text:
A separation column and a method of making the separation column are provided. The separation column includes a separation channel and a separation medium in the channel. The separation medium includes a porous matrix, and the porous matrix includes a support and a stationary phase. The support includes a metal organic polymer, such as a photopolymer, and the stationary phase includes a bonded phase. The separation medium can be used to separate a sample of analytes.
1. A separation column comprising:
a separation channel; and
a separation medium in the channel, said medium comprising a porous matrix, said porous matrix comprising a support and a stationary phase, said support comprising a metal organic photopolymer from a metal alkoxide precursor, and said stationary phase comprising a bonded phase.
2. The column of
3. The column of
4. The column of
5. The column of
6. The column of
7. The column of
8. The column of
9. The column of
10. The column of
This application is a continuation-in-part of U.S. patent application Ser. No. 09/929,275, entitled "Photopolymerized Sol-Gel Column and Associated Methods," filed Aug. 13, 2001.
The invention relates in general to a separation column, and, in particular, to a bonded phase photopolymerized sol-gel column.
Capillary electrochromatography (CEC) has been regarded as a very promising analytical separation technique that combines the efficiency of capillary zone electrophoresis (CZE) with the selectivity of liquid chromatography. Although CEC has been applied in many different areas, packed-column preparation and low-detection sensitivity remain challenges of this technique. Capillary columns containing small silica packings have been the mainstay of CEC. One disadvantage of packed columns is the fabrication of porous frits of controlled pore sizes, lengths, and high mechanical stabilities. In response to frit fabrication problems, surface-functionalized open-tubular capillary columns and monolithic capillaries are being developed as variants of packed capillary columns. Monolithic capillary columns have received much attention because of the advantages offered in the control of permeability and surface charge.
A major challenge in CEC techniques is the detection of samples containing analytes at low concentration. The lack of sensitivity at low concentration stems from the small sample volume and the short optical path length for on-line detection. Dedicated sample preparation schemes that enrich the target analytes before sample injection are often necessary in order to obtain the necessary sensitivity for many real-world analyses. Schemes such as solvent-solvent extraction and solid-phase extraction are often very tedious and time-consuming.
An alternative to these schemes is on-line preconcentration. In gas chromatography, this goal is met by passing a gas stream through a cold column that is subsequently heated. In high-performance liquid chromatography (HPLC), this process is usually done by gradient HPLC in which the analytes are retained on the column much more strongly for the first solvent than for succeeding ones. On-line preconcentration has also enjoyed some success in electrophoretic separations. For example, in capillary electrophoresis (CE), these include isotachophoresis, sample stacking, sweeping, and the use of a dynamic pH junction. In CZE, F. E. P. Mikkers, F. M. Everaerts, P. E. M. Verheggen, J. Chromatogr. 169 (1979), pp. 1-10 and R. L. Chien, D. S. Burgi, Anal. Chem. 64 (1992) pp. 489A-496A, demonstrated that changes in electric field strength between sample and background solution zones can focus (i.e., stack) charged species. In electrokinetic chromatography, J. P. Quirino, S. Terabe, Science, 282 (1998) pp. 465-68 and J. P. Quirino, S. Terabe, Anal. Chem. 71(8) (1999) pp. 1638-44, have shown that micelles can act to concentrate (i.e., sweep) neutral and charged species.
In CEC using particle (e.g., octadecyl silica) packed columns, focusing effects similar to that in gradient high performance liquid chromatography have been reported. These focusing effects were achieved using (1) step-gradient elution, (2) preparation of the sample in a noneluting solvent, or (3) injection of a water plug after sample injection. In M. R. Taylor, P. Teale, D. Westwood, D. Perrett, Anal. Chem. 69 (1997) pp. 2554-58, the authors were the first to report the use of a step-gradient for the preconcentration of steroidal samples in 1997. In D. A. Stead, R. G. Reid, R. B. Taylor, J. Chromatogr. A 798 (1998) pp. 259-67, the authors achieved a 17-fold increase in the detection sensitivity of a mixture of steroids by preconcentration using a noneluting sample matrix. In Y. Zhang, J. Zhu, L. Zhang, W. Zhang, Anal. Chem. 72 (2000) pp. 5744-47, the authors also used a noneluting solvent for the preconcentration of benzoin and mephenytoin by a factor of 134 and 219, respectively. In C. M. Yang, Z. El Rassi, Electrophoresis 20 (1999) pp. 2337-42, the authors reported on the preconcentration of a dilute sample of pesticides using a short plug of water injected after a long plug of sample. In M. J. Hilhorst, G. W. Somsen, G. J. de Jong, Chromatographia 53 (2001) pp. 190-96, the authors demonstrated preconcentration of structurally related steroids using a noneluting matrix and step-gradient elution. A gain in sensitivity of 7 to 9 times was reported. Similarly, in T. Tegeler, Z. El Rassi, Anal. Chem. 73 (14) (2001) pp. 3365-72, the authors just recently reported preconcentration of analytes in a mixture of carbamate insecticides using a combination of a noneluting matrix and step-gradient elution. The maximum allowable sample plug length was ˜20 cm and a 500-fold sensitivity increase is achieved for carbofuran. Zhang and co-workers achieved a further increase in detection sensitivity by combining field-enhanced sample injection with solvent gradient elution. They demonstrated a 17,000-fold increase in peak height for a positively charged analyte, propatenene.
It is desirable to provide a separation column with improved characteristics and that is easy to make.
A separation column includes a separation channel and a separation medium in the channel. The medium includes a porous matrix, and the porous matrix includes a support and a stationary phase. The support includes a metal organic polymer, and the stationary phase includes a bonded phase. The polymer may be a photopolymer. In the preferred embodiment, the porous matrix contains no chromatographic particles. The porous matrix may be used to both preconcentrate and separate analytes without chromatographic particles. The separation column allows for the concentration and separation of larger volumes of analytes than a separation column with chromatographic particles.
A method of preparing a monolith is provided. A separation column is provided. A mixture is introduced into the separation column. The mixture includes a metal organic compound. The mixture is irradiated to form a solid, porous matrix via photoinitiated polymerization. A coupling reagent is introduced into the column. Thus, a bonded phase porous matrix is formed in the column. In the preferred embodiment, the porous matrix is fritless and contains no chromatographic particles. Preparation of a fritless separation medium without chromatographic particles is simpler than preparing a separation medium with a frit or chromatographic particles. The porous matrix may be used both to preconcentrate and separate analytes.
The photochemical route to the preparation of the porous matrix has many advantages: (1) short preparation time, (2) control of the pore size, (3) control over the placement and length of the porous matrix, (4) high mechanical strength, and (5) avoidance of high temperatures that lead to cracking. An advantage of a bonded phase is the ability to change a number of conditions in chromatographic separations.
A method of separating a sample of analytes is provided. A separation column including a separation channel and a separation medium in the channel is provided. The medium includes a porous matrix, and the porous matrix includes a support and a stationary phase. The support includes a metal organic polymer, such as a photopolymer, and the stationary phase includes a bonded phase. In the preferred embodiment, the porous matrix contains no chromatographic particles. A sample of analytes carried in a solution is introduced through the column. The medium concentrates the analytes on the column. A solution is caused to flow through the column, thereby separating and eluting the analytes. The medium both preconcentrates and separates the analytes. In addition to the effect exerted by the medium, preconcentration can be further enhanced by a solvent gradient or sample stacking.
The above and other features and aspects of the present invention will become more apparent upon reading the following detailed description in conjunction with the accompanying drawings, in which:
For simplicity of description, like reference symbols are used for like or similar parts in this application.
Tube 12 can have many different cross-sections, such as a circular cross-section. Alternatively, tube 12 can have an elongated cross-section. These and other cross-sections are possible for tube 12 and are within the scope of the invention. Tube 12 may be a round capillary typically made of fused silica. The inside diameter (i.d.) of the capillary may be from about 10 μm to about 1000 μm, preferably from about 75 μm to about 500 μm. Tube 12 may be a planar chip or a confined space, such as a column confined by two sheets.
The medium 15 includes porous matrix, and the porous matrix includes a support and a stationery phase. The support includes a metal organic, or metalorganic, polymer, such as a photopolymer, wherein the term metal organic, or metalorganic, refers to a material that comprises an organic ligand attached to a metal atom or a metalloid atom, and the stationary phase includes a bonded phase. (See Brinker, C. Jeffrey, et al., Sol-Gel Science, The Physics and Chemistry of Sol-Gel Processing, p. 2 (1990).) A precursor of the polymer may be a metal alkoxide, wherein the term metal alkoxide refers to a metal organic, or metalorpanic, material, that has a metal-oxygen-carbon linkage or metalloid-oxygen-carbon linkage. (Id.) Herein, where "metal" is used in connection with a metal organic, or metalorganic, material, or in connection with a metal alkoxide, it encompasses metals and metalloids. The metal or metalloid may be aluminum, barium, antimony, calcium, chromium, copper, erbium, germanium, iron, lead, lithium, phosphorus, potassium, silicon, tantalum, tin, titanium, vanadium, zinc, or zirconium. The precursor may include a photoactive group, such as methacrylate. In one embodiment, the precursor may be trimethoxysilypropyl methacrylate, also known as methacryloxypropyltrimethoxy silane.
Different functionalized or derivatized monomers can be used to prepare a porous matrix with different physical properties, such as pore size, polymer charge density, and hydrophobicity. Control of the pore shapes and sizes through the use of different porogens can result in a porous matrix with a wide distribution of pore sizes (i.e., pore-size gradient). A porous matrix with a pore-size gradient can function as "molecule sorter" in capillary electrophoresis and capillary electrochromatography. The porous matrix can separate a mixture of large molecules whose size structures or chemistries (e.g., DNA fragments) may differ. In addition, separation columns 11 can be designed for reversed-phase, size-exclusion, affinity, ion-exchange chromatographies, etc.
A porous matrix may be a mixed phase porous matrix prepared from a mixture of monomers. For example, the monomers may include methacryloxypropyltrimethoxy silane, bis(triethoxysilyl)ethane, and bis(triethoxysilyl)octane. The mixed phase porous matrix may have different properties, such as hydrophobicity.
The properties of the porous matrix can also be changed by the use of a bonded phase. A bonded phase is a stationary phase which is covalently bonded to the support particles or to the inside wall of the column tubing. For example, the bonded phase may include a functional group bonded to a surface of the metal organic polymer. The functional group may be any functional group, such as a hydrophobic or hydrophilic group, a charged or uncharged group, or an organic or inorganic group. The functional group can alter properties of the porous matrix, such as the hydrophobicity, the charge, the retention factor, the type of interaction, or the chirality of the porous matrix. The bonded phase porous matrix can be used in normal phase, reversed phase, ion exchange, affinity, hydrophobic interaction, size-exclusion, or chiral chromatography.
A bonded phase can enhance the resolution of a separation of analytes and can deactivate to some extent the surface of the polymer. The deactivation decreases the surface activity caused by the existing negative charge from the ionization of the free hydroxyl groups. The bonded phase porous matrix can then support electroosmotic flow (EOF) in the column.
The bonded phase porous matrix has an affinity for analytes and can be used to both preconcentrate and separate a sample of analytes. The affinity for an analyte may be described by the retention factor, k, of the analyte. The retention factor, k, is equal to
The separation column 11 may be used for analytic or semipreparative work. Separation of analytes in the submilligram to milligram quantities may become possible with preconcentration on separation columns 11. For example, more than about 100-nL of sample solution at analyte concentrations in the mM levels can be injected into the column without significant evidence of overloading.
A method of preparing a monolith in the separation column 11 is provided. A separation column is provided. The separation column may be a round capillary typically made of fused-silica. The inside diameter (i.d.) of the capillary may be from about 10 μm to about 1000 μm, preferably from about 75 μm to about 500 μm.
A mixture that includes a metal organic compound is introduced into the separation column. The mixture forms a solid, porous matrix via photoinitiated polymerization. The mixture may comprise a metal organic monomer, a porogen, and a photoinitiator. The metal organic monomer may be a metal alkoxide, such as a silane, or a mixture of metal alkoxides. The metal may be aluminum, barium, antimony, calcium, chromium, copper, erbium, germanium, iron, lead, lithium, phosphorus, potassium, silicon, tantalum, tin, titanium, vanadium, zinc, or zirconium. The metal alkoxide may include a photoactive group, such as methacrylate. In one embodiment, the precursor may be trimethoxysilypropyl methacrylate, also known as methacryloxypropyltrimethoxy silane. In another embodiment, the precursor may be a mixture of methacryloxypropyltrimethoxy silane and another precursor, such as bis(triethoxysilyl)ethane or bis(triethoxysilyl)octane.
The metal organic monomer may be added to an acid or base catalyst for the hydrolysis of the precursor. The catalyst converts the alkoxy groups to hydroxyl groups. For example, a silane may undergo the following hydrolysis reaction to form a fully hydrolyzed silane:
A porogen or a mixture of porogens may be mixed with the metal organic monomer and the catalyst, and the mixture may be stirred for about zero minutes to about twenty-four hours. During this time, the metal organic monomer may undergo a condensation reaction to form dimers, trimers, and other oligomers. For example, a partially hydrolyzed silane may undergo the following condensation reaction:
The porogen provides a molecular template to form pores within the matrix. For example, the porogen may be a solvent, such as toluene or a 1:1 mixture of hexane and toluene, a polymer, or an inorganic salt, such as sodium chloride powder or sodium sulfate. The polymeric porogen may be poly(methyl methacrylate) or polystyrene, as reported by D. Horak, J. Labsky, J. Pilar, M. Bleha, Z. Pelzbauer, F. Svec, Polymer 34(16) (1993) pp. 3481-89. The porogen may be selected controllably to form pores in the matrix 15. The porosity of the matrix may be controlled by the type of chemical (i.e., porogen) used and its volume or concentration in the reaction solution. For example, a molar or volume ratio of monomer to porogen may be selected to form pores in the mixture. By adjusting the molar ratio of the monomer and porogen, the physical properties (e.g., pore sizes) of the matrix may be controlled.
The photoinitator or a photoactive group, such as methacrylate, on the monomer absorbs light to catalyze the polymerization of the metal organic compound. In one embodiment, the photoinitiator may be Irgacure 1800, available from Ciba Geigy, Tarrytown, N.Y.
If the separation column has an outer coating that is not transparent to the light source, the coating is first removed to make an irradiation window. The length of the coating will determine the length of the porous matrix formed within the separation column.
The mixture is introduced into the separation column. For example, the mixture may be flowed through the separation column using a syringe. The ends of the separation column may be sealed.
The mixture is irradiated, thereby forming a homogeneous separation medium in the separation column. For example, the separation column may be exposed to radiation for a short period of time, such as about five minutes. The wavelength of the radiation is dependent on the type of photoinitiator or photoactive group used in the reaction. The radiation may include visible or ultraviolet light For example, radiation of 365 nm may be used for the photoinitiator Irgacure 1800. The photoactive group methacrylate may be photopolymenzed at a wavelength of 300 nm or 365 nm, as reported in C. Yu, F. Svec, J. M. J. Frechet, Electrophoresis 21(1) (2000) pp. 120-27 and H.-G. Woo, L.-Y. Hong, S.-Y. Kim, S.-H. Park, S.-J. Song, H.-S. Ham, Bull. Korean Chem. Soc. 16(1995) pp. 1056-59, respectively.
A photochemical reaction occurs when the mixture is exposed to radiation. The photoinitator or photoactive group on the monomer absorbs the light to catalyze the polymerization of the metal organic compound.
The photochemical route to the preparation of a porous matrix has many advantages: (1) short preparation time, (2) control of the pore size, (3) control over the placement and length of the porous matrix, (4) high mechanical strength, and (5) avoidance of high temperatures that lead to cracking.
The separation medium preferably does not have frits or chromatographic particles, so the preparation of the separation medium is easier than preparation of a conventional separation medium with frits or chromatographic particles is.
Optionally, an organic solvent may be introduced into the separation column. For example, the organic solvent may be ethanol. The porous matrix may be washed clean of any unreacted material, the porogen, and the photoinitiator using an organic solvent. The solvent may be flowed through the separation column using a syringe or other means.
A coupling reagent is introduced into the column to form a bonded phase porous matrix. The coupling reagent may have monofunctionality, difunctionality, or trifunctionality. The coupling reagent may be an organic coupling reagent, such as a halogenated aromatic or an acyl halide. For example, sodium cyanide and ammonium chloride can react with a ketone on the metal organic polymer, or a halogenated aromatic or an acyl halide can undergo a condensation reaction. The coupling reagent may include a functional group and a metal. The metal may be aluminum, barium, antimony, calcium, chromium, copper, erbium, germanium, iron, lead, lithium, phosphorus, potassium, silicon, tantalum, tin, titanium, vanadium, zinc, and zirconium. For example, a silane coupling reagent may be an organochlorosilane, an organoalkoxysilane, an organoaminosilane, or other reactive silanes. The silane coupling reagent can be used to derivatize a surface hydroxyl group of the metal organic polymer with a functional group. For example, the metal organic polymer and a silane coupling reagent, such as an organochlorosilane, including a functional group, R′, may undergo the following reaction to form a bonded phase porous matrix:
Optionally, an organic solvent may be introduced into the column including the bonded phase porous matrix. For example, the organic solvent may be toluene, pyridinium, or triethylamine. Any unreacted silane coupling reagent and byproducts of the reaction, such as hydrochloric acid or an alcohol, may be removed from the bonded phase porous matrix by introducing the organic solvent into the column. The solvent may be flowed through the separation column using a syringe or other means.
The separation column may be conditioned with a separation solution before using the column for separation. A separation solution may comprise a buffer, such as aqueous ammonium acetate, and an eluting solvent, such as acetonitrile.
A method to separate a sample of analytes is provided. A sample of analytes is introduced in a sample solution through a separation column 11. Analytes includes neutral species, such as polycyclic aromatic hydrocarbons, alkyl benzenes, alkyl phenyl ketones, and steroids, and charged species, such as peptides. The sample solution may comprise a buffer, such as aqueous ammonium acetate, and an eluting solvent, such as acetonitrile.
The separation column 11 includes a separation channel 13 and a homogeneous separation medium 15 in the channel 13. The separation column 11 may be a capillary or planar structure. The separation medium 15 preferably does not have frits or chromatographic particles. The use of a homogeneous separation medium is advantageous because, in some applications, the use of chromatographic particles (i.e., inhomogeneous separation phase) introduces unwanted broadening (i.e., lack of resolution). The separation medium 15 may possibly include two sections containing substantially the same porous homogeneous stationary phase, and the two sections may be separated by another section, such as a monolith with a different pore size or surface charge. Preferably, the separation medium 15 is continuous.
The separation medium 15 includes a porous matrix, and the porous matrix includes a support and a stationary phase. The support includes a metal organic photopolymer, and the stationary phase includes a bonded phase. The separation medium 15 concentrates the analytes on the column. The sample may be introduced by applying a pressure or a voltage to the column 11. For example, the pressure may be 0.5 psi or 20 psi for a period of time, such as two to 1920 seconds, or a field strength of about 40 V/cm may be applied for a period of time. For example, an injection plug length of greater than about two centimeters may be injected into the column 11.
The analyte is concentrated by the column 11. The extent of preconcentration is purely dependent on the retention factor, the k value. The retention factor is affected by the nature of the solvent, the nature of the analyte, and the detailed morphology of the separation medium. Moreover, the flow rate hardly influenced the extent of preconcentration.
The highly porous nature of the porous matrix results in a high mass transfer rate for the analyte, which facilitates the preconcentration effect. The high mass transfer rates arise from the enhanced accessibility of the analytes to the binding sites of the porous structure. Because of the high mass transfer rates, the kinetics of analyte-porous matrix interaction (i.e., the partitioning of the analyte between the mobile and stationary phases) is not the rate-limiting step in the separation. The high mass transfer rate distinguishes this separation method from previous forms of chromatographic separations. With this separation method, because of the high mass transfer rate, it is possible to inject and concentrate larger volumes of sample solution than in columns containing normal chromatographic materials.
The total preconcentration effect is directly proportional to the retention factor, with longer injection plug lengths (e.g., greater than about 25 mm) leading to severe peak broadening of analytes having low-k values. This behavior implies a maximum length of sample plug for each analyte before peak shape becomes compromised. The retention factor can be altered by changing the bonded phase.
A major advantage of on-line preconcentration is that it lowers the detection limit for a given analyte. Another advantage is that preconcentration may be used to clean up the analytes from possible interfering species found in the sample matrix.
A separation solution is caused to flow through the column 11, thereby separating and eluting the analytes. The separation solution may be caused to flow by applying a voltage or a pressure to the column 11. For example, the pressure may be 0.5 psi or 20 psi for a period of time, such as two to 1920 seconds, or a field strength of 300 V/cm may be applied for a period of time. The separation solution may comprise a buffer, such as aqueous ammonium acetate, and an eluting solvent, such as acetonitrile. In one embodiment, the separation solution is the same as the sample solution.
The bonded phase porous matrix acts to extract the analytes from solution as well as provides the stationary phase for chromatographic separation of the analytes. It is this extractor-separator combination that gives this method such power. For example, more than about two-centimeter plugs of sample solution can be loaded into the capillary and concentrated using a separation solution that is the same as the sample solution.
In one embodiment, the analytes may be separated by performing normal phase, reversed phase, ion exchange, affinity, hydrophobic interaction, size-exclusion, or chiral chromatography.
In another embodiment, in addition to the effect exerted by the porous matrix, a solvent gradient may be used to further enhance preconcentration of the analytes. In this embodiment, the sample may be dissolved in a solution with a higher concentration of a buffer (e.g., water) than in the separation solution. The higher concentration of the buffer in the sample solution increases the affinity of the sample to the stationary phase. When a solvent gradient is used, the plug length can be longer than the length of the column. For example, injection of a 91.2-cm plug, which was more than three times the total length of the capillary, was possible with only a minor loss in resolution. Improvements in peak heights obtained under gradient conditions can be more than a thousand-fold.
For neutral analytes, two approaches exist for using gradients on the porous matrix. The first approach is to increase the organic solvent ratio between the separation solution and the sample solution. The second approach is to increase the retention factor k in the separation by increasing the percentage of water in the separation solution while maintaining a reasonable percentage of organic solvent between the separation solution and the sample solution. Analysis is faster with the first approach, whereas the resolution is better with the second one.
In another embodiment of the present invention, in addition to the effect exerted by the porous matrix, sample stacking may be used to further enhance the preconcentration of analytes. Sample stacking is the focusing of charged analytes when analytes pass the concentration boundary that separates regions of high and low electric field strengths. The high electric field zone is a lower conductivity sample solution containing more of the eluting solvent, whereas the low electric field region is a higher conductivity separation solution. The eluting solvent, such as acetonitrile, has a lower conductivity than the buffer, such as aqueous ammonium acetate. Thus, a higher concentration of the eluting solvent results in lowering the sample matrix conductivity.
The separation column is prepared with the separation solution. When analytes are introduced into the separation column and a voltage is applied, the analytes in the sample solution at the inlet of the column rapidly accelerate toward the separation solution (lower electric field strength) already in the column, where on crossing the boundary between the sample solution and the separation solution, they slow down and stack into narrow zones at the interface.
Sample stacking is basically caused by the change in electrophoretic velocity at the concentration boundary. Electrophoretic velocity is the product of electrophoretic mobility and electric field strength. Focusing occurs (sample stacking) when the electrophoretic velocity decreases at the concentration boundary. Sample stacking is also explained using the fundamentals of isotachophoresis and Kohlrausch rules.
There are two approaches to perform sample stacking on a porous matrix. The first approach is to increase the percentage of organic solvent, such as acetonitrile. The second is to decrease the concentration of the buffer component in the sample solution. Increasing the percentage of acetonitrile or other suitable organic solvent is especially useful for real samples containing high concentration of salts. Desalting, for example by dialysis, is therefore not necessary to make a lower conductivity solution for injection. Use of organic solvents is also useful for biological samples when deproteination is part of the sample preparation.
The invention is described in more detail by the way of the following examples. The following examples are presented solely for the purpose of further illustrating and disclosing the present invention, and are not to be construed as limiting the invention.
Materials and Chemicals. Fused-silica capillaries (75-μm i.d.�365-μm o.d.) were purchased from Polymicro Technologies, Phoenix, Ariz. Methacryloxypropyltrimetoxysilane (MPTMS) was purchased from Gelest, Tullytown, Pa. and Sigma-Aldrich, Milwaukee, Wis. and was used without purification. HPLC-grade toluene, phenanthrene, pyrene, alkyl benzenes, alkyl phenyl ketones, and steroids were purchased from Signa-Aldrich, Milwaukee, Wis. Irgacure 1800 was received from Ciba, Tarrytown, N.Y.
Instrumentation. A Beckman P/ACE 2000 capillary electrophoresis instrument with a UV-absorbance detector was used to carry out all CEC experiments. An XL-1500 UV cross-linker, available from Spectronics Corp., Westbury, N.Y., equipped with six 15 W blacklight tubes of predominantly 365-nm wavelength was used to irradiate the reaction solutions. Scanning electron microscopy (SEM) analyses were performed on a Philips SEM 505 scanning electron microscope, available from Eindhoven, Netherlands.
Polymerization Procedure. The monomer stock solution was prepared just prior to use by adding 375 μL of MPTMS to 100 μL of 0.12 N HCl. This solution was stirred at room temperature for approximately thirty minutes to afford a clear, monophasic solution. An appropriate amount of toluene porogen) was added to the monomer stock solution, as shown below in Table 1.
A 15-cm stripe of the polyimide coating on a 30-cm long capillary was removed using a razor blade positioned at 45� to the capillary surface. The mechanical stability of the capillary was remarkably good despite the removal of a stripe of polyimide coating. The irradiation light entered the capillary only through this 15-cm stripe. No monolith was formed in the capillary where the polyimide coating ("mask") remained intact.
Using a 0.5-mL disposable syringe, approximately 0.2 mL of the reaction solution was flushed through the capillary to wet thoroughly the wall surface before filling the capillary with the solution. This resulted in bonding of the monolith to the capillary wall. No special pretreatment of the capillary wall was necessary to bond the monolith to the wall. The filled capillaries were irradiated (900 mJ/cm2) in a UV cross-linker using 365-nm light for five minutes to form the porous matrix.
After irradiation, the capillaries were washed with ethanol using a hand-held syringe to removed unreacted reagents or porogens. Because the monoliths were highly permeable, high pressures were not required to drive liquid through the capillaries. Once the unreacted reagents were removed, the monolith became opaque and could be viewed clearly through the capillary without the aid of a microscope. The homogenity of the porous matrix was confirmed at 100� magnification. Burning off the polyimide coating immediately after the monolith section with fuming sulfuric acid made a detection window.
Once fabricated, the capillary was successfully installed in the cartridge without any damage. The capillary was conditioned with the separation buffer for approximately five minutes using a syringe and a hand-held vise. Once in the instrument, the capillary was further conditioned by pressure rinsing (20 psi) with the separation buffer or by electrokinetically conditioning at 5 kV or 10 kV for thirty minutes.
Characterization. SEM was used to study the morphology of the separation column. A capillary was sectioned carefully to expose the monolith. The sectioned pieces of capillary were sputtered with gold prior to SEM analyses.
Analyte Separation. The analytes were prepared in the mobile phase to prevent gradient effects during the CEC experiments. The mobile phase was made up of various ratios (v/v) of 50 mM ammonium acetate, water, and acetonitrile. A new sample solution was used for every injection to maintain the same concentration of acetonitrile in the sample solution and the mobile phase.
The elution order of the column was similar to that of reversed-phase chromatography with the larger molecular weight or more hydrophobic analytes eluting later than the smaller molecular weight or more hydrophilic analytes. Elution of the analytes in both figures occurred in less than seven minutes. Bubble formation was not a problem during the CEC experiments, for which the typical operating currents were between 3 and 10 μA.
For a typical capillary column D, efficiencies of up to 100,000 plates/m are achieved for thiourea, a less-retained compound. Small variations in the elution times were observed for thiourea (0.65% RSD), naphthalene (1.10% RSD), phenanthrene (1.14% RSD), and pyrene (1.14% RSD) over a period of three days (n=33).
The permeability of a porous matrix was determined by the linear velocity of the porous matrix, which is proportional to permeability as described in Darcy's law. The permeability of a porous matrix as a function of the macropore size was highly dependent on the volume and type of porogen used to prepare the photopolymer. For a column made with 90% (v/v) toluene (column A), the linear velocity is 12.3 cm/min, and an 80% (v/v) column (column B) had a linear velocity of 3.3 cm/min, whereas a column made with 73% (v/v) toluene (column D) had a linear velocity of 0.6 cm/min. These linear velocity data suggested that the macropores decrease with decreasing porogen concentration. This behavior was consistent with what has reported in the literature.
The separation column was prepared as described in Example 1. A mixture of 1:1 hexane/toluene was used for the porogen. The separation column had a separation performance similar to that of a separation column made with 80/20 toluene/reaction solution. A column efficiency of 68,000 plates/m (RSD 7.0%, n=5) for thiourea and an electroosmotic flow (EOF) velocity of 3.7 cm/min was obtained.
A mixture of 375 μL of MPTMS and 100 μL of 0.12 M hydrochloric acid was stirred for thirty minutes at room temperature. 27 parts of this mixture were combined with 73 parts of toluene to give 200 μL of the final solution. 5% by weight of the final solution of the photoinitator Irgacure 1800 was added, and the resulting sol-gel solution was stirred for five minutes before use. A 75-μm i.d.�365-μm o.d. fused silica capillary was filled with the sol-gel solution, and the separation column was exposed to UV light in a Spectrolinker X1-1500 at 365 nm to affect photopolymerization. The polymerization length of the porous matrix was controlled by removing a 15-cm strip of the polyimide coating of the capillary prior to irradiation for five minutes. Unreacted reagents were flushed with ethanol. The total length of the capillary was 25.6 cm (18.8 cm from inlet to the detector window). The resulting column was conditioned with the separation solution prior to use.
All electrophoresis experiments were performed with a Beckman P/ACE 2000. The capillaries were thermostated at 20� C. Injections were done using pressure (i.e., 0.5 psi and 20 psi) or voltage (1 kV to 10 kV) and varied in duration from two seconds to 1920 seconds. Detection was done at 214 or 254 nm. Data analysis was performed with GRAMS/32 version 4.02, available from Galactic Industries Corporation, Salem, N.H.
For the PAH mixture illustrated in
The separation column was prepared as described in Example 3. The sample and separation solution was 50 mM ammonium acetate/water/acetonitrile (1/3/6). The samples were injected at 1 kV. The applied voltage was 15 kV, and the absorbance was detected at 214 nm.
Preconcentration was evidenced by the slightly higher peak height for the longer injection of diluted sample and almost the same corrected peak widths (peak width/migration time) for both experiments, despite the different sample concentration. The peak heights of the electrochromatograms in signals a and b were 0.0869 (% RSD=0.36%, n=3) and 0.0937 (% RSD=0.06%, n=3) arbitrary units, respectively. The peak widths of the electrochromatograms in signals a and b were 0.0253 (% RSD=0.07%, n=3) and 0.0249 (% RSD=0.01%, n=3) arbitrary units/min, respectively. The shift in migration time on line b was caused by the longer injection time, which made the center of the sample plug closer to the detector window.
A mixture of 575 μL of MPTMS and 100 μL of 0.12 M hydrochloric acid was stirred for thirty minutes at room temperature. 20 parts of this mixture were combined with 80 parts of toluene to give 200 μL of the final solution. The photoinitiator was added as 10% of the total volume of the final solution, and the resulting sol-gel solution was stirred for five minutes before use. The separation column was prepared as described above in Example 3. Unreacted reagents were flushed with toluene. The surface of the porous matrix was modified by continuous flow of pentafluorophenyltrichlorosilane through the capillary for forty-five minutes at room temperature and followed by rinsing with toluene.
The separation column was prepared as in Example 3.
The separation column was prepared as described in Example 3.
The retention factors, k, obtained for acetophenone (peak 2), propiophenone (peak 3), butyrophenone (peak 4), valerophenone (peak 5), hexanophenone (peak 6), heptanophenone (peak 7), octanophenone (peak 8), and decanophenone (peak 9) were 0.18, 0.25, 0.32, 0.41, 0.53, 0.67, 0.85, and 1.33, respectively. In this study, thiourea (peak 1) was used as the essentially unretained neutral solute for the determination of k. The value of k and migration time follows the increase in alkyl chain length. In general, the peak shapes and resolution improved when the water concentration was increased from 40% to 50%, 60%, 70%, and 80%, as evidenced by panels a, b, c, d, and e, respectively.
The experimentation conditions were the same as in Example 7.
The separation column was prepared as described above in Example 3.
The gradient condition, as shown in
The k values were higher in
A solvent gradient improved detection of four PAHs, as shown in
Improvements in peak heights for naphthalene (peak 2), phenanthrene (peak 3), pyrene (peak 4), and benz(e)acephenanthylene (peak 5) are 346, 437, 409, and 315 times, respectively. The sample concentrations in
The peak heights of decanophenone (peak 2) and pyrene (peak 3) increased with increasing plug lengths. The injection was increased from 0.23 mm (panel a) to 7.6 cm (panel b), 0.22.8 cm (panel c), 45.6 cm (panel d), and 91.2 cm (panel e), which corresponds to 0.1%, 30%, 89%, 178%, and 356% of the total capillary length. The high porosity or the low resistance to flow of the porous matrix made it possible to introduce increasing lengths of the sample solution in a rather effortless manner. Longer than 91.2 cm injection is still possible. It is not performed, however, owing to loss of resolution as observed in
Successful separations have been done with PSG in 250-μm i.d. capillaries (data not shown). This work opens the possibility of performing semi-preparative separations involving long plug injections. Injection volumes in the μl range could easily be made.
Although it was expected that the peak shapes would be better under a gradient condition (
Undesirable peak shapes are observed in
The analytes in
Field enhanced sample injection was performed by dissolving the sample in a low conductivity matrix (0.5 μM phosphoric acid in 40% acetonitrile), followed by injection using voltage with the negative electrode at the detector end. As the voltage was applied, the low conductivity sample matrix entered the capillary by virtue of electroosmotic flow (EOF) while the cationic peptides entered the column by virtue of both EOF and electrophoretic flow. Only a very small plug of sample matrix was introduced because the low pH of the separation solution markedly decreases the EOF, which prevents the dissociation of silanol groups at the capillary walls. An unretained neutral solute (thiourea) was actually detected after 30 minutes.
The electric field in the sample matrix zone introduced into the column was much higher than the separation zone. This effect caused the high electrophoretic velocity of the cationic peptides entering the capillary. The high analyte electrophoretic velocity caused a large amount of peptides to be introduced, unlike in hydrodynamic injection, the volume of sample loaded limited the amount of sample introduced The high analyte electrophoretic velocity also caused focusing or preconcentration of peptides at the concentration boundary between the sample matrix and separation solution (sample stacking). Introduction of a water plug before electrokinetic injection, which is suggested to be useful in sample stacking with electrokinetic injection, did not improve the peak heights because of the similar direction of the EOF and analyte electrophoretic velocities. The low conductivity sample matrix that entered the capillary also maintained the enhancement of the electric field at the inlet end of the capillary during injection.
With the conditions in
Table 2 lists the types and volumes of reagents used to make different precursor stock solutions where the ratio of the acid catalyst to the precursor, methacryloxypropyltriethoxysilane, was varied or where the precursor was reacted with a co-precursor (to form a mixed phase PSG monolith).
Either changing the concentration of the precursor in the reaction solution or using a co-precursor for the formation of mixed phases modified the chemical nature of the parent PSG monolith. The PSG monoliths, PSG-A and PSG-J, prepared from solutions A and J, respectively, differ only in the volume of precursor used in the reaction with J containing a higher volume of the precursor than A. A higher volume of the precursor in the reaction should result in a denser monolith in the capillary column. The PSG monolith, PSG-K, was prepared with the precursor and bis(triethoxysilyl)ethane as a co-precursor. The PSG monoliths, PSG-M and PSG-P, were prepared with the precursor and different amounts of bis(triethoxysilyl)octane as the co-precursor. The co-precursors hydrolyze and condense with the precursor to form hybrid sols (mixed phases).
For solutions A and J, the appropriate volume of the precursor was added to 100 μL of the acid catalyst (0.12 M HCl), and the resulting solution was stirred for 15 minutes at room temperature (in the dark). For solutions K, M, and P, the appropriate volume of the precursor was added to 100 μL of 0.12 M HCl followed by the addition of the appropriate amount of bis(triethoxysilyl)ethane; the resulting solution was stirred for 15 minutes at room temperature (in the dark). All of these solutions were used within two hours of their preparation.
A similar procedure was followed in making the toluene/precursor stock solutions with the photoinitiator added. The amount of photoinitiator added to the toluenelprecursor stock solution was 10 mg photoinitiator for every 100 μL of the toluene/precursor stock solution.
The capillary was prepared and conditioned in the same manner as previously described. PSG capillary column conditioning was the same as before.
The separation factors of the monoliths for two test mixtures of alkyl phenyl ketones (APKs) and polycyclic aromatic hydrocarbons (PAHs) were determined. The separation factor is a measure of the analyte separation capability of a chromatographic system. The separation factor, a, is given by k2/k1, where k is the retention factor for a particular analyte, and k2 and k1 are the k values for adjacent analytes. The retention factor k=(tr−to)/to was determined in the usual way, where tR is the analyte retention time and to is the retention time of an unretained marker, for which we used thiourea. Table 3 lists the separation factor of each PSG monolith for naphthalene and pyrene.
The values for a varied from 2.43 for PSG-K to 2.76 for PSG-P with the separation factor for PSG-P being slightly higher than that of the other monoliths. Separation factors greater than 1 indicated successful separation of the analytes.
Resolution was determined from the expression
The use of the co-precursor, bis(triethoxysilyl)octane in PSG-M and bis(triethoxysilyl)ethane in PSG-K and PSG-P resulted in resolution for naphthalene and pyrene of 4.37, 4.09, and 9.03, respectively, which is an enhancement of up to 73% as compared to the resolution on the parent PSG-A (Rs=2.43). The retention factors for naphthalene (0.23) and pyrene (0.56) were both 60% higher for these three monoliths than for PSG-A.
The separation column was prepared as described above in Example 15 for monolith PSG-J. For a porous matrix having a length of 15 cm, the retention factors for napthalene and pyrene, kN and kpy, respectively, were 0.31 and 0.79, respectively, for a porous matrix made with 80% toluene. For a similar porous matrix having a length of 10 cm, kN and kpy were 0.10 and 0.24, respectively. There was a linear relationship between length and kN (r=0.991) and kpy (r=0.991). The separation factors for 15-cm, 10-cm, and 5-cm porous matrices were 2.55, 2.52, and 2.40, respectively. Thus, for the shortest monolith length, a high separation factor was maintained, while the elution times for the analytes were significantly reduced. Decreasing the length of the porous matrix in a capillary column led to a decrease in the elution times of the test analytes. Decreasing this length had an effect of decreasing the retention factors of naphthalene and pyrene.
The separation column was prepared as described above in Example 15. For PSG-A made with 80% toluene, kN was 0.14 and kpy is 0.36, whereas kN was 0.30 and kpy was 0.74 for PSG-A made with 73% toluene. The separation factors for PSG-A made with 80% toluene and 73% toluene were 2.57 (0.1% RSD) and 2.47 (0.1% RSD), respectively. The value of k increased by 53% and 51% for naphthalene and pyrene when 73% toluene was used in the preparation of the monoliths.
A similar trend was observed for PSG-J where kN was 0.31 and kpy was 0.79 for a monolith made with 80% toluene. The kN (0.49) and kpy (1.23) values increased by 37% and 36% when the concentration of toluene was decreased from 80% to 73%. The separation factors of PSG-J made with 80% toluene and 73% toluene were 2.55 and 2.51, respectively.
The resolution of naphthalene and pyrene differed significantly when comparing PSG monoliths made with 80% and 73% toluene. When the pore size decreased, which was brought about by using lower volumes of toluene, the PSG surface increased with a resulting increase in the retention and resolution under the same separation solution conditions. Thus, the permeability of the porous matrix affects the retention of the analytes.
The monomer, methacryloxypropyltrimethoxysilane (MPTMOS) and the silane coupling reagents, pentafluorophenyldimethylchlorosilane (PFPDM), pentafluorophenyltriethoxysilane (PFP), 3,3,3-trifluoropropyltrichlorosilane (C3F3), n-octadimethylchlorosilane (C8), (tridecafluoro-1,1,2,2-tetrahydrooctyl) dimethylchloro-silane (CF13), and n-propylaminotriethoxysilane (NH2), were purchased from Sigma-Aldrich (Milwaukee, Wis.) or Gelest (Tullytown, Pa.) and used as received. The photoinitiator, Irgacure 1800, was donated by Ciba (Tarrytown, N.Y.). All solvents were of spectroscopic grade from Sigma-Aldrich. Fused-silica capillaries of 75-μm inside diameter �365-μm outside diameter were purchased from Polyrnicro Technologies Phoenix, Ariz.). Thiourea, alkyl phenyl ketones, naphthalene, phenanthrene, pyrene, nucleosides, taxol, taxol derivatives, and the peptides were purchased from Sigma-Aldrich and used as received.
All of the electrochromatograms were obtained using a Beckman P/ACE 2000 capillary electrophoresis (CE) instrument equipped with a UV absorbance detector (Beckman Coutler, Inc. Fullerton, Calif.). A Spectrolinker XL-1500 equipped with six 365 nm low-pressure mercury bulbs (Spectronics Corp., Westbury, N.Y.) was used in the photopolymerization reactions. A Philips Model 505 scanning electron microscope (SEM) was used to analyze the physical suture of the porous matrices in the capillary columns. For that purpose each column was coated with a 10-nm thickness of gold/palladium using a Denton Vacuum, LLC Desk II Cold Sputter/Etch unit and carbon evaporation accessory (Moorestown, N.J.).
The metal organic polymer was prepared with a monomer stock solution of 575 μL MPTMOS and 100 μL of 0.12 M HCl that was stirred at room temperature for 15 minutes. An 80/20 toluene precursor solution (v/v) was prepared by mixing 60 μL of the monomer stock solution with 30 mg of the photoinitiator dissolved in 240 μL of toluene. The resulting solution was stirred at room temperature and in the dark for 5 minutes. A capillary column with a 5 or 10-cm stripe of polyimide removed from its exterior was prepared. The total length of the capillary was 25.6 cm (18.8 cm from inlet to the detector window). The detector window, which was positioned after the porous matrix, was created by burning off the polyimide with fuming sulfuric acid.
The bonded phase was prepared on an 80/20 metal organic polymer in a column that was rinsed with toluene prior to the silanization reaction. The rinsed column was then treated with the silane coupling reagent by continuous flow of a neat solution of the reagent with a syringe in a hand-held vise. The reagent was allowed to react with the surface of the polymer for 15, 30, 60, 70, and 90 minutes at room temperature. Any unreacted silane coupling reagent was removed from the bonded phase porous matrix by flushing with toluene using a syringe. The following bonded phase porous matrices were prepared by this procedure: PSG-PFP, PSG-PFPDM, PSG-C8, PSG-C3F3, PSG-CF13, and PSG-NH2.
A PSG capillary column was carefully installed into a P/ACE cartridge. Although the column is without a complete coating of polyimide, it still maintains good mechanical strength. Care is taken, however, in handling these capillaries to prevent breakage during installation into the capillary cartridge. Once in the cartridge, the capillary is first conditioned with the separation solution using a syringe. In the CE instrument the column is rinsed further for about 2 minutes at 20 psi followed by electrokinetic conditioning at 5 kV for 10 minutes. The columns were thermostated at 20� C.
The bonded phase porous matrix was prepared as described above in Example 18. The alkyl phenyl ketone sample solution was prepared with 1.42 μM acetophenone, 25 μM propiophenone, 15 μM butyrophenone, 20 μM valerophenone, 5.5 μM hexanophenone, 16.5 μM heptanophenone, 15.3 μM octanophenone, and 3.5 μM decanophenone in a solution of 50 mM ammonium acetate (pH 6.5)/water/acetonitrile (1/3/6). Using a lower volume of acetonitrile resulted in lowering the solubility of the sample. The separation solution was 1/5/4 50 mM ammonium acetate (pH 6.5)water/acetonitrile. Injection was at 1 kV for 5 seconds, and the separation voltage was 20 kV.
A comparison of the five bonded-phase PSG columns, namely PSG-C3F3 (panel b), PSGCF13 (panel c), PSG-PFP (panel d), PSG-C8 (panel c), and PSG-PFPDM (panel f), to the parent PSG column (panel a) revealed enhanced resolution of the test analytes, particularly for the more hydrophobic ones, with the highest enhancement occurring for PSG-PFPDM. Resolution was determined from the expression
It has been reported that fluorinated bonded phases interact with aromatic compounds based on electrostatic interactions between the π electrons of the aromatic rings and the lone pair electrons (in the 2p orbital) on the fluorine atom of the bonded phase. This behavior was not the case for the fluorinated bonded phases, PFP and PFPDM. Separation of the alkyl phenyl ketones was caused by the increasing hydrophobicity of the compounds as the number of carbons of the alkyl functional group increases. The elution order followed this increase in the hydrophobicity for these compounds.
PSG-C8 showed good resolution for the alkyl phenyl ketones and shorter analysis times for all eight alkyl phenyl ketones with decanophenone eluting within 20 minutes (panel e). PSG-C8 combined high resolution with fast elution times for the alkyl phenyl ketones and had an EOF of 1.76�10−4 cm2/Vs, which was 15% higher than the EOF values for the other bonded phase porous matrices, but the same as that of the underivatized porous matrix.
We observed no bubbles or drying out of the PSG columns, even at high field strengths employed in the separation of the alkyl phenyl ketones. The bonded phase porous matrices were more stable to highly acidic conditions (pH 24) than the underivatized porous matrix. A plot of the current versus field strength for each of the bonded phase porous matrices was linear (data not shown). This linearity suggests the absence of Joule heating, which can lead to peak broadening. In each of the separations, the efficiency of the alkyl phenyl ketones decreased with increasing k values. Diffusion effects that caused the analytes to slowly elute off the porous matrix may explain the low efficiencies. Higher resolution can be attained with a bonded phase porous matrix having a length of 15 cm, but at the expense of longer elution times. A linear relationship existed between the length of the porous matrix and k. The separation factors for bonded phase porous matrices of lengths 15, 10, and 5 cm were 2.55, 2.52, and 2.40, respectively.
Table 4 presents the effect of the silanization reaction time on the retention factor k, separation factor a, and resolution Rs of PSG-C8 for acetophenone (1) and hexanophenone (5) on a 5-cm PSG-C8 monolith.
Collapse of the bonded phase may explain the decrease in the resolution for reactions times of 70 and 90 minutes. As the silanization reaction time was increased, more of the n-octyldimethylsilane was allowed to bond to the hydroxyl groups of the metal organic polymer, thereby increasing the number of n-octyldimethyl groups on the polymer surface. As more of the bonded phase was bound on the polymer surface, the closer the ligands get to one another. The fact that the k1 and k5 values increased by 17% and 58% respectively, from 0 to 60 minute reaction time indicated that the polymer surface was becoming more hydrophobic, i.e., more retentive for the alkyl phenyl ketones.
The nucleoside sample mixture consisted of inosine, uridine, guanosine, and cytidine in concentrations of 1 mg/mL each in water. The separation solution was 50 mM phosphate (pH 8)/water/acetonitrile (0.5/3.5/6). The injection plug length was 0.1 mm, and the applied voltage was −15 kV.
A 10-cm PSG-NH2 monolith was prepared as described above in Example 18. A 10-cm long Porous matrix was derivatized with n-propylaminotriethoxysilane. The resulting monolith, PSG-NH2, had an amine-bonded phase (pH 9-10) that was positively charged under our experimental conditions. The positive charge allowed for reversal of the EOF. Although chromatographic materials with an amine group functionality had found some application in normal phase and ion-exchange chromatography, PSG-NH2 under our conditions behaved as a reversed phase material. The alkyl phenyl ketones eluted in the same order (data not shown) as compared to the other bonded phase porous matrices described above.
A 15-cm PSG-PFP monolith was prepared as described above in Example 18. Short chain peptides, angiotensin I, bradykinin, angiotensin II, gly-gly-gly, val-tyr-val, and methionine enkephaline, were used as cationic analytes. The concentration of each of the peptides was 16 μg/mL. The separation solution was 50 mM ammonium acetate (pH 4.3)/water/acetonitrile (1/4/5). Injection was for 30 s at 0.5 psi, and the applied voltage was 10 kV.
A 15-cm PSG-C3F3 monolith was prepared as described above in Example 18. Taxol, baccatin III, and acetylbaccatin were each prepared as 1 mg/mL solutions in acetonitrile. The separation solution was 50 mM ammonium acetate (pH 6.5)/water/acetonitrile (1/4/5). Injection was for 5 s at 0.5 psi, and the applied voltage was 10 kV.
While the invention has been described above by reference to various embodiments, it will be understood that changes and modifications may be made without departing from the scope of the invention. All references referred to above are incorporated herein by reference in their entireties.
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text: Publications Search Results Users may wish to consult What's Included in the Bibliography. Results 1 - 2 Results per page: 25 50 100 Sort by: Pub Date (newest) Title Recent Additions Author Clear All Pub. Year Citation 2003 Udry, J.R., Li, R.M., Hendrickson-Smith, J. Health and Behavior Risks of Adolescents with Mixed-Race Identity. American Journal of Public Health. 93, (11), 1865-1870. Full Text Options: DOI Worldcat Google Scholar Export Options: RIS EndNote XML 2002 Udry, J.R., Hendrickson-Smith, J., Li, R.M. . Health Behavior Risk for One-Race and Mixed Race Adolescents. Add Health Users Workshop. Bethesda, MD. Export Options: RIS EndNote XML [1]

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We can't guarantee that wearing the Mavic Sprint Shorts will take all the pain out of your quest for the finish line, but they'll certainly keep you sitting in comfort, and looking good doing it. After all, they've got a slim cut, a sensible chamois, and fabrics that work hard to keep you dry when you're laying it all on the line. And in case you're wondering, they're equally at home on your day-to-day rides as well.
To augment the pre-curved cut, Mavic employed its Wick Matrix ST fabric. Like the name implies, it's designed specifically to maximize breathability, pulling as much moisture as possible away your your skin to keep you dry. The flatlock seams were chosen to fight chafing, further adding to your comfort.
For the chamois, Mavic chose its Ergo 3D insert. This proven chamois uses three different densities of foam, with the firmest being underneath your sit bones. The result is a better dispersion of your weight as compared to a single density chamois, and that translates into fewer hot spots and better bloodflow.


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The Tampa Bay Buccaneers have traded wide receiver Arrelious Benn. Per the team's release, along with Benn is a seventh-round pick in the 2013 NFL Draft to the Philadelphia Eagles in exchange for a sixth-round pick in the 2013 NFL Draft and a conditional selection in the 2014 NFL Draft. The Buccaneers currently own eight draft choices in the upcoming draft, one each in rounds one, two, three and five and two in rounds four and six. Benn has been plagued with injuries since being drafted in the second round of the 2010 draft. He had 59 receptions for 862 yards and five touchdowns in three seasons with Tampa Bay. The post Buccaneers have traded Arrelious Benn to Philadelphia appeared first on The Bay Cave.
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i did a bunch of rough thumbs, which im posting, and i have one basic drawing down. I need to know how my perspective is?
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Gibson, AP; Tidswell, T; Bayford, RH; Holder, DS; (2000) Realistic and spherical head models in EIT. In:
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Terry Costa Hosts Bridal Event
Landmark Dallas boutique, Terry Costa, announced its 2013 Spring Bridal Fair will run today, January 5th-6th at the flagship location in Dallas. Cake samples will be available and Terry Costa's trusted local bridal vendors will be on-hand for demonstrations, presentations and giveaways.
"This is the perfect kickoff to bridal season -- so many women get engaged during the holidays and are instantly overwhelmed with planning a wedding," said CEO Tina Loyd. "Hosting the Bridal Fair gives us an opportunity to simplify the planning process for our bridal customers as they prepare for the most important day of their lives."
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Located in Dallas at the southwest corner of Preston and 635, the flagship Terry Costa location has been dressing the women of Dallas-Fort Worth for 26 years. Open 7 days a week, Terry Costa carries everything from prom dresses to Mother of the Bride gowns and more.
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Multi-tonal,patterned pullover sweater. Oversized, slouchy fit. Two vertical seams running down front and back. Small slit at bottom left of front. Contrasting ribbed sleeves.
*98% Cotton, 1% Nylon, 1% Spandex
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Rated 5 out of 5Â by kayach Bring on the Cold! I am officially looking forward to fall weather because of this sweater! It's a bit loose and super soft - the rust/off white combination is also gorgeous! I'm 5'6, 130 and bought a m/l - it was very loose compared to my typical size M, so size down if you are in between. 9.16.2013
Rated 5 out of 5Â by tcdougherty5 Great sweater This sweater looks much prettier in person. Perfect for fall with boots. It is definitely an oversized sweater with substantial weight. It is a must for my fall wardrobe. I am 5'6" tall 130lbs 34c and the xs/s fits perfect. I got the grey. 6.21.2013
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University of Vermont
Admission is FREE and open to the public!
Campus map for location/directions: http://www. uvm. edu/map/
Admission is FREE and open to everyone, including Vermont community members and Deaf community members! The events for Deaf Awareness Week at UVM will occur during the week of September 22-27.
Short-list of Events and Campus Locations:
- Keynote Speaker: Meredith Burke "Ableism in the Deaf Community: Through the Eyes of a Deafdisabled" on Monday, September 22 at 7:00 PM in Davis Center – Grand Maple Ballroom
- ASL Skit Night on Tuesday, September 23, 7:00 PM in Davis Center – Brennan's Pub
- ASL Game Night on Wednesday, September 24, 7:00 PM in Waterman – Memorial Lounge
- ASL Film: "No Ordinary Hero: The SuperDeafy Movie" on Thursday, September 25 at 7:00 PM in Davis Center– Livak Ballroom
- ASL Brunch & Roundtable Discussion on Friday, September 26 10:00 AM in Davis Center – Jost Room
- "Save Austin"e Rally on Saturday, September 27 at 12:00 PM at State House, Montpelier (***disclaimer: UVM will not provide interpreters at this event but there will be volunteer interpreters in attendance)
ASL-English Interpreters will be provided at all of the events. If you are seeking additional accommodations, including, but not limited to Deaf-Blind interpreters or English real-time captioning, please email Hannah at ACCESS: firstname.lastname@example.com no later than September 19th. This deadline will ensure accommodation requests are met. We can accept requests after the date indicated and we will do our best to meet those needs based on availability of services.
Archives of Past Deaf Awareness Week events:
Last modified September 24 2014 03:09 PM


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Dave Spencer and the Big Green Egg Live at Green River This Thursday
"I do not like green eggs and ham. I do not like them, Sam-I-am." - Dr. Seuss
Yeah, well Sam will change his mind when he gets a load of the Big Green Egg at Green River Appliance. Join me this Thursday afternoon from 2pm until 7pm for a live broadcast from Green River Appliance & Electronics at 14th and Daviess in Owensboro.


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Posted by: Marita on: September 5, 2011
5 Years Old this week.
To celebrate I've got 5 days worth of giveaways.
I just want to say a big thank you to everyone who has visited, read, left a comment on the blog, facebook or twitter. Thank you.
Each giveaway is carefully chosen to represent different themes on Stuff With Thing over the last 5 years.
One theme relates to Crohn's Disease – which I was diagnosed with at 19yo. I've been lucky for the last 8 years, my Crohn's has mostly been in remission following a small bowel resection a few months after Annie's birth. However there are times when things flare up and those times are becoming more frequent. A reminder that there is no cure for Crohn's Disease and this is a lifelong condition.
My most popular blog post ever is about poo – "How to Collect a Stool Sample" – now with over 11,300 individual page views!
My very first every blog post was about toilets.
I have found when it comes to living with chronic bowel disease that good quality toilet paper products are vital. Anyone who has ever experienced gastro / diarrhoea will know what I am talking about. When it comes to severe Crohns flareups or pre-Colonoscopy prep the flushable wipes are a godsend.
Which leads me to the first of my blog birthday giveaways.
I would like to thank Kleenex Mums for the very generous giveaway donation and for having the most awesome, softest toilet paper ever.
First Prize: one year's supply of Kleenex Cottonelle toilet paper (300rolls)
(RRP $4.29 per 6pack)
winner can pick any Kleenex Cottonelle variant
(white, marine, kids, hypoallergenic, long roll).
Second Prize: 16 packs of Kleenex Flushable Wipes (RSP $3.98ea)
Third Prize: The cutest Kleenex Plush Puppy ever!
To Enter:
Leave a comment below about when you've needed a gentler toilet paper.
Additional Entries:
Tweet: "Win One Year's Supply of Kleenex Cottonelle Toilet Paper to Celebrate 5 Years of Stuff With Thing: http://www.stuffwiththing.com/?p=7067 at @StuffWithThing"
And leave a comment here saying you Tweeted.
Share on Facebook: "Win One Year's Supply of Kleenex Cottonelle Toilet Paper to Celebrate 5 Years of Stuff With Thing: http://www.stuffwiththing.com/?p=7067 at @Stuff With Thing"
Make sure you @ tag my facebook page so I can verify your entry and leave a comment here saying you shared on facebook.
Competition will close on Friday 30th September 2011 at 6pm AEST.
Competition open to Australian readers only.
Winners will be chosen using Random.org
Disclaimer: Kleenex Mums have donated products for the giveaway only. All opinions expressed are purely my own.
I was a member of The Greens until I tried the recycled toilet paper that they suggested wouldn't destroy the world. My backside still winces at the thought. I have Irritable Bowel Syndrome and making my bowel even grumpier with sandpapering the region is unhelpful.
shared on facebook where all my friends will see it 😉
I am a princess I need it everyday
shared on facebook
Any time that i've eaten the "wrong" foods for my IBS prone tummy :\
I needed one yesterday.
With two kids who seem to spend half their lives in the toilet due to food intolerences, we got through, pardon the french, a shitload of toilet paper!
When we've needed it to be soft and welcoming like Cottonelle ks whenever they've got a tummy upset or gastro bug.
Me? I splashed out on Cottonelle when I was doing my prep for a colonoscopy. Basically you drink a horrible solution over a certain number of hours and spent most of it hovering on the toilet. By bottom said thank you to Cottonelle that day. (Sorry for Overshare, but you did ask…)
Colonoscopys are no fun and "clearing out" your system is a must before the procedure. Having a gentler toiletpaper is always something I make sure I have before my regular checkup!
I tweeted!
I shared on Facebook
I was a no-name recycled toilet paper gal back before children, of course once i had my first (complete with 4th degree tear! ouch) i no longer used the no-name stuff but still went with recycled whenever i could. After hubby had his colonoscopy a new softer tissue was in order and we have tried just about all of them. This is why we need nice and soft tissue in our home always.
Fantastic! This is a prize I could get behind, if you know what I mean! I have to confess to having difficulty with toileting after the birth of both my girls, its just not the same down there and so soft toilet paper and that FABULOUS invention of the moist toilet tissue means I don't have to worry so much anymore! Aaah, the joys of parenting!
I shared on facebook!
As you know we all fell victim to the dreaded gastro very recently. And while the kids stuck to the vomitous variety the grown ups had the great joy of both ends being afftected.
I buy cottonelle anyway, it's our fave. And I was so grateful that week that I do!
So, my gastic system regularly cracks it and if I'm running to the toilet constantly, I am damn well going to have soft toilet paper. Kleenex Hypoallergenic is my loo paper of choice.
To keep my husband happy i brought kleenex and I have to say it was a wise change of brands. It makes sense really as we don't buy sand paper for our nose so why do we do it for our bums?
Switched to Kleenex cottonelle after my first child and massive tears! No turning back since!
Tweeted, and facebooked tooo
Hello Lovely – I had a giggle when I read why you'd chosen this giveaway.
Ironically, I started using cottonelle after I'd had Mr 9lb 12 oz in 2004. The 3rd degree tear he gave me meant that I needed as much "comfort" as I could get…. and the ongoing "fallout" of this particular nether-regional rearrangement has meant I've had to continue to use it.
It is certainly kinder to a sore tush than the recycled sandpaper I was using previously…
So please count me in on this fantastic giveaway.
Our whole family loves the softer paper…. soft on the tooshie and no paper cuts :), so I'd say every day!
The Kleenex puppy is so adorable….!!
The time we all had the dreaded vomity and gastro bug…………..all 5 of us…..
What tooshie doesn't prefer a softer and more gentle toilet paper! Even though I'm sure my children eat it as we go through so much!!
HI !!! I could tell u many toilet stories but I don't want u all to leave this page LOL…anyway my whole body seems to be de-sensitised to everything ….if i even dare to use cheap toilet paper i get very very itchy so I buy Hypoallergenic always and it costs me a fortune so would be wonderful to win so much toilet paper…( i have to tell u something funny – my hubby thinks i have a toilet paper fetish when i go buy it i have to buy about 40 rolls at once and if i get to about 10 that is a worry and i have to duck out to buy more …would never like to run out of toilet paper!!!!) he he he- he thinks it is hilarious i am sooooooo excited by this giveaway!!!…..an my one bad story hope i don't scare u all away…about 6 yrs ago i got Chryptosporidium which is a bacteria bug and u poo every few minutes or so for at least 2 weeks and somehow i caught it ….o it was terrible and sooooooooooo painful – back then i only had cheap and nasty toilet paper and for ppl who have things like this u would know how painful it is to use cheap stuff….I love Kleenex it is my FAV brand of toilet paper!!!! Thanks sooooooo much for this comp! Luv it!
Poor quality toilet paper makes me think of the kleenex ad were the ladyisin the Dr's office and needs a tissue, goes to wipe it with a cheapy and you can hear the sandpaper tear across her nose *shudder*. Imagine that on your sensitive bits owwww
I agree with some of the above posts, I definitely required a softer toilet paper after the birth of my son! Something I never expected to 'need' after a baby!
Honestly?? I only NEED soft toilet paper because I love it and hate scratchy stuff! However, the year after my son was born I had the runs for an entire year. I finally realized it wasn't normal and got tested for Celiacs. Having the right paper was critical for that year and now it's all we use!
Awesome giveaway. Hilarious stories thanks heaps!
I also tweeted you!! @theaussiemommy
20 weeks pregnant. hadn't been taking my metamusil.
Awful H-bomb.
Cottonelle is the best, I agree! As you know I have coeliac disease and when I get glutened… well its not pretty. Thanks goodness for Cottonelle.
and I have tweeted – great giveaway btw!!
After the birth of my son ,the body was in need of everything that is extra soft. Skin was very sensitive and i feltthe need for a gentle toilet paper.
Tweeted the giveaway on twitter.
"Win One Year's Supply of Kleenex Cottonelle Toilet Paper to Celebrate 5 Years of Stuff With Thing: http://www.stuffwiththing.com/?p=7067 at @StuffWithThing"
I follow you on twitter as well.
Yep, I once had a terrible case of food poisoning at a school camp back in primary school! I spent a lot of time in the loo and being a school camp, the toilet paper was generic (one ply, rough, scratchy). Nothing as good as Kleenex Cottonelle!
I can totally relate to your experience. I have IBS and we stock up on Cottonelle when it's on special because it's the only toilet paper that doesn't rip my skin off after frewuent toilet visits. Sorry if TMI
Shared on FB as Chon Ky
I agree with many others about needing softer toilet paper after the birth of my son. It was recommended to me by many Mums, that gave birth in the only hospital available in our area, to take your own toilet paper. Great advice!
I am in total agreement that post-baby delivery is when I have most appreciated being treated kindly down below My best piece of pre-baby advice was to take flushable toilet wipes and soft toilet paper to the hospital – and I now pass on the same message to my pregnant friends.
I buy Kleenex Cottonelle all the time – but really appreciated it after giving birth. No one mentioned to me the pain in going to the toilet. Number 4 is due soon – I'll be taking my own stash to hospital :0)
I have shared on facebook with a tag.
When growing up if we ran out of toilet paper,my dad would rip up pieces of the age newspaper an leave them at the side of the toilet.I would hate mornings when I woke up and saw that!,I also wondered how the toilet took it ( they must been tougher in those days!)
Have shared on facebook (Rae Graham)
I really needed a soft tissue after having my girls and stitches.
I've just tweeted.
I needed a gentler toilet paper when I ate too much of hubby's super hot curry!
Shared on facebook.
I am sure our kids must eat toilet paper, the way we go through it, especially as recently all five of us recently had the dreaded, er, how do I say it delicately, stomach bug?
OMG I swore by those flushable wipes after having my daughter! 3rd degree tears and 2 hours of pushing left me with some nasty tearing and (eek) haemorrhoids and if it hadn't have been for the gentler toilet paper things could have been much more painful! Oh I can still remember the sting!!
Tweet tweet
Shared on Facebook via Daisy, Roo and Two.
My lovely husband dragged me on a 600km road trip a few weeks after giving birth. I wish I had taken a stash of softer toilet paper with me because the good old public toilets in the park cheap paper was very unforgiving!!!
My hubby was diagnosed with Chrohn's too so toilet-paper is a sensitive subject, I am sure you understand.
I have a dodgy tummy too and before I determined ice cream was the worst food for me to eat I'd need lots of toilet paper…! I had no idea there was something called flushable wipes – where have I been living??? So excited to go and get some!
we always had cheap tp growing up but ecer since having my own home and own children i choose soft over sandpaper everytime tp is one thing where soft is always best.
I'm stealing the answer – I'm a princess and need it ever day!! Love it! LOL!
The kiddies are always bring home nasty bugs to share with the rest of us so we always use the good stuff
Have shared on Facebook also
A roll of soft toilet paper was the first thing I packed in my hospital bag when awaiting the arrival of #2. After experiencing the cheap nasty stuff the hospital expect you to use after havign my first bub, I certainly wasn't reliving that experience a 2nd or 3rd time for that matter!!
Shared on FB
Tweeted @hootylishous
considering Boo uses BEDDING as toilet paper, methinks he is on the lookout for something softer.
And I also use it as tissues when we have nasty flu's, cheaper, easier and softer than most tissues.
with a small toliet training child and a then Terminally Ill Husband the bathroom was my 5 mins of sanity but also the one place i seemed to spend the most time helping people . i dont think you can surpass the cost for the quality of this Product . and the flushable wipes are a great all rounder convienace even especially after long road trips and high way pit stops .
Right there with you needing the good stuff for the pre-colonoscopy prep. Lordie but I was thankful myself for those Klennex wet toilet wipes. Love your giveaway, thanks for hosting
The first time ever that I went away for a romantic weekend with my boyfriend (now hubby), we went out to dinner. I got the worst case of food poisoning imaginable! We had to cancel the rest of the trip, and made at least 20 stops to use the toilets on the way home! I was admitted to hospital for two days! It was the most terrible time
Have tweeted (twittername queenofcomps)
Have shared on Facebook (Michelle Vamvas)
After the birth of my son – ouch! Will be needing some more really soon!
I have tweeted!
I have shared on facebook!
Luckily I can survive on the cheapest brand available (most of the time..) but when visitors come, I feel the need to provide them with some comfort – and that's when I need the soft rolls on hand!
With toilet training in this house, I find the softer toilet paper makes things so much easier. Everyone is happy!!
With three little ones we have inevitably succumb to the dreaded gastro virus a couple of times. Definitely a time for a gentler toilet paper!
I could say due to the various intolerances and effects on our bowels, or the obsessive hypersensitive aspie who needs to make sure every last bit of fecal matter is obliterated…but the basic facts are (1) soft paper feels better (2) the toilet is my refuge and sanctuary (even if that means turning a blind eye to the possible supervisory nightmare ensuing once my back is turned, or a deafening bashing on the door)…it's a moment of peace. Almost a whole moment. A second perhaps. Well, at least the paper is soft.
Umm….backpacking trip through Thailand and Cambodia. Street food, spicy food, cheap drinks…need I say more.
I converted to the softness of Cottonelle when I was pregnant with #'2 & 3, you are already uncomfortable & go to the loo a million times a day you definitely need some comfort!
Any time I have been pregnant I needed softer paper for my bottom! Seems that sensitivity is everywhere in pregnancy! Now with 7 people in the house we go through a LOT of toilet paper!!
Definately after having a baby. Woo was that area in need of something soft!
Have tweeted! (as @sconeonamission)
Id love to win because we get what we can afford and MissS is toilet training so with 4 of us using the toilet a years supply would come in handy!
and ive tweeted
Coming from a family of 3 girls + mum we were forever running out of toilet paper, my poor dad forever complaining about how quickly we ran out of toilet paper…As adults, all three sisters now place toilet paper above all else, almost…kind of a survival instinct..one sister now works at a company selling toilet paper, the other is overseas trying her best to "share" "her" toilet paper with a flat mate…myself..I have 2 daughters and am just about to run out for the second time this week. Having a years worth of toilet paper would be a dream come true ;-)…better (almost) than a years supply of chocolate
Sorry, I ramble with no direct answer LOL…I've needed it post surgery (twice for myself) and for 2 little ones (toilet training little bottoms)
High school toilets – just coming into my own bowel issues, no idea what was wrong, but boy did that stuff do a number on me! This was pre-toilet wet wipes too
[…] by Marita on October 1, 2011 // Win One Year's Supply of Kleenex Cottonelle Toilet Paper to Celebrate 5 Years of Stuff With Th… […]
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A man spotted stealing merchandise from a Turlock sports store was arrested on drug and theft charges Wednesday morning.
A Turlock woman was arrested on suspicion of felony child endangerment after Turlock Police officers found her 1-year-old and 5-year-old children wandering near a busy intersection.
Turlock's crime rate climbed by 16 percent in 2014, as the city recorded higher rates of violent crimes and larcenies, according to the Turlock Police Department's annual report.
The Turlock Police Department is seeking the identity of a man who attempted to steal the cash box from a pay at the pump machine at a Turlock gas station.
Will the show go on?
A Turlock man convicted of kidnapping and raping a 19-year-old woman has been granted parole after serving 21 years of a 15 years to life sentence.
A state veterinarian, who was called to a chicken ranch outside of Turlock after authorities found thousands of dead and starving hens, testified Tuesday that the death loss was too significant to have been part of a forced molting process.
A woman reported she was kicked and beaten by an unknown man while at a Turlock bar Sunday night.
The Turlock Police Department is investigating a strong-armed robbery after a man fought with a loss prevention agent after stealing cosmetics from a Walgreens store.
A 10-hour sweep Thursday through Merced County by law enforcement resulted in the arrest of 75 suspected gang members, including those sought for violent crimes like murder and attempted murder.
A preliminary hearing for a father and son accused of a hate crime for beating an African-American woman outside a Turlock bar has been rescheduled for July.
A report of three women stealing clothes at a Turlock store turned into a two-county pursuit that ended with the capture of four suspects, one of whom was a parolee at large.
An employee at the Lowe's store in Turlock was arrested Wednesday after it was discovered she had embezzled thousands of dollars from the company over a span of several months.
Attorneys began unveiling their strategies Wednesday for defending two individuals accused of animal cruelty for abandoning 50,000 hens at an egg farm outside Turlock.
The Turlock Police Department has released new details about the shooting late Monday night that ended the life of a 38-year-old man, including that he was approached by a significant number of people moments before he was gunned down.
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The Turlock Police Department is investigating the death of a 22-year-old man who was fatally shot outside a marijuana grow house.
A Turlock man who tried to avoid a search based on his probation status by running from officers, ended up in custody for drug, weapon and resisting charges.
A man wanted in connection with the killing of a 38-year-old Turlock man was apprehended in Modesto during a traffic stop.
The Merced County Sheriff's Department has arrested three Turlock men in connection with armed robberies at two pharmacies.
A federal grand jury returned a 15-count indictment on Thursday against seven individuals, charging them with conspiracy to commit mail fraud and bank fraud, mail fraud and aiding and abetting, and making false statements to a bank in a mortgage fraud scheme, United States Attorney Benjamin B. Wagner announced.
The Oklahoma police officer accused of taking a 15-year-old girl from the Turlock home of her grandparents has been fired from his job and is facing a kidnapping charge in Stanislaus County.
The rate of vehicle thefts in Stanislaus County saw a decline for a second year, according to the latest report from the National Insurance Crime Bureau.
Page 1 of 1
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FACT CHECK — Did El Paso Taxpayers Get Ripped Off On The Downtown DigiWall? [VIDEO]
Did El Paso taxpayers get gypped on the Downtown DigiWall? We do some fact checking.
Did El Paso taxpayers get gypped on the Downtown DigiWall? We do some fact checking.
To pee or not to pee, that is the question. The answer is, if you need to pee, you won't be able to at the newly renovated San Jacinto Plaza.
Last year, a lot of fun was had at the expense of the "Shawver Park Christmas Lights Display". Former City Rep. Eddie Holguin asked the City at the last minute for funding for a holiday lights display that was so dismal, it had the national news media making fun of it.
Mean Girls and musical chairs – that's what El Paso City Council has been like lately, but it looks like things are going to settle down a bit. City Reps who asked to be moved away from City Rep Lily Limon have decided to go back to their regular chairs, and save taxpayers thousands of dollars in the process.
El Paso City Council wants local construction firms to work on local building projects, so today, they enacted the "Hire El Paso First" policy. If you are a local construction firm, you want to attend a meeting that can help you get more work from the City.
The Lincoln Center was supposed to be demolished on October first, but those plans have been put on hold. The land where the Lincoln Center sits has had a long history with Hispanics in the El Paso area, but instead of trying to save it, maybe it should become an exhibit in the El Paso Museum of History.
It's not secret that I'm a big fan of the ballpark, but I'm not a fan of a proposed parking fee for people who live and work in the neighborhood where the ballpark has been built. It's the Old San Francisco area of downtown El Paso, and a new proposed parking fee will do nothing but pick the pockets of residents there to the tune of up to 200 dollars.
75 days. Tuesday morning will mark 75 days since the switch was thrown on the Shawver lights Christmas display. City Rep. Eddie Holguin proposed the lights display in the Lower Valley park on November 12th, almost 3 months ago. When the lights display was approved, it was with the stipulation that Rep. Holguin would get corporate sponsorships to recover the costs for putting on the light display. It's been 3 months and only $12,050 has been raised - time to show us the money, Mr. Holguin!
There has been a lot of controversy over the Shawver Park lights display that was pushed through El Paso City Council by City Rep. Eddie Holguin. Holguin was warned that it was too late to put on a good display, but he called City Manager Joyce Wilson and City Reps 'grinches' when they said it would be best to wait until next year to put together a good display. Ultimately, he got his lights. Well, the fun is coming to an end, and it's coming to an end on Friday.
Congressman Beto O'Rourke is not being allowed to be a member of the Congressional Hispanic Caucus because he is not Hispanic. The CHC's mission is to "voicing and advancing, through the legislative process, issues affecting Hispanics in the United States". But I guess they don't care about "advancing" the 80% of El Paso Hispanics because we voted in the white guy to represent us. Big ups to you, CHC!
Residents in Central El Paso have successfully beaten back a proposed Walmart Neighborhood Market in their neighborhood. So, why is that a bad thing? It's a bad thing because they missed out on the chance to bring jobs to the area, more shopping choices, and a well landscaped retail site.
City Rep. Eddie Holguin demanded yesterday that Paul Foster and Josh Hunt guarantee the 12 million dollars for their part of the ballpark build. He says he just wants to make sure that if MountainStar goes bankrupt that we wouldn't be stuck holding that bag. So I have a couple of questions for the venerable Mr. Holguin....


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[caption id="attachment_43768" align="alignright" width="330" caption="Photo of Carly Fiorina on @CarlyforCA."][/caption]
This is a tough call, because most of the major candidates for California's top races didn't give the social-media thing much of an effort Tuesday.
Let's rank them, from best to worst:
1. Meg Whitman, Republican candidate for governor. Whitman's team had posted six tweets as of this writing, and she gets credit for giving some love to her supporters, including a video of a news guy interviewing two volunteers.
2. Kamala Harris, Democratic candidate for attorney general. Not only does Harris have a strong-looking picture (as opposed to Jerry Brown's cut-in-half one), she added some urgency with a tweet telling everyone there was only five hours left until the polls close, at 8 p.m.
3. Carly Fiorina, Republican candidate for U.S. Senate. Fiorina was busy compared with other candidates, with seven tweets. And her aggressive tone matches her persona on the campaign trail. This Twitpic of a Fiorina-filled-in ballot, after the question "Have you done this today?" could be seen as confident or crass. Another Twitpic (pictured above) shows her working the phones, which is always good for morale.
4. Jerry Brown, Democratic candidate for governor. Brown gets props for issuing so many thank-yous, but otherwise his feed is quite lame. Last night's shout-out to the Giants was tepid ("Well done @SFGiants"), and he shouldn't have posted this photo via Twitpic. It's blurry and makes him look old and tired.
5. Barbara Boxer, Democratic candidate for U.S. Senate. Among Boxer's three tweets Tuesday was this dud: "STAFF: Know your rights as a voter! http://bit.ly/bQJ386"
6 (tie). Beth Krom, Democratic candidate for U.S. House; and Steve Cooley, Republican candidate for attorney general. Both have only one tweet Tuesday, urging people to get out there and vote. Thanks!
8 (tie). Loretta Sanchez and Van Tran, Democratic and Republican opponents for U.S. House, respectively. Sanchez should come out just ahead because she last tweeted Oct. 28; we last heard from Tran on Twitter an eon ago -- Oct. 27. But Sanchez does the Facebook-automated thing, which breaks off the tweet. Terrible etiquette.
10. John Campbell. No Twitter feed at all. And the rest of his social-media campaign is an utter disaster. At the top of his Facebook page is someone telling him why he's not getting the citizen's vote. Then another commenter complaining about her stolen Beth Krom signs. Yee-ikes.