IR won't tell you anything useful (with the naked eye, at least). Conjugated pi-bonds absorb in the UV range. Near UV isn't very energetic, and UV in general doesn't penetrate materials very well.

see that light blue light in the lower left of my avatar? that's 253.7 nm UV-C light.. it doesn't even
 penetrate polypropylene very effectively.

Xman that link tells me I have to wait for 184 seconds, BUT, the clock doesnt count down!!!

thanx anyway

F1

Im gona throw my post in here since it is relevant to the topic

Iv been very interested in solid phase extraction of c. paspali fermentation broth.
There are many ways to do this

US patent 4237291
Quote

    EXAMPLE 5

    30 l Culture solution of Clavicepts purpurea IMET PA 134 at its natural pH value (5-6) is stirred for 30 minutes with 1.2 kg bentonite. It is then filtered through a layer of calcium sulfate dihydrate and the filtrate discarded. Drying of the mycella-adsorbent mixture according to Example 1 yields 2.8 kg of the dry mixture, which contains practically all of the alkaloid content of the culture suspension (ergosine, ergosinine, traces of chanoclavine). Rotary extraction of the alkaloids with a 10 to 12 fold volume of ethyl acetate yields up to 90% of the alkaloid content of the dry mycella-bentonite mixture.


This method works with most montmorillonite compoud.  Calcium bentonite is used because its easier to filter and doesn't make a colloid like Sodium bentonite.  Fullers earth I think will work better because it does not form clay as easily, and comes in granules, even as cat litter.  Also it works without any type of acid activate, apparently that helps with the bentonite.  The drawback with the bentonite method is the fact that you have to dry it or extract from it, which is messy and slow to filter.  Fullers earth in a flow-through adsorption column would probably work better.  These types of materials adsorp best at the natural pH of the claviceps broth.  Eluting is done with methanol, or water with base or just a mixture.  Eluting with methanol and or base is more efficient.  I think eluting with base is more prone to cause epimerization, especially with methanol involved tho.

Czech Patent 269105
This uses c18 silica
Quote

    Partial Translation:
    1L filtered fermentation liquid containing [peptide alkaloids] was adjusted with aqueous ammonia solution at pH 6.8 This solution was added 35 g of silica gel 2 x and the bound oktadecylovou velikosticastic? group of 50um (Separon SGX C18) and mixed 15 min on a magnetic stirrer. The mixture was filtered, washed filter cake 100 ml of distilled water and drained. Filter cake was extracted 2 x 150 ml of methanol containing 5% aqueous ammonia. Extracts were combined and concentrated in evaporators. 70% yield


This is very interesting because silica gel is cheap, and is a good flow-through method unlike the clay methods.  Scaled up of course there will be less solvent required.I believe this method is not optimized for c. paspali which contains mostly water soluble ergoamides, whereas c. purpurea methods will contain mostly water insoluble ergopeptides and methods will differ, especially type of optimal solid phase adsorbant used.


Japanese Patent 2-167051
Quote

    Full translation
    Specification

    1. Name of the invention
         Separation and purification methods of ergot Alkaloid

    2. Scope of Patent Request
       1.  Separation and purification methods of ergot Alkaloid which has a characteristic like dissolving the Alkaloid by Dissolving agent after adsorption of ergot Alkaloid by having Fermentation liquor possessing ergot Alkaloid that can be obtained by culturing Ergot Claviceps paspali contacting a kind of Synthetic adsorbent porous polymer that can be selected from Styrene-Divinylbenzene Copolymer and Phenolic resin.

    3. Detailed explanation on the invention
       
    Application area in the industry

    This invention is regarding how to achieve separation and purification of ergot Alkaloid out of Fermentation liquor that can be obtained by culturing Ergot Claviceps paspali.
    Ergot Alkaloid is a useful compound known for a variety of efficacy as a medical product.
       
    Traditional Technology

    Fermentation liquor of Ergot Claviceps paspali is an Aqueous solution that mainly contains 8:2 mixture of ?-Hydroxyethyl lysergic acid amide and lysergic acid amide as well as sugar which is culture component, ammonium salts of organic acids, metal ions and a variety of foreign substances such protein, fat, pigment, which are the components of metabolite and body fungus.
    Traditionally, the standard method of separating ergot Alkaloid was to extract it by organic medium such as Chloroform, Ethyl acetate, and n-Butanol. However, in order to improve the collection rate of ergot Alkaloid, this method requires a lot of solvent. Another problem is that it was hard to avoid emulsification caused by protein, fat, and defoamer contained in fermentation liquor.
    The other method that uses resin is explained in Patent-Application-Sho-45-12303 to use strongly acidic cation ion exchange resin. However, for ion exchange resin, a special processing is required for the resin to be activated or regeneration. Besides, there are many challenges for the practical application such as swelling and heating. Patent-Application-Sho-54-90198 has Separation and purification methods of ergot Alkaloid using insoluble polymer containing substituent like Alcoholic hydroxyl group. However, the content is about the method to separate mixture of peptide type ergot Alkaloid such as Ergocristine, Ergocryptine, and Ergocornine.

    Challenge to be overcome by this invention

    As mentioned above, up until this point, there is no practically satisfactory method established in the industry for separation and purification of ergot Alkaloid out of fermentation liquor.
    This invention provides a new method of separation and purification of ergot Alkaloid.

    Method to resolve the challenge

    This invention is regarding how to achieve separation and purification of ergot Alkaloid out of Fermentation liquor that can be obtained by culturing Ergot Claviceps paspali.

    Separation and purification methods of ergot Alkaloid which has a characteristic like dissolving the Alkaloid by Dissolving agent after adsorption of ergot Alkaloid by having Fermentation liquor possessing ergot Alkaloid that can be obtained by culturing Ergot Claviceps paspali contacting a kind of Synthetic adsorbent porous polymer that can be selected from Styrene-Divinylbenzene Copolymer and Phenolic resin. Fermentation liquor produced by Claviceps paspali has full of amide type ergot Alkaloid such as ?-Hydroxyethyl lysergic acid amide and lysergic acid amide. Even though it is also ergot fungus, the fermentation liquor of Claviceps purupurea mainly consists of peptido type ergot Alkaloid, these chemical structure as well as physical and chemical characteristics are different.
    For synthetic polymer absorbent used for this invention can be Divinylbenzene-styrene Copolymer and Phenolic resin. These synthetic polymer absorbent does not have remarkable functional group in the molecule, but simply, absorbs the compound by Van der Waals force possessed by the molecule. Generally speaking, we use the ones with grain diameter 250~857 um, pore diameter about 500 A, and the surface space is 400 ~ 1000 m2/g. For example, styrene-divinylbenzene Copolymer, Sepabeads SP207, SP206, SP800, SP900 by Mistubishi Chemical, Duolite S-866 and S-861 by Rohm and Haas. For Phenolic resin, duolite S-761.

    Though Synthetic polymer absorbent is able to process fermentation liquor by batch, the method like passing fermentation liquor by getting packed in a column is easier and better. The amount of resin to be used depends on the absorbent power of ergot Alkaloid 60 mg/ml - wet resin. The size of column is not specified, but the column length is better to be 2 to 5 times longer than the inner diameter of the tube.

    Because of Hydrophilicity of amid type ergot Alkaloid, organic eluent that contains 5~80 weight % of water can be used for this invention. As organic eluent, water soluble alcohol type like methanol, ethanol, n-propanol, secondary propyl alcohol, and water soluble ketons like acetone, water solule ether like tetrahydrofuran and dioxane, and water soluble nitrile like acetonitrile are used, and can be used either as single solvent or compound solvent of those. Also, you may add alkalescent component such as Sodium hydrogen carbonate aqueous ammonia or weakly acidic component such as succinic acid and citric acid into the solvent system.

    Next, example steps performed for this invention will be explained.

    After filtered from fungus body, fermentation liquor of Claviceps paspali can be directly used for this experiment method. However, because the pH of the liquor is 5.0 ~ ., it is better to add ammonia water to increase pH to over 7.0 if you wish to improve yield of ergot Alkaloid. This way, it is possible to collect ergot Alkaloid constantly. Pack synthetic polymer absorbent into the column in advance, then, wash it by water. The passing speed of fermentation liquor to go through the column packed with synthetic polymer absorbent should be 0.5 ~ 2.0h. After fermentation liquor has been passed through, next, pack eluent into the column. the amount to be used and the speed of the eluent depends on the type of the eluent, but in general, 3 to 6 times larger than the content of the resin. You may decide whether or not the elution of the Alkaloid has been completed or not by thin-Layer Chromatography. It is easy to extract ergot Alkaloid with high purity by removing the remaining liquor including water using extracting solvent like ethyl acetate after removing elution under the decreased pressure. The used Synthetic polymer absorbent can be easily reused after being washed by secondary propyl alcohol including 1N sodium hydroxide aqueous solution or acetone, followed by water wash. Below, more details of this invention with application examples:

    Example

    Example1

    After Fermentation liquor of Ergot Claviceps paspali 500 ml (including ergot Alkaloid 1040mg, pH5.2) went through a column packed with 20ml of styrene-divinylbenzene Copolymer, Sepabeads SP207 (Mitsubishi) by speed 5ml/min, wash it with 80ml of water, then, eluted with 50% methanol water, 80ml. Under the decreased pressure, the methanol is removed from the elution, and adjusted it to be pH9 by adding ammonia to the remaining liquid, extract by ethyl acetate, then, remove ethyl acetate, then let it dry, resulted ergot Alkaloid with 92% purity, 768mg (yield 68%).

    Example2

    Using ammonia water, the fermentation liquor used by Example1, 500 ml (including ergot Alkaloid 1248mg) was adjusted to pH9. Then, let it go through the column with 20ml of Sepabeads SP207 (o20x64mm) with the speed 5ml/min. This gets washed with 80ml of water, followed by eluting with 85% secondary propyl alcohol 80ml. Same process as used in example1, esulted ergot Alkaloid with 94% purity, 1314mg (yield 99%).

    Example3

    Using ammonia water, the same fermentation liquor used by Exampe1, 1000ml (including ergot Alkaloid 832mg), was adjusted to pH9. Then, let it go through the column packed with Duolite S-861 (Rohm and Haas) 20ml (o20x64mm) with speed 5ml/min. After washed by 80ml of water, eluted with 50% acetone solution 80ml. Same process as in Example1, resulted ergot Alkaloid with 90% purity, 915mg (yield 99%).

    Example4

    Using ammonia water, the same fermentation liquor used by Exampe1, 1000ml (including ergot Alkaloid 832mg), was adjusted to pH9. Then, let it go through the column packed with Duolite S-761, 20ml (o20x64mm) with speed 5ml/min. After washed by 80ml of water, eluted with 50% acetone solution 80ml. Same process as in Example1, resulted ergot Alkaloid with 92% purity, 859mg (yield 95%).

    Effect of the invention

    This invention enabled us to easily collect high purity ergot Alkaloid with high yield out of fermentation liquid containing ergot Alkaloid.


Intersting note that the resin they use in the first few examples has a trade name in the US of XAD-4.  This patent is optimized to the alkaloids in C. paspali and is as good as it gets.  I think eluting with mostly methanol would be awesome because it would be easy to remove, and base is not required with this polymer extraction.  Ph of the broth needs to be adjusted more basic to optimize extraction, I have some references which explain why this is so, will attach later on.  It may be possible elute with tartaric methanol/h2o which would stabilize the alkaloids as they come out to prevent epimerization.  This other xad-4 patent says lowering ph is not optimal for this but they refer to other alkaloids.  As usual it might help elute with somewhat basic solution but may not be necessary.

--
If you don't want to work this all up with a/b and solvents after evaping the methanol I see two options:

1)
Elute using mostly methanol and then add excess tartaric acid, remove most solvent then add 10x ether or maybe hexane to precip the tartaric salt of LSA's.  All this work will probably convert LSH to LSA by the time the tartaric hits it.  I assume the 5% water which must be present for elution may disrupt crystallization using this method?

2)
Elute using mostly methanol directly into a nitrogen purged boiling KOH solution with a distillation head on it.  Most of the methanol should boil off as it is eluted during the 2hr+ hydrolysis.  Once all methanol is added and hydrolysis no longer off gasses ammonia then lower temperature add sulfuric acid to low pH

h2so4 is used because the hydrogen sulfate is much less water soluble than the hcl for crashing out. 

I think DVB-styrenic with carboxylic acid (WCX), or sulfonic (SCX) functional groups would be the ticket.
(ex. Dowex-50X8, Amberlite IR120 H+/Na)
polars (sugars) don't adhere to the resin, and are washed out with water, and the alks are eluted according to increasing pKa with methanolic ammonia.

alternately, non-ionic resins may be employed, relying on primarily van der waals interactions
(ex. Amberlite XAD-7)

also a note to you overunity alkaloids get epimerized usually by locally high ph say if you add base and don't stir well.
ph 8-9 won't cause a lot of inversion like you suggest, i don't believe.
trying to elute a cloumn with tartaric acid solutions is a non starter if the acidity of the resin has a higher acidity i think it will stay bound to the resin there so something else to consider.
but since were talking about sepabeads your preference citric tartaric acid would be fine.
and we all know it's most stable as a conjugate acid base pair.
maleic acid is the acid of choice to prevent oxidation to lysergamides as the acid is an antioxidant in it's own right.
that's your best bet.

damn your goood
so your saying purify and seperate epimers in one shot that's dandy please go on.
and why complex with g-cyclodextrin? i wonder

aha,  it was a CE application:

A mixture of ergometrine, ergotamine and
their epimers was separated by capillary zone
electrophoresis in 0.1 M phosphate buffer (pH
2.5) with detection at 206 nm (79). Addition of
~,-cyclodextrin (30 mM) to the background
electrolyte increased both the migration time
and the resolution of the alkaloids. Separation
of five natural ergopeptine alkaloids (ergo-
metrine not included) and their epimers could
not be achieved without both 20 mM [3-cyclo-
dextrin and 8 mM ,{-cyclodextrin in the 50 mM
phosphate buffer (pH 2.5); detection was by
UV at 214 nm or laser induced fluorescence
with an excitation wavelength of 325 nm (80).
Capillary zone electrophoresis was applied to
the analysis of rye ergot sclerotia but not yet to
grain foods.

doesn't matter that this is an electrophoretic analytical method. this may be employed on the
preparative scale using hydrodynamic flow

thanks

chromatography has been an interest of study and practice for the past three years, in school and industry.
ergolines have been an interest for the past 18 years (heh heh).
I have some Amberlite 252 Na on the way, and an ace column attached to a vac filtration flask; will
do some experiments based on the patent overunity33  graciously posted

I'll address epimer separation later, using microflowLC-UV

i missed this methyl ester business...what is it about?
I'll tackle the specifics of a stationary phase, but am open to elution ideas