1. Why should I use CElixir™?



2. Can CElixir™ be used with SDS?



3. Is there any way I can minimize my serum protein samples from aggregating in CElixir™?



4. In your Operating manual for CElixir™, you suggest a water plug. What is the purpose of this step?



5. How do you suggest the water plug be performed?



6. What current should I expect, using CElixir™, at 25kV in a 50um ID Capillary, 27cm long?



7. We have had some trouble getting our sample into solution with CElixir™. What do you suggest?



8. How do I eliminate the band broadening I get on subsequent runs when working with high concentrations of proteins?



9. Will the migration order of my analytes change from my current method when I use CElixir™?



10. Why do you suggest that I use a Water plug with CElixir™?



11. What injection volume should I use when changing my method to CElixir™?



12. Do I use a buffer with CElixir™?



13. Can I use organic solvents with CElixir™?



14. What effect does CElixir™ have on positively charged analytes like Proteins?



15. How do I get started with CElixir™?



16. How do I get a pH other than that of the pH on the bottle of The Accelerator Solution (B)?



17. My method calls for a Citrate Buffer. Will CElixir™ give the exact same results if I run it at similar pH?



18. If I have to develop a new method what is the advantage to using CElixir™?



19. Why does my sample have such less affinity for the wall when I use CElixir™?



20. What is the basic formula of CElixir™?



21. Does CElixir™ require any other reagents to use?



22. How do I store CElixir™?



23. Can I return the capillary to its original state after I use CElixir™?



24. Will MicroSolv Technology Corporation assist me in developing methods using CElixir™?



25. Is CElixir™ Compatible with Urea?



26. What is the 3 Step Method for Separation of biological analytes such as Myoglobin, Carbonic Anhydrase, Interferon-Alpha and HAS?

1. Why should I use CElixir™? [top]
CElixir™ provides you with a reproducible CZE environment. Not only does it stabilize the EOF but also it provides a much reduced wall interaction. There are many other benefits including ease of use with additives for chiral separations.

2. Can CElixir™ be used with SDS? [top]
No, it cannot be used with SDS. If your current method uses SDS but you are not happy with the results of that method, you can choose from other chiral selectors that are compatible.

3. Is there any way I can minimize my serum protein samples from aggregating in CElixir™? [top]
Yes, when separating serum proteins or iso amylase isoCk or iso alkaline phosphotase, it may happen that a protein (immunoglobulin or enzyme) is bound to another (such as an immune complex) or is present under dimer or tetramer form. In such cases you may want to displace the binding by incubation of your samples for 30 minutes at 37C, (100ul of serum) with 1-2 mg of Ficin (Sigma# F3266).

4. In your Operating manual for CElixir™, you suggest a water plug. What is the purpose of this step? [top]
The water plug is to avoid “sample carry over” into the anodic buffer. This water plug also helps to improve resolution, but mostly a water plug improves injection reproducibility.

5. How do you suggest the water plug be performed? [top]
It is easy to make a water plug injection. Firstly, you should only put the water plug after the sample or in other words, then make a post injection water plug. Depending on you capillary size, you should make inject the CEwater for 1 second at 0.5 psi. If the capillary is a 75m, you can inject the CEwater for 10 seconds at 0.1 psi. Be sure to make both your sample and your CEwater injection toward the cathodic buffer and not to “waste” or an empty vial. Use a ramping time of 1 minute.

6. What current should I expect, using CElixir™, at 25kV in a 50um ID Capillary, 27cm long? [top]
The current you should expect at every pH is approximately 80-90uAmps.

7. We have had some trouble getting our sample into solution with CElixir™. What do you suggest? [top]
One of the greatest advantages of CElixir™ is that you can add organic solvents to assist you with solvating your sample and you will not sacrifice EOF. Ethylene Glycol, Methanol and Acetonitrile have been used (up to 25% of the BGE) and none of the benefits have been lost.

8. How do I eliminate the band broadening I get on subsequent runs when working with high concentrations of proteins? [top]
Use a Water Rinse. It is recommended that when your samples are high concentrations of protein or simply a very “sticky” protein, that immediately before you inject the sample, between runs, that you dip your capillary in CEwater. It is further recommended that immediately after the separation, that you dip your capillary in CEwater. This will assist in removing any protein that will stick to the outside of the capillary walls and will be injected as a type of sample carry over. Also, it is imperative that between runs, that you follow the water rinse with injection of .1N NaOH according the CElixir™ Operating and Trouble Shooting Manual.

9. Will the migration order of my analytes change from my current method when I use CElixir™? [top]
It is very possible that the migration order will be different. It is almost certain that the migration times will be different due to the faster EOF. The shapes of your peaks should be sharper as well and you may see acidic compounds that you did not see when using traditional CZE.

10. Why do you suggest that I use a Water plug with CElixir™? [top]
It is generally suggested that all CE injections include a water plug. The purpose of the plug is to increase your precision. The rationale for this is that is does not allow your analytes to diffuse back into “buffer” vial during the injection process; the capillary may become pressure free when it is withdrawn from the vial it is in to change the vials. During this period there can be a back flow of solution. When you capillary is back in the “buffer” vial, before the voltage is reapplied, analytes have been known to diffuse back into the vial. The water plug does not allow this. Thus better precision.

11. What injection volume should I use when changing my method to CElixir™? [top]
It is best to run an Ohm’s Law Plot for you capillary to see the linear range of the usable voltage; injection volume is very dependant upon the temperature and viscosity of your sample. Under most circumstances, it is recommended that you start with the same injection volume that you have used in other CZE methods.

12. Do I use a buffer with CElixir™? [top]
No, there is no need for other buffers while using the CElixir™ HPCE Column Kit. The Accelerator Solution has the “run buffer” built into it as a part of its formula.

13. Can I use organic solvents with CElixir™? [top]
Yes, you can use small amounts of solvents for solubility and efficiency additives. Be careful not to add so much solvent that you dilute CElixir™

14. What effect does CElixir™ have on positively charged analytes like Proteins? [top]
Proteins that are positively charged are ion paired with the Accelerator Solution (B).

15. How do I get started with CElixir™? [top]
It is very easy to use and get started with CElixir™. Simply start with The Initiator Solution (A) and The Accelerator Solution (B) with the pH that is closet to the pK of your analyte. If you do not know the pK of your analyte, simply start with The Accelerator Solution pH 2.5. If you do not get a peak, try The Accelerator Solution pH 9.2. The CElixir™ Operating and Trouble Shooting Manual has very easy to follow instructions for method development.

16. How do I get a pH other than that of the pH on the bottle of The Accelerator Solution (B)? [top]
You can easily get a different pH of The Accelerator Solution (B) by mixing other Accelerator Solutions of higher or lower pH or by adding H3PO4, HCL or NaOH. The CElixir™ Operating and Trouble Shooting Manual has a complete pH Adjustment Chart for your ease of use.

17. My method calls for a Citrate Buffer. Will CElixir™ give the exact same results if I run it at similar pH? [top]
No. CElixir™ creates an entirely different CZE environment and will be very different. Only the end results should be compared to your original method. A new method will have to be used for CElixir™.

18. If I have to develop a new method what is the advantage to using CElixir™? [top]
The advantage is that CElixir™ provides a more reproducible EOF than current CZE methodology. Most CZE methods will change the EOF after several runs through the capillary. Since CElixir™ is a dynamic coating system and it has a greatly reduced wall adsorption equilibria from CZE, the EOF will not change even after 100 runs when operated correctly.

19. Why does my sample have such less affinity for the wall when I use CElixir™? [top]
The answer is very simple, the adsorption equilibria is shifted from your analyte to The Accelerator Solutions. The reasons for this are many but the main reason is the number of charges on your sample is so much lower than that of The Accelerator Solution. The bound Initiator Solution is much more attracted to The Accelerator than your sample and therefore the attraction of the Initiator and the Accelerator not only reduce sample adsorption, it creates a very robust EOF.

20. What is the basic formula of CElixir™? [top]
CElixir™ is a patented, proprietary formula. It can be said that CElixir™ is a system of two polymers that form very strong and stable bonds with the silanol sites of the capillary wall and creates a very strong, pH independent EOF.

Each CElixir-pH™ buffer contains a buffer as well as the dynamic coating already blended to exact specifications. Each buffer contains the following:

Buffer	pH	Buffer Concentration
Initiator:	2.5	Phosphate 75mM
Accelerator pH 2.5	2.5	Phosphate 75mM
Accelerator pH 4.3	4.3	Malic Acid 60mM
Accelerator pH 6.2	6.2	Phosphate 50mM
Accelerator pH 8.2	8.2	Phosphate 40mM
Accelerator pH 9.2	9.2	Borate 150mM

21. Does CElixir™ require any other reagents to use? [top]
No, All of the reagents that are needed to achieve great CZE with CElixir™ are contained in the CElixir HPCE Column Kit: The Initiator Solution, The Accelerator Solution and CE grade water.

22. How do I store CElixir™? [top]
CElixir™ can be stored at room temperature along with other reagents. If you are going to leave the bottle of CElixir™ open for an extended period, you may want to add a little sodium Azide to minimize bacterial growth.

23. Can I return the capillary to its original state after I use CElixir™? [top]
Yes, you can if you follow the procedure in the CElixir™ Operating and Trouble Shooting Manual. However, it is recommended that once you use CElixir™ on a capillary, you dedicate that capillary for the exclusive use of CElixir™. This is due to the lack of effectiveness of the lab technique employed in removing the CElixir™ from the capillary.

24. Will MicroSolv Technology Corporation assist me in developing methods using CElixir™? [top]
Yes, MicroSolv is very interested in helping all scientists develop methods with CElixir™. To get this free service, contact us at our help desk number 1.732.578.1777 or by using this website.

25. Is CElixir™ Compatible with Urea? [top]
Yes, also with Taurine, Betaine and Ethylene Glycol Guanidine up to 10%. This is only a partial list

26. What is the 3 Step Method for Separation of biological analytes such as Myoglobin, Carbonic Anhydrase, Interferon-Alpha and HAS? [top]
Somsen et al reported in the Journal of Chom. A 1083 (2005) 186-192, success using CElixir-pH™ Initiator, Accelerator and a home made run buffer; TRIS/Phosphate pH 8.5 as a three step method. They used the CElixir System (A&B) to coat the capillary but substitute the run buffer with Tris/Phosphate. They selected the buffer based on their samples but the 3 Step method can include using Phosphate, Borate or any buffer that closely matches the Accelerator pH.

Using the 3 Step method has advantages for many biological complex mixtures over conventional CE or even standard CElixir-pH methods when the separation is just not meeting your requirements.