I have several decades of experience in finding simple, efficient and accurate chiral GC, CE, HPLC and SFC methods.

Method development for chiral samples is different than it is for achiral samples. Column choice is the most important factor. Get the column right and the method is not difficult. Unlike conventional RP/NP separations where one or two popular columns can often provide some initial results, chiral columns do not work this way. Here are some general info and links.

Sample solubility is extremely important so it should be your first consideration as this will suggest which mode of separation is more appropriate (NP/RP). Also, will the method need to be scaled up to collect samples? If so, NP is often preferred.

Next, check the available literature for any examples. A simple keyword search on GOOGLE may provide some examples. If you find published chiral methods, that should not mean you use them w/o more thought (a good percentage of the published chiral methods are of very poor quality. Check the sample's K prime and peak sym). Use all of the "free" information you find to develop a good quality chiral method. Finding the best column for the sample is always the first step. BTW: Some column manufacturer's will screen THEIR columns for free or for a fee to see if they have something that will work for you (This can be a very economical way to find out which column(s) show promise w/o purchasing them all).

I have linked several free articles on CHIRAL METHOD DEVELOPMENT below to help you. Please review them for more information.

"Chiral HPLC and SFC Column Screening Strategies for Method Development: "

"CHIRAL HPLC / SFC METHOD DEVELOPMENT (Alcohols):"

"Some Stereochemistry Terms (Chirality)"

http://hplctips.blogspot.com/2011/06/chiral-hplc-and-sfc-column-screening.html

http://hplctips.blogspot.com/2011/04/chiral-hplc-sfc-method-development.html

http://hplctips.blogspot.com/2011/06/some-stereochemistry-terms-chirality.html

I have several decades of experience in finding simple, efficient and accurate chiral GC, CE, HPLC and SFC methods.

Method development for chiral samples is different than it is for achiral samples. Column choice is the most important factor. Get the column right and the method is not difficult. Unlike conventional RP/NP separations where one or two popular columns can often provide some initial results, chiral columns do not work this way. Here are some general info and links.

Sample solubility is extremely important so it should be your first consideration as this will suggest which mode of separation is more appropriate (NP/RP). Also, will the method need to be scaled up to collect samples? If so, NP is often preferred.

Next, check the available literature for any examples. A simple keyword search on GOOGLE may provide some examples. If you find published chiral methods, that should not mean you use them w/o more thought (a good percentage of the published chiral methods are of very poor quality. Check the sample's K prime and peak sym). Use all of the "free" information you find to develop a good quality chiral method. Finding the best column for the sample is always the first step. BTW: Some column manufacturer's will screen THEIR columns for free or for a fee to see if they have something that will work for you (This can be a very economical way to find out which column(s) show promise w/o purchasing them all).

I have linked several free articles on CHIRAL METHOD DEVELOPMENT below to help you. Please review them for more information.

"Chiral HPLC and SFC Column Screening Strategies for Method Development: "

"CHIRAL HPLC / SFC METHOD DEVELOPMENT (Alcohols):"

"Some Stereochemistry Terms (Chirality)"

http://hplctips.blogspot.com/2011/06/chiral-hplc-and-sfc-column-screening.html

http://hplctips.blogspot.com/2011/04/chiral-hplc-sfc-method-development.html

http://hplctips.blogspot.com/2011/06/some-stereochemistry-terms-chirality.html

For chiral columns, it requires far more complex "guessing" as compared to RPLC or GC. A chiral mixture is separated by three-point interaction in a chiral environment provided by the stationary phase chemistry. One should be aware of the class of compound you are trying to separate. For example, is it a primary anime? Is it an acid? etc. By reading the column literature one can narrow down the choices of columns by the classes of compounds they separate. Secondly, there are broad selectivity columns such as amylose/ cellulose bonded silica, which will separate large classes of compounds. You can read several online booklets by Sigma on chiral separations

In the pharmaceutical screening process, where there is no shortage of money, columns, and automatic instruments, brute force approach is used where several columns can be tested automatically to get good hits (using special plumbing and valves) with several mobile phases. Promising columns are then chosen and then optimized.

Hello Bill...I would like to know what is meant by RP/NP in chiral HPLC.

So far samples solubility is considered, all my samples are soluble in isopropanol so how does that provide me any leads in choosing a column or the polarity for a new compound?

Can you please provide me some examples of keywords which I can use in Google to narrow down my range of choices either for a specific new chiral compound or for a particular class of chiral compounds.

Some previously established methods based on published results are available from supporting info of journals accessible via Scifinder.

I use HPLC of Agilent Technologies.

Arnab: It sounds like you may be a new student to HPLC, so before starting out on a complex project like this, please get some local help from someone with several years of practical chromatography experience to help you. Read everything you can find on the topic and get lots of help. You will learn much faster this way. To become proficient at liquid chromatography takes many years and thousands of samples to learn on. Many types of chiral chromatography methods can be even harder to learn, especially without a clear understanding of the fundamental terms, calculations and methods used. Did I mention that getting the help of an experienced local person(s) is the key to your success? (That is the best advice).

Note: We are talking about developing ONE method in this discussion thread. The links that I supplied discussed methods and techniques used to develop a chiral HPLC or SFC method. Each sample will have its own issues and solutions to find an HPLC method. "Classes" of similar compounds often will require different columns and methods, not one. Some exceptions exist. *More details would be needed to know the answer and we are generalizing at this point.

OK, "RP" means Reversed Phase and "NP" means Normal Phase.

If your compounds are well solubilized in "IPA" (Isopropanol), then you may want to consider NP methods and columns. Perhaps a broad spectrum sugar column would be a good place to start (cellulose or amylose chiral phase). Success with chiral method development requires that you FIRST develop an achiral (not chiral) HPLC method to determine the sample's chemical purity. Please do this before you make any attempts to resolve your racemate using a chiral method.

Keywords for Google or Scifinder: As examples, I would suggest you use the chemical name and/or chemical class of  your sample. Be specific. The word "chiral" would also be a good choice of keywords. Words such as "HPLC", "Method" or "Separation" may also be useful. *SciFinder will only lead you to journal abstracts, but Google will lead you to many more resources as just about "everything" is on the web these days.

I would like to recommend the HPLC-Avidin-affinity column.   This affinity column or gels (Bioptic AV-1,  5μm diameter silica gel) is obtainable from GL Sciences Inc., Tokyo, Japan.   This conclusion has been derived from my experience to separate D-Asp from L-Asp (please see file; D-Asp avidin), since the other commercial HPLC-chiral- separation-columns cannot reliably or reproducibly separate them.

Dear Arnab, here is the link for the development of chiral method on basis of structural component of your compound. Here you will be get mobile phase and column to be used for your best separation.

https://search.daicelchiral.com/name

AS you mentioned above you are using HPLC of Agilent technologies. For separation of your compound you can use the column CHIRALPAK  AD-H(Particle size 5μm,Dimensions 4.6mmIDx150ml, Lot NO. ADHOCD-OF016).You can use the method, n-Hexane / 2-propanol solvent mixture (98:2 v/v) as mobile phase with flow rate 0.2ml/min.

 It`s decided by experience. Before enantioseparation, structure of molecule also plays an important role in column selection.