Derivatization is one way, but more laborious and expensive. You can try the more straightforward approach of indirect UV detection.  The detection of a UV-Vis-transparent analyte is accomplished by adding light-absorbing species into the mobile phase. The presence of the analyte is monitored by measuring a decrease

in the light absorbed by the eluent as the analytes elute from the column. To select the right visualization agent the following publication by Xiaoxuan Shen and Sterling A. Tomellini will be very helpful:

https://www.google.gr/url?sa=t&rct=j&q=&esrc=s&source=web&cd=9&ved=0ahUKEwjZxYbpkZHWAhWCWBQKHYmaCCIQFghjMAg&url=http%3A%2F%2Fquimica.udea.edu.co%2F~carlopez%2Fcromatohplc%2Freview_Indirect_Photometric_Fluorometric_Detection_hplc_2007.pdf&usg=AFQjCNE4sDNgm6VCvlRUOd3DZmzB-ogHOQ

Good luck.  

The only way you could analyze a UV inactive compound on a UV detector is to derivatize it to add a UV active functionality, such as a phenyl ring.  Otherwise you can't use UV on the compounds in their native state since there is nothing for the UV detector to detect. If you are doing this as a preparative LC than derivatization wouldn't be an option.  Your only recourse would be to use another type of detector.

Your only chose to make this molecule UV absorbent is to tag (derivatize) it.  However, be sure you don't derivatize the chiral center(s).  Cis-trans isomers will open, then rotate their bonds before closing again.

 Not everything has a chromaphore and not everything can be seen by UV/VIS. Just use an alternative type of detector with your HPLC (e.g. Polarimeter, RID, MS, ELSD, CAD...). *Make sure you confirm that your sample is achirally pure BEFORE you attempt to develop an HPLC chiral method of separation!

Derivatization is one way, but more laborious and expensive. You can try the more straightforward approach of indirect UV detection.  The detection of a UV-Vis-transparent analyte is accomplished by adding light-absorbing species into the mobile phase. The presence of the analyte is monitored by measuring a decrease

in the light absorbed by the eluent as the analytes elute from the column. To select the right visualization agent the following publication by Xiaoxuan Shen and Sterling A. Tomellini will be very helpful:

https://www.google.gr/url?sa=t&rct=j&q=&esrc=s&source=web&cd=9&ved=0ahUKEwjZxYbpkZHWAhWCWBQKHYmaCCIQFghjMAg&url=http%3A%2F%2Fquimica.udea.edu.co%2F~carlopez%2Fcromatohplc%2Freview_Indirect_Photometric_Fluorometric_Detection_hplc_2007.pdf&usg=AFQjCNE4sDNgm6VCvlRUOd3DZmzB-ogHOQ

Good luck.  

We did using ELSD or RID detector. Nice if you can use LC MS or LC MS MS

I noticed you said you want to separate and detect chiral compounds. A simple derivatization method might not work because it might cause racemization. You don't want to add anything in the mobile phase that might decrease your enantioresolution. I did my dissertation on chiral separation, I might be of help if I know the exact compound you're trying to separate and the equipment which is at your disposal. Without that info - I suggest using a chiral derivatization reagent. 

Hi Kirtansinh

The use of the indirect absorbance method would be the best answer to your problem.

As you are performing a chiral separation it is not a good idea to add a large concentration of absorbent background compound to the mobile phase used for the chiral separation that could alter the separation. Adding the background compound post column through a T-junction allows you to adjust the background absorbance to suit the concentration of your analyte as the sensitivity of the method will depend on the final absorbance of the background mobile phase. The flow rate will also need to be slow and optimised as turbulence at the addition point could mix the background additive with the analyte making the addition useless.

Good luck.

I suggest to complex the compound with some kind of regents to give a color so you can detect in visible range. For the enantiomer you can modify a mobile phase containing 5% beta-cyclodextrine to enhance selectivity towards enentiomers.

It is an important question. Sugars and Cholesterols are surely difficult to determine by the photometric detection. These molecules are determined by RID or Refractive Index Detection, but not so sensitive.

Derivatization of sugars may be achievable by using Pyridylamination (PA) method, which uses a machine. However, I do not sadly find a good derivatization method for Cholesterol yet.

I have used 1,2-diamino-4,5-methylenedioxybenzene dihydrochloride (DMB) to determine N-acetyl-neuraminic acid (NANA)/sialic acid (please see file; NANA determin).

I also have used 9-anthryldiazomethane (ADAM) to determine long-chain fatty acids, biotin (vitamin H), and lipoic acid (thioctic acid) (please see files; Netherlands Biotin and Lipoic acid Avidin).

Amino acids have been derivatized by phenyl isothiocyanate (PITC; aminoacid sequencing grade) (picoTag method of Waters Co., or please see files; PTH Roma RP-HPLC and D-Asp Avidin).