New Retention Possibilities for Polar Compounds using Cogent Diol Column

TYPE-C Silica™ columns can be used for separation of a variety of polar compounds.  The new Cogent Diol™ column makes an excellent addition to these stationary phases. Consisting of a short alkyl chain with vicinal alcohol groups, this material has additional separation capabilities compared to other phases. Since it is on a TYPE-C Silica™ surface, the bonding is very strong and durable. The diol group can interact with analytes via hydrogen-bonding interactions, resulting in new selectivity. 

In the following example, we demonstrate the capabilities of this intriguing material. The test solutes chosen for the separation were seven common hydrophilic vitamins. The results show that baseline separation could be obtained for all seven analytes. The retention range is adequate as well, with no analytes eluting at the solvent front or sticking to the column. Peak shapes were sharp and symmetrical, and no tailing was observed. 

This column makes a wonderful addition to the TYPE-C Silica™ line of HPLC columns and should be a great benefit to every chromatographer’s toolbox. 

1. Ascorbic acid
2. Niacin
3. Riboflavin
4. Folic acid
5. Pyridoxine
6. Metformin
7. Thiamine

See the following link for full details:

http://kb.mtc-usa.com/article/AA-02102/0/ 

Simple Strategies for Improving Analyte Recovery

Suppose you’re doing an HPLC assay of a basic pharmaceutical and find a 66% recovery for the API. What happened? Did the drug not extract completely? Did degradation occur? These types of scenarios take up valuable laboratory time and resources with investigations, troubleshooting, and re-run analyses. In this case, it may be simply due to the autosampler glass. Regular glass has silanols on the surface that can bind with basic analytes and therefore lead to lower recovery. However, this problem can be prevented with some simple considerations.

In this example using a basic test analyte (cetylpyiridinium chloride), we can see how use of either an ammonium acetate buffer (red trace) or a formic acid additive (blue trace) can reduce the effect of analyte loss compared to DI water alone (green trace).  Using 0.1% formic acid, the silanols become protonated and neutral, which prevents ionic interactions with the analyte from occurring. This is the more effective of the two additives. Ammonium acetate helps the problem by a different approach. The ammonium ion competes with the analyte for the silanols groups, and so fewer sites are available for analyte loss to occur.

Here we use a different strategy for reducing the analyte loss. Reduced Surface Activity (RSA™) glass is made with almost no surface silanols and we can see a major improvement for analyte recovery compared to regular glass. For best results, you can use a combination of RSA glass vials and a formic acid diluent. For solubility reasons though, sometimes you might have to use an ammonium acetate diluent. In that case, RSA glass shows significantly better recovery than regular glass.

                When analysts plan a sample preparation procedure, they may put much thought into the extraction method, dilution procedures, and so on. However, the role of the vial is often overlooked.  Vials are inexpensive and disposable, so they can be seen as relatively unimportant in the analysis process. This data shows how they are not inert and can significantly skew your analytical results. A careful selection of vial and diluent is all it takes to prevent your laboratory from experiencing these kinds of problems.

Eastern Analytical Symposium is Coming Soon!

This year at EAS 2013, MicroSolv will have a booth showing new products and technology at location # 611. In addition, we will be presenting two technical posters in the “Applications of HPLC and UHPLC” segment on Wednesday November 20, 2013 from 12pm–2pm.  We will be exhibiting these posters at booths 418 and 419. This is an excellent opportunity to discover what products we offer that can benefit your laboratory as well as the real world applications using these products, as demonstrated with the poster sessions.

                Among the most notable of our latest products is the 2.o™ line of TYPE-C Silica columns. These items are near-UHPLC stationary phases (2.2µm particles) that will help to save your lab valuable analysis time with increased throughput. All the benefits of our innovative TYPE-C Silica™ are combined with high-efficiency small particle capabilities to allow for great potential for a variety of fields such as metabolomics and clinical analyses. Our 2.o™ line of products has shown great lot-to-lot reproducibility and durability in the field. Stop by our booth and learn more about this exciting product and how it can help your laboratory become more productive.

                For more information on the symposium, be sure to check out the EAS homepage.

Visit our Poster Sessions at Eastern Analytical Symposium

This year MicroSolv will be presenting two technical posters at Eastern Analytical Symposium in Somerset, NJ. The presentations will be held in the “Applications of HPLC and UHPLC” segment on Wednesday November 20, 2013 from 12pm–2pm.  Please stop by our booths and learn how your laboratory can benefit from these exciting new technologies.

                The first presentation is entitled “Advantages of Reduced Surface Activity (RSA) Glass Autosampler Vials for Basic Solutes.” In this study, our acclaimed RSA™ autosampler vials were compared with those of a market leading manufacturer based on conventional borosilicate glass. A variety of test solutes were assayed at different time intervals and concentrations using both vial types. The results of the study led to a number of interesting conclusions. First, only solutes containing basic functional groups showed any loss in peak area after a given time interval had elapsed, suggesting that interaction with silanols on the glass surface was responsible for the change. Second, the peak area loss was found to be drastically lower when using the RSA vials vs. the ordinary vials (see Fig. 1), which is believed to be due to the mostly silanol-free surface of the former. Third, the effect was more significant at early time intervals and leveled off after about 4 hours. This effect could lead to problems with precision as well as accuracy if different peak areas were obtained for consecutive injections in an analysis.

                In the second presentation “Separation of 1,3-Dimethylamylamine and Other Polar Compounds in Dietary Supplement Formulation Using Aqueous Normal Phase Chromatography with Mass  Spectrometry,” we use the Diamond Hydride™ HPLC column to separate hard to retain analytes. In a workout supplement called Jack3d®, each polar compound is retained and separated without the use of ion pair agents, making the method suitable for LC-MS. One ingredient (1,3-Dimethylamylamine) is controversial in terms of safety for consumption and has already been banned in a number of countries. As more data is accumulated for demonstrating the potential hazard of this compound, a need will arise for accurate and reliable quantitation in a variety of matrices. Therefore the Diamond Hydride™ column can play a significant role in these analyses.

                A technical program of the poster sessions is available here. We look forward to seeing you at the symposium this year!

 

Figure 1. Percent recovery of 5.00 ppm cetylpyridinium chloride solution at time intervals of 1 hour each for 4 hours. Both vial types are compared.

How is the amber color produced in the polypropylene/glass amber autosampler vials?

The amber color present in some of the autosampler vials is created by organic or inorganic pigments or dyes. In glass vials, the amber color is from iron oxide, added in less than 1% concentration.
       In plastic (polypropylene) vials, the color is produced from a proprietary organic compound present in mostly less than 0.1% concentration so no iron is used making it appropriate for Ion Chromatography. At concentrations greater than ~2%, the vial becomes completely opaque. The full list of items which have an amber color is given as follows:

Catalog Number	Material
9502S-PP-A	plastic
97040-0AV	glass
9502S-WAV	glass
97060-0AV	glass
95002-0AP-A	glass + PP
95010-0AV	glass
95020-0AV	glass
95025-CT-10A	glass
95025-CT-20A	glass
95025-PE-20A	glass
9502C-0-WAV	glass
9502C-WAV	glass
9503S-WAV	glass
9504S-0AV	glass
9504S-WAV	glass
9532C-TS-A	TPX
9532S-TS-A	TPX
97015-AV-08	glass
97015-AV-12	glass
97018-SP	PP + Butyl/PTFE
97020-0AV	glass
97030-0AV	glass

     The purpose of the amber color is for cases in which light sensitive compounds are involved. Folic acid for example is sensitive to photo-oxidation and therefore the effects of ambient light could be detrimental to accurate analyte quantitation.
     There are two types of chemicals which are introduced to a substrate to produce color: pigments and dyes. Although the two terms are often used interchangeably in everyday language, they are actually distinct. Pigments are insoluble in the substrate in which they are infused and are present as a dispersed suspension in the material. Dyes on the other hand are soluble in the substrate and are present as a solution. In the amber products discussed above, the iron oxide in glass is a pigment and the proprietary organic compound in polypropylene is a dye.
      A wide variety of colors besides amber can be produced in glass depending on the colorizing agent used. The green color used in wine bottles (Fig. 1) for example is produced by a combination of iron oxide and chromium (in the form of chromic oxide or potassium dichromate). Copper produces a turquoise color notable for use in Egyptian Blue, a widely used synthetic pigment in antiquity (Fig. 2). Cobalt gives a deep blue color which has been found in many examples of ancient Chinese porcelain (Fig. 3). Also, glass coloring is not necessarily produced by the addition of pigments or dyes. Due to the Tyndall effect for example, color in glass can be achieved by light scattering in a suitable medium.

Fig. 1. Coloration in wine bottle

 

Fig. 2. Example of Egyptian Blue in an ancient artifact

 

Fig. 3. Cobalt used in Chinese porcelain.

Clear Polypropylene 2ml Vial Information
Amber Polypropylene 2ml Vial Information