloids.
Ergot alkaloids. Alexander and Banes have developed a procedure for the analysis of ergot in which the total alkaloids are extracted with ammonium hydroxide methanol chloroform and the water soluble alkaloids are seperated by column chomatography. In this procedure the total alkaloid and water soluble ergot alkaloid contents are determined colorimetrically. The chromogenic reagent, p-dimethylaminobenzaldehyde, first proposed by Smith is used in this procedure. Certain amines condense with p-dimethylaminobenzaldehyde to give products which are oxidizable under strongly acidic conditions to produce a color. In the case of the ergot alkaloids, it is the indole portion of the lysergic acid nucleus which condenses with the color reagent. The detailed procedure of Alexander and Banes follows:
Reagents: The extracting solvent is a solution of 10 ml of concentrated ammonium hydroxide in 90 ml of methanol mixed with 900 ml of chloroform. Prepare the color developing reagent as follows: dissolve 125 mg of p-dimethylamino-benzaldehyde in a cooled mixture of 65 ml of concentrated sulfuric acid and 35 ml of water, and add .05 ml of 10% ferric chloride solution; use within 7 days. To prepare the diatomaceous silica support, boil 150 g of celite 545 with 1000 ml of concentrated hydrochloric acid for 10 min, cool, then wash with water to remove the acid and dry in an oven at 100°C; the washed product should give no acid test when moistened. Also prepare a standard solution containing 20µg of ergonovine/ml as follows: Dissolve 6.78 mg of ergonovine maleate reference standard in 1% tartaric acid solution.
Extraction. Weigh 5 g of ergot and place in a 125-ml sep funnel. Add 50 ml of extracting solvent and shake vigorously for 5 min. Add 50 ml of chloroform and 2 ml of water, shake gently, and allow to settle. Draw off the lower phase as completly as possible, and filter through a glass wool plug into a 200-ml volumetric flask. Add another 20 ml of extracting solvent to the ergot residues in the sep funnel, mix, add 20 ml of chloroform, and shake. Filter the lower phase through the glass wool plug into the volumetric flask. Repeat this procedure once more with an additional 20 ml of extracting solvent and 20 ml of chloroform. Dilute to volume with chloroform.
(Hive Addict)
07-29-02 00:09
No 338187
Prep of total alkaloids assay solution. Evap a 25ml aliquot of the extract solution to dryness at a temp not exceeding 40C. Dissolve the residue in 20 ml of ethyl ether and transfer to a 125 ml sep funnel with the aid of three 20 ml portions of ether followed by a 5-ml and 3-ml portions of 0.2 N sulfuric acid. Shake and draw off the acid layer into a 25ml volumetric flask thru a glass wool plug. Extract the ether layer with three 5-ml portions of 0.2 sulfuric acid. Combine the acid extracts and dilute to 25 mil with 0.2 N sulfuric acid.
Prep of water soluble alkaloid assay solution. Place 5g of diatomaceous silica support and 5ml of 0.1 M citric acid in a beaker. Mix thoroughly and transfer to a 25x200 mm chromatographic tube. Pack firmly with a packing rod. then mix 2g of silica support with 2 ml of water and pack on top of the acid trap. Evap a 150ml aliquot of the chloroform-ammonium hydroxide extract to dryness at a temp not exceeding 40C. Dissolve the residue in 10ml of chloroform and transfer quantitatively to column. Elute with 125ml of water saturated chloroform. Inspect for proper retention of water soluble alkaloids by use of an ultraviolet lamp. A brightly florescent blue ring in the citric acid layer indicates that ergonovine has been properly retained. Discard the eluent. Extrude the column containing the sample with gentle air pressure into a 100ml beaker, and mix with 5ml of 10% sodium bicarbonate solution. Then mix with enough silica support to make a workable mixture. Transfer quantitatively to a chromatographic tube. Pass 75 ml of water saturated chloroform thru the column recieving the eluent in a 125ml sep funnel, containing 5 ml of 0.2 N sulfuric acid. Examine under an ulatraviolet lamp to check for complete removal of water soluble alkaloids from the column.
Shake the sep funnel and draw off the chloroform layer into a second sep funnel. Transfer the acid layer into a 25 ml volumetric flask. Rinse each sep funnel with three 5ml portions of 0.2 N sulfuric acid. Add the acidic rinse to the 25ml volumetric flask and dilute to volume.
Color development. Transfer 5ml aliquots of the two assay solns. and the standard solution to seperate stoppered erlenmeyer flasks. immerse in an ice bath and add 10 ml of color developing reagent to each flask dropwise. Allow to stand at room temp for 45-60 min. Prepare a reference blank by adding 10ml of the color developing reagent to 5ml of 0.2 N sulfuric acid. Determine the absorbance of the assay and standard solutions vs. the blank at 550mµ.
Ergotamine and Ergonovine Pharmaceutical Preperations.
The analysis of tablets and injectable solutions of ergotamine and ergonovine is also based on the use of p-dimethylaminobenzaldehyde as a colorimetric reagent. The detailed procedure for the analysis of ergonovine maleate injectable soln. will be given here. However with only a change in the prep of the sample and standard soln, this same procedure can be applied to analysis of other ergonovine and ergotamine preps.
Procedure:
Prepare the color developing reagent by dissoving 125 mg of p-dimethylaminobenzaldehyde in a cooled mixture of 65ml of concentrated sulfuric acid and 35 ml of water, and add .05 ml of 10% aqueous ferric choride soln; use within 7 days. also prepare a standard soln containing 5mg of ergonovine maleate reference standard 250 ml in water. Accurately transfer a volume of sample solution equivalent to about 2mg of ergonovine maleate to a 100 ml volumetric flask and dilute to volume with water. Transfer exactly 2ml each of standard and sample solutions to suitable test tubes, add to eash 4ml of color developin reagent, mix, and allow to stand in subdued light for 1 hour. Measure the absorbance of the standard and sample solutions at 550 mµ versus a reagent blank with a suitable spectrophotometer.