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On Committees and Motivations
Last year, the Accellera VIP-TSC spent quite a lot of time (I know, because I was there) defining a standard library to establish interoperability between OVM and VMM. All in all, I think this was a useful exercise, although it did highlight some of the shortcomings of treating a standards committee like a software development team. At the end, users were left with a nice standard that enables OVM users to include legacy VMM components in their environment and vice versa.
Beginning late last year, the TSC decided to move forward with the long-term goal of developing a single "universal" verification methodology (deemed "UVM") ostensibly to eliminate the "methodology war" between OVM and VMM that some thought might have actually been solved with the Interoperability kit. The TSC voted back in December to use OVM2.0.3 as the starting point for the UVM, a clear indication that not only was the methodology war over, but that OVM had won. Here's where the questions start.
Shortly after the vote, and before any substantive work had been done in the TSC other than some abstract requirements discussions, Mentor and Cadence released OVM2.1 which included some bug fixes from OVM2.0.3 and provided two new features, callbacks and objections, which just happened to surface in subsequent discussions as two of the more significant requirements for UVM. Since nothing much had been done yet, and 2.1 was already available, I made the suggestion that, instead of locking ourselves into OVM2.0.3, that was already outdated, the TSC should instead use OVM2.1 as the starting point for UVM. I was immediately shouted down for having made an outrageously inappropriate motion. I was told that the TSC had voted on 2.0.3 and that was that. Once callbacks and objections were approved as official requirements, I again suggested that OVM2.1 should be used and was not even allowed to make an official motion to that effect.
Fast-forward to last week (4/7/2010) when the TSC approved using OVM2.1.1 as the starting point for UVM. I'm still waiting for an adequate explanation of why this went from being completely inappropriate to passing overwhelmingly in just a couple of months. Could it be that the initial objections were other than technical? If so, I wonder if the motivation behind them are still there, and it's just that moving to 2.1.1 was such an obviously good idea that those who objected initially realized that they could no longer shout it down.
For the health of the TSC, and my own sanity, I think I'll give my colleagues on the committee the benefit of the doubt.
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Posted April 16th, 2010, by Tom Fitzpatrick
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776 LCGC NORTH AMERICA VOLUME 24 NUMBER 8 AUGUST 2006 www.chromatographyonline.com Simultaneous Determination of Positive and Negative Counterions Using a Hydrophilic Interaction Chromatography Method The objective of this work was to develop a universal high performance liquid chromatography method that is capable of simultaneously retaining and separating both cations and anions within a single chromatographic analysis for the purpose of quantification in pharmaceutical products. A zwitterionic stationary phase operated in the hydrophilic interaction chromatography (HILIC) mode in conjunction with evaporative light scattering detection was investigated for the separation and quantitation of 33 commonly used pharmaceutical counter ions, 12 cations, and 21 anions. Using a single gradient chromatographic analysis, both anions and cations were easily separated from each other in addition the parent pharmaceutical molecules also were separated. The zwitterionic stationary phase utilized in this study offers unique separation capabilities based upon its mixed-mode separation mechanism (that is, electrostatic ion chromatography with the positively and negatively charged functional groups on the stationary phase and HILIC). As a result, a generic screening method was devised that allows for counterion determinations regardless of the pharmaceutical salt that is investigated. The unique retention characteristics of this column were evaluated by varying key mobile phase parameters, such as pH, buffer strength, and organic modifier. After examining the changes in retention, response, and resolution, this universal method was then further evaluated for reproducibility for multiple counterion determinations. For counterion determinations, a typical precision of 2.0% was observed for all counterions and most determinations were within 2.5% of the theoretical salt concentration. Thus, a very rugged screening method was developed capable of separating both anions and cations within a single chromatographic analysis. Counterion determinations were demonstrated for 10 pharmaceutically relevant salts. T he separation and quantitation of passed initial toxicology screening. The counterions in the pharmaceuti- most common pharmaceutical salt forms cal industry is an important are sodium salts of acids and hydrochlo- Donald S. Risley and Brian W. Pack determination. During drug develop- ride salts of amines. Ideally, these salts ment, the selection of the correct salt form would be nonhygroscopic, exhibit Eli Lilly and Company Pharmaceutical early in the development process can pre- solid–state stability, and possess high Product Research and Development, Lilly vent repeating toxicology, biological, and aqueous solubility. However, the most Research Laboratories, Indianapolis, stability studies. As a result, development common salt forms do not always possess Indiana. timeline delays can potentially be pre- the best physicochemical properties and vented. The initiation of the salt selection attributes for development success. In Please direct correspondence to Donald process generally takes place for all ioniz- these cases, a multidisciplinary salt-selec- S. Risley at firstname.lastname@example.com. able compounds that successfully have tion process is necessary to find alternative 778 LCGC NORTH AMERICA VOLUME 24 NUMBER 8 AUGUST 2006 www.chromatographyonline.com 1200.00 Cl- Na+ 80% acetonitrile 1100.00 1000.00 Cl- Na+ 70% acetonitrile 900.00 Cl- Na+ Response (mV) 800.00 60% acetonitrile 700.00 Cl- Na+ 50% acetonitrile 600.00 500.00 Cl- Na+ 40% acetonitrile 400.00 Cl- Na+ 300.00 30% acetonitrile 200.00 Na+ Cl- 100.00 20% acetonitrile 0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 Time (min) Figure 1: These chromatograms were generated at 75 mM ammonium acetate, pH 4.8 acetic acid. The organic content was varied from 20% to 80% acetonitrile. As % organic is increased, retention of both Na and Cl were increased. This is consistent with HILIC. acceptable salt forms. Automated salt suppressor is required to reduce the back- graphic run. For example, a zwitterionic selection systems can be used to screen ground signal. Over the last 30 years, IC compound proceeding through salt selec- numerous counterions in various solvent with conductimetric detection has proven tion can form a basic or acidic salt form. systems, which can result in atypical salt to be a very sensitive detector for both When only milligram quantities of mate- forms. The salt forms that are crystalline cations and anions. However, to perform rial are available, a single method of sepa- from this screen will be scaled up for fur- a cation separation, for example, a cation rating both cations and anions would ther evaluation. At this point, the analyst exchange column with a cation suppressor allow for identity, salt confirmation, and typically evaluates the salt forms using is required to get adequate sensitivity. The stoichiometry within a single chromato- high performance liquid chromatography same is true for anions, but utilizes an graphic run. (HPLC) for counterion identity and stoi- anion exchange column and suppressor. The concept of electrostatic ion chro- chiometry confirmation. The final salt An alternative approach would employ matography (EIC), or zwitterionic ion that proceeds into clinical trials typically strong anion or strong cation exchange chromatography (ZIC) as it was later has desirable properties in relation to sta- columns in conjunction with UV detec- named, with a zwitterionic stationary bility, bioavailability, and is most tion for the determination of organic phase for the separation of ions, was first amenable to conventional formulation acids, or evaporative light scattering detec- proposed by Hu and colleagues in 1993 development. The method of counterion tion (ELSD) for detection of inorganic (3). This separation principal is based determination needs to be precise, accu- salts. Capillary electrophoresis (CE) also upon a zwitterionic stationary phase that rate, and rugged so that it easily can be has been shown to be useful for counte- maintains a fixed positive and negative transferred to other analytical laboratories rion analysis and a method for simultane- charge in close proximity to each other. where the active pharmaceutical ingredi- ous determination of anionic metabolites The separation relies on the ability of the ent is routinely monitored to ensure the based upon CE–mass spectrometry (MS) analyte ions to access both the fixed posi- safety, identity, strength, purity, and qual- has been shown to be specific and selective tive charge, in the case of anions, and the ity of the material. This material ulti- (2). fixed negative charge, in the case of a mately will be made into a drug product In general, all of the previous method- cation. As a result of the proximity of the and consumed by the patient. ologies involve more than one column, charges, the analyte ions will be repulsed Several options exist for counterion more than one mobile phase, and in many and attracted at the same time. Thus, a determinations. The most commonly cases more than one mode of detection to unique and sometimes complicated selec- employed determination utilizes ion- determine both cations and anions. An tivity is obtained. Many mechanistic stud- exchange chromatography (IC), which ideal, and sometimes necessary situation ies have been performed that attempt to was introduced in 1975 (1). In IC, con- would allow for the separation of anions outline the charge interactions on a ductivity detection is typically used and a and cations within a single chromato- molecular level. Hu and Haddad reported www.chromatographyonline.com AUGUST 2006 LCGC NORTH AMERICA VOLUME 24 NUMBER 8 779 600.00 550.00 500.00 450.00 Arginine Response (mV) 400.00 350.00 Malate 300.00 250.00 Succinate Citrate 200.00 Glucuronate Glutarate Fumarate 150.00 Tartrate Maleate 100.00 Tosylate Mandelate Napadisylate Glycolate 50.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00 15.00 16.00 17.00 18.00 Time (min) Figure 2: Overlay chromatogram of common organic ions used for pharmaceutical salt selec- tion. Gradient was the same as Figure 1 with 75 mM ammonium acetate buffer (pH 3.8) concentration. the formation of an electrical double layer mechanism was viewed as more compli- (4,5) to explain retention mechanisms. cated than simple ion exchange. In a sep- Okada and Patil modeled zwitterionic arate evaluation of a carboxybetaine zwit- retention based upon Poisson–Boltzmann terionic stationary phase (12), several theory (6). The formation of a Donnan retention trends were documented. First, membrane combined the previous theo- both the positively and negatively charged ries of Hu (electric double layer) and Patil groups impact the separation of anions, (charged surfaces) to explain both elution whereas cations mainly interact with the order and the effect that mobile composi- negatively charged group. The interaction tion has on retention (7,8). However, of anions with the positively charged there have been few applications reported group is influenced by the cation in the that take full advantage of the separating mobile phase, but mainly follows anion- power of this unique stationary phase. exchange principles. A sulfobetaine-type Many of the early applications have uti- zwitterionic stationary phase, similar to lized pure water as the mobile phase, and that used in this investigation, using water as a result have had difficulty separating as a mobile phase, was evaluated for the both anions and cations. A sulfobetaine separation of multiple anions (13). This stationary phase was reportedly not suc- study indicated that anions with large cessful in the simultaneous separation of hydration energies could not be separated inorganic cations (9). In this study, it was because they have very little retention. noted that the simultaneous repulsion and The experiments conducted here will attraction forces prevented the anions and demonstrate that organic modifier can their countercations from achieving an play a key role in the retention of these ion exchange interaction. Thus, the anion molecules based upon the facilitation of is coeluted with its cation. In a later inves- hydrophilic interaction chromatography tigation of a slightly modified zwitterionic (HILIC). Jonsson and Appelblad demon- system (that is, different carbon chain strated the separation of polar and length between charges), simultaneous hydrophilic compounds with a sulfobe- separation of cations and anions was suc- taine-type zwitterionic stationary. This cessfully performed (10). Again, an aque- work focused on the selectivity from a ous eluent with perchlorate–perchloric HILIC perspective, where the effect of acid modifier was chosen because it pro- acetonitrile and methanol was evaluated vided the best separation. for the retention of RNA–DNA bases in Recently, a carboxybetaine zwitterionic an ammonium formate buffer system column was evaluated for the analysis of (14). nutrients in seawater (11). In addition, The approach presented here also uses a the effect of electrolyte concentration zwitterionic column operated in the (KCl) and pH were demonstrated to have hydrophilic interaction chromatography an effect on anion retention. However, the (HILIC) mode with evaporative light Circle 35 780 LCGC NORTH AMERICA VOLUME 24 NUMBER 8 AUGUST 2006 www.chromatographyonline.com Table I: Retention time and peak tailing are noted as a function of pH at constant buffer strength. As pH increases, retention time of cations increases and retention time of anions decreases. Ion pH 3.1 pH 4.5 pH 6.3 Retention Peak Retention Peak Retention Peak Time Tailing Time Tailing Time Tailing Cations ( 1) Sodium 12.3 1.2 13.6 1.1 15.2 1.1 Potassium 12.2 1.3 13.8 1.3 15.7 1.3 Lysine 15.3 1.2 16.2 1.2 19.8 1.3 Diethanolamine 11.2 1.1 12.6 1.1 14.7 1.3 Trizma 12.1 1.1 12.8 1.1 13.2 1.1 Piperazine 12.3 * 12.9 1.7 12.6 1.2 Choline 10.8 * 12.3 1.1 14.2 2.5 Anions ( 1) Chloride 10.5 1.1 9.9 0.8 9.7 0.9 Bromide 9.9 1.0 8.6 0.7 8.5 1.0 Nitrate 7.0 0.7 5.8 0.6 5.9 0.7 Esylate 7.7 0.7 6.4 0.7 6.5 0.8 Mesylate 8.9 0.8 7.8 0.7 7.8 0.8 Isethionate 9.6 1.0 8.7 1.1 8.8 1.0 Edisylate 13.2 1.2 12.0 1.0 11.6 1.0 Anions ( 2) Sulfate 14.3 1.4 12.9 1.0 12.4 0.9 Cation ( 2) Zinc 13.4 1.6 23.0 2.3 † † Magnesium 17.6 1.6 19.7 2.0 † † Calcium 18.2 2.0 20.5 2.3 † † Anions ( 3) Phosphate 13.1 1.4 13.3 1.7 13.2 1.6 *Not baseline resolved. Calculation of tailing not performed. †Were not eluted under these conditions. scattering detection (ELSD). The combi- that of the reversed-phase mode. drates (29–31), synthetic polymers (32), nation of separation mechanisms (that is, Although the HILIC mode is more simi- steroids (33), and amino acids (34,35). HILIC and EIC) can, theoretically, com- lar to the normal phase and polar organic The HPLC–ELSD system also has been plicate the understanding of the separa- modes, it is different in that the HILIC extremely useful for the determination of tion mechanism; however, the utility of mobile phases contain a relatively high pharmaceutical impurities, raw materials, the zwitterionic column is greatly amount of water (typically 5–40%) as the cleaning verification and small organic enhanced with the addition of organic to strong eluent, which can provide a signif- compounds (36–39). A more recent niche the mobile phase to take advantage of the icant solubility advantage for very for ELSD in the pharmaceutical industry HILIC effect. Alpert first coined the term hydrophilic samples. The HILIC mode is for the detection and quantitation of hydrophilic interaction chromatography can be generated by a variety of polar sta- counterions from pharmaceutical salt for the separation of proteins, peptides, tionary phases. Examples are piperazine forms. Our laboratory first introduced the and polar molecules (15), although this which has been determined utilizing the applicability of HPLC–ELSD for the mechanism had been previously estab- HILIC mode on a cyano column (18) and detection and quantitation of inorganic lished for the separation of carbohydrates polar pharmaceutical analytes which have ions, such as chloride and sodium (16,17). The HILIC mode employs polar been separated using both amino and sil- (40–42). A comparison of the stationary phases with mixed ica columns (19). The HILIC mode also HPLC–ELSD technique with ion chro- aqueous–organic mobile phases creating a has been employed for chiral separations matography, capillary electrophoresis, and stagnant enriched water layer around the using cyclodextrin and macrocyclic titration for the determination of Cl in polar stationary phase. This enriched layer antibiotic based packings (20,21). pharmaceutical drug substances has been allows analytes to partition between the In HPLC, ELSD has an extensive compared statistically and it was deter- two phases based upon their polarity. In application base, but it is especially mined that the four techniques were contrast to reversed-phase chromatogra- important when UV detection is not fea- equivalent (41). However, ELSD is a cost phy, where a nonpolar stationary phase is sible. The concept and operation of com- effective method that can be used with employed and analyte elution is facilitated mercially available evaporative light-scat- many other HPLC applications in addi- by the organic strength of the mobile tering detectors as sensitive and universal tion to the analysis of counterions (for phase, analyte elution is facilitated by the has been discussed thoroughly in the liter- example, assay and impurity determina- aqueous (more polar) component of the ature (22). ELSD has been shown to suc- tions for compounds lacking a strong mobile phase in HILIC mode. The sepa- cessfully detect many substances, such as chromophore) which gives it a unique ration mechanism and retention order in phospholipids (23–26), triglycerides, fats advantage over other techniques. the HILIC mode is therefore opposite to and fatty acid esters (27,28), carbohy- The goal of this article is to show the www.chromatographyonline.com AUGUST 2006 LCGC NORTH AMERICA VOLUME 24 NUMBER 8 781 Table II: Effect of pH on the retention and peak shape of organic ions. The effect is not quite as significant as with inorganic ions. Same chromatographic conditions as outlined previously were utilized. Ion pH 3.8 pH 4.8 pH 6.0 Retention Peak Retention Peak Retention Peak Time Tailing Time Tailing Time Tailing Anions ( 1) Citrate 12.5 2.2 13.3 1.5 13.1 1.4 Glucuronate 13.1 0.8 12.7 0.9 12.5 1.0 Mandelate 5.2 0.8 5.4 0.7 5.4 0.7 Succinate 9.2 1.3 12.5 1.1 12.4 0.9 Tartrate 13.6 1.7 13.2 1.6 12.6 1.4 Fumarate 12.7 1.3 13.0 1.2 12.3 1.1 Glycolate 10.6 0.9 10.9 0.9 10.8 0.8 Glutarate 8.9 1.4 12.7 1.1 12.5 1.0 Maleate 4.4 0.9 4.8 0.8 6.3 0.9 Malate 11.9 1.3 12.9 1.3 12.6 1.0 Tosylate 3.8 1.0 3.5 0.9 3.6 1.0 Napadisylate 13.8 1.2 12.4 1.1 11.9 1.1 Cations ( 1) Benzylamine 10.9 1.1 10.5 1.1 10.2 1.2 Arginine 14.7 1.2 15.2 1.3 17.7 1.4 *Notbase line resolved. Calculation of tailing not performed. **Did not elute under these conditions. application of a relatively new column tonitrile–15% buffer and mobile phase B Results and Discussion technology operated in the HILIC mode, was 10% acetonitrile–90% buffer. The Effect of Organic Composition: The first while fully taking advantage of the EIC buffer comprised ammonium acetate and important aspect of this work was to interaction, for the simultaneous separa- pH adjusted with acetic acid (buffer con- establish whether the zwitterionic station- tion and quantitation of cations and centration and pH were varied and noted ary could exhibit a HILIC effect for the anions within a single chromatographic in the text). An Orion model 720A pH separation of inorganic cations and run with ELSD as a universal detection meter from Orion Research, Inc. was used system. to measure the pH of the mobile phase buffers (Beverly, Massachusetts). The gra- Experimental dient system employed with each injec- Chemicals: Acetonitrile was purchased tion was as follows: 0–2 min at 100% A, from Burdick and Jackson (Muskegon, 2–22 min a linear gradient to 100% B, Michigan). A sodium and chloride stan- 22–25 min at 100% B, 25–26 min a lin- dard solution was acquired from Fluka ear gradient back to 100% A, and equili- Chemika (Buchs, Switzerland). The pH brate 26–35 min at 100% A. Note that buffers were from Red Bird Service this gradient is opposite of conventional (Osgood, Indiana). Deionized water and reversed-phase HPLC due to the fact that nitrogen were from an in-house system. HILIC is employed. All other chemicals were obtained from Standard and sample preparation: Sigma-Aldrich Chemical Company (St. Three individual standards were weighed Louis, Missouri). accurately and diluted with mobile phase Equipment: The HPLC system con- A or accurately pipetted from a standard sisted of a Hewlett Packard 1050 pump stock solution and diluted with mobile and auto sampler (Wilmington, phase A. The standard curve for three cal- Delaware) integrated with an Alltech 800 ibration standards was calculated by least- evaporative light scattering detector from squares regression analysis of peak area Alltech Associates (Deerfield, Illinois). versus concentration. The samples were The detector was operated at 55 °C, 3.5 weighed individually and the weights bar nitrogen and a gain setting of 1 were based upon the theoretical content throughout the experiments. A ZIC- of the counterion to be within the stan- HILIC column (250 4.6 mm, 5 m) dard range. The concentration of the from SeQuant was used for the separation counterion in the samples was determined (Umea, Sweden). The mobile phase flow by comparing the peak area to the stan- rate was set at 1.0 mL/min and injection dard curve. volumes of 10 or 20 L were used. Mobile phase A make-up was 85% ace- Circle 36 782 LCGC NORTH AMERICA VOLUME 24 NUMBER 8 AUGUST 2006 www.chromatographyonline.com Table III: Retention time and peak tailing are noted as a function of buffer concentration at a constant pH. As buffer concentration increases, retention time of cations decreases and retention time of anions increases. The best peak shape for both cations and anions was achieved between 50 mM and 100 mM acetate buffer. Chromatography conditions are the same as outlined earlier. Ion 10 mM 25 mM 50 mM 100 mM 150 mM 200 mM Ret. Peak Ret. Peak Ret. Peak Ret. Peak Ret. Peak Ret. Peak Time Tailing Time Tailing Time Tailing Time Tailing Time Tailing Time Tailing Cations ( 1) Sodium 18.5 1.3 15.1 1.3 13.4 1.2 12.3 1.1 12.2 1.1 12.1 0.7 Potassium 19.1 1.6 15.5 1.4 13.7 1.4 12.5 1.2 12.1 1.2 12.1 0.8 Lysine 21.6 1.5 18.6 1.3 16.5 1.2 15.3 1.3 14.9 1.2 14.8 1.2 Diethanolamine 17.6 1.1 14.7 1.3 12.6 1.1 11.5 1.1 11.4 1.1 11.4 0.8 Trizma 17.4 1.0 14.3 1.1 12.7 1.1 11.8 1.1 11.7 1.1 11.7 0.7 Piperazine * * * * 18.5 2.0 16.9 2.1 16.4 1.8 14.1 0.8 Choline 17.3 1.3 13.8 1.2 12.0 1.1 11.1 1.1 11.0 † 10.9 † Anions ( 1) Chloride 7.7 0.5 9.1 0.6 10.1 0.9 10.5 1.0 10.8 1.1 11.0 1.0 Bromide 5.8 0.6 7.5 0.6 8.8 0.7 9.8 1.0 10.1 1.1 10.4 0.9 Nitrate 4.2 0.6 5.1 0.6 6.0 0.6 7.1 0.7 7.6 0.7 8.1 0.8 Esylate 4.3 0.6 5.4 0.6 6.5 0.6 8.1 0.8 8.9 0.9 9.5 1.1 Mesylate 5.3 0.6 6.7 0.6 7.9 0.7 9.3 0.9 9.8 1.0 10.1 1.2 Isethionate 5.6 0.6 7.6 0.6 8.9 0.8 10.0 1.1 10.4 1.1 10.6 0.8 Edisylate 9.7 0.6 11.7 1.0 12.1 1.1 12.6 1.2 12.9 1.2 13.1 1.2 Citrate 10.5 2.0 12.6 2.5 13.2 2.5 13.6 2.8 13.8 2.7 14.1 0.8 Anions ( 2) Sulfate 11.5 0.7 12.7 0.9 13.3 1.0 13.9 1.1 13.7 1.2 13.9 1.1 Cations ( 2) Zinc 29.0 2.0 23.5 2.0 22.4 2.2 20.3 2.0 20.4 2.4 19.8 2.2 Magnesium 29.0 2.0 22.5 2.0 19.3 1.7 16.3 1.4 15.2 1.3 14.2 1.2 Calcium 29.3 2.0 23.3 2.6 20.1 2.2 17.0 1.7 15.8 1.5 15.1 1.4 Anions ( 3) Phosphate 11.4 0.8 13.0 1.6 13.5 1.8 13.6 1.9 13.7 1.6 13.9 1.4 *Was not eluted under these conditions. †Notbase line resolved. Calculation of tailing not performed. anions. A concern was that the electro- tion of anions and cations in the HILIC observed. These experiments were con- static effect would dominate the separa- mode. For all of the investigation in this ducted with constant buffer strength of tion and the organic modifier would have work, acetonitrile was used because it 50 mM ammonium acetate. The mobile very little effect on the selectivity. As can already has been demonstrated that ace- phase starting point in this case was 85% be seen in the bottom chromatographic tonitrile will promote the HILIC effect acetonitrile–15% buffer (pH was adjusted trace in Figure 1, Na and Cl are not more so than methanol. In addition, Fig- with acetic acid) and a flow rate of 1 separated with 20% acetonitrile–80% ure 1 also illustrates that even for a simple mL/min was used (see equipment section buffer. As the organic content of the separation of Na and Cl ; the run time for gradient). mobile phase is increased from 20% to can become excessively long. Thus, a gra- A measurable effect on retention is 80% acetonitrile, the retention times of dient (that is, opposite of a typical observed as the pH is increased from 3.1 both ions are increased substantially from reversed-phase separation) will be used for to 6.6 for both cations and anions. Inter- 3.5 min to 10.5 min for the chloride ion all future separations with the under- estingly, as the pH was increased, the and to approximately 20 min for the standing that for any compound– retention times of all of the cations sodium ion. In addition, the resolution counterion separation, the run time could increased and the retention times of the between the ions increases with increased be optimized for an isocratic separation. anions decreased (see Table I). The change organic composition. In a typical The data generated will be a gradient in retention was most drastic for the 2 reversed-phase interaction (not that group ramped from 15% aqueous buffer to 90% cations where calcium, magnesium, and I cations are retained typically on a aqueous buffer. zinc were not eluted under these gradient reversed-phase column), these ions would Effect of pH: pH effects were evaluated conditions at pH 6.6. The effect on cation be eluted in the solvent front for all across a range of approximately 3.1–6.6. retention is presumably due to the H mobile phase compositions. In the same Across this range the sulfobetaine-type interacting with the negatively charged fashion, in a completely aqueous system zwitterionic stationary phase retains its part of the zwitterions (SO3 ), which with a zwitterionic stationary phase, these permanent positive and negative charges. ultimately shields the cation from having ions would have been coeluted, which had Because there is no change in ionization a strong interaction at a lower pH. The been reported previously as an ion-pairing state of the analyte ions (for the inorganic anions are following standard ion effect (43). Therefore, this is a strong indi- ions) or stationary phase across the pH exchange theory. As can be seen in Table I, cator that organic composition of the range, it was presumed that pH differ- there is a minimal effect of pH on peak mobile phase is an extremely powerful ences would have a minimal effect on ion shape except for the 2 ions. In this case, tool in controlling selectivity and reten- retention. However, a definite trend was a lower pH is recommended to ensure www.chromatographyonline.com AUGUST 2006 LCGC NORTH AMERICA VOLUME 24 NUMBER 8 783 Table IV: Counterion determinations for 10 pharmaceutical salts Sample Counterion Counterion Counterion % of RSD % Drug R2 Result by HPLC Theory % Theory (n 3) Retention time (min) Trazodone HCl Chloride 8.70 8.68 100.2 1.31 3.76 Ranitidine HCl Chloride 9.85 10.10 97.5 1.28 8.29 Imipramine HCl Chloride 11.06 11.19 98.8 1.30 5.33 0.9999 Verapamil Hcl Chloride 7.36 7.22 101.9 1.45 4.19 Chloropromazine Chloride 9.83 9.98 98.5 0.45 5.23 HCl Proglumide Na Sodium 6.19 6.45 96.0 1.28 3.31 0.9998 Antazoline Phosphate 23.30 26.13 89.2 1.03 5.10 0.9918 phosphate Pantothenic Calcium 8.43 8.38 100.6 0.26 10.50 0.9999 acid Ca Enapril maleate Maleate 24.15 23.57 102.5 1.39 3.86 0.9992 Fenoterol HBr Bromide 20.89 20.79 100.5 2.03 8.26 0.9986 For organic ions, the trends (see Table II) are not as clearly defined due to the (a) pKa of the acids. Dependent upon the ionization state of the ion, the retention mechanism could change from ion 400.00 exchange to one that is affected by hydro- gen bonding. However, a reasonable sepa- e ration is obtained for multiple (14) Response (mV) 300.00 organic ions that are commonly used to synthesize pharmaceutical salts (Figure 2). Figure 3 represents the separation of 200.00 mainly the inorganic ions that were evalu- ated within this work (see Table I). Inter- 100.00 estingly, ions of a particular charge state are eluted within distinct regions of the chromatogram under these separation 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 16.0 17.0 18.0 19.0 20.0 21.0 22.0 conditions. For example, Figure 3 demon- Time (min) strates that the elution order is, in general (b) 1 1 2 3 2. The 2 and PO4-3 3 anion elution order is predicted only 240.00 Cl- SO4-2 +2 Ions from SO4 2 and PO4 3 and is not as reli- 220.00 Na+ K+ able as the predictions that do not include 200.00 polyatomic ions (for example, lysine Br- 180.00 elutes after sulfate and phosphate). How- Response (mV) 160.00 Mg2+ Ca2+ ever, this elution order is a powerful tool 140.00 in understanding the interactions that 120.00 dominate the separation. For example, all 100.00 1 ions are eluted before all 1 ions, 80.00 which indicates that the fixed SO3 func- 60.00 Zn2+ tionality on the stationary phase has a 40.00 strong interaction with cations because it 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 16.0 17.0 18.0 19.0 20.0 21.0 22.0 is more accessible. Time (min) Effect of buffer concentration: Because ELSD was used in this investiga- Figure 3: Chromatograms collected at 100 mM ammonium acetate buffer. Gradient from 85% acetonitrile to 10% acetonitrile in 20 min. Regions where ions typically are eluted under tion, a volatile buffer of some sort was these conditions are indicated. required for the detection of the ions. For example, ammonium acetate buffer can that the ions will be eluted and better two cations essentially are coeluted and at be used in the mobile phase so that a par- peak shape will be obtained. The separa- pH 6.6 they are slightly separated with a ticle of Na CH3COO will be formed tion of Na and K is fairly difficult retention time difference of approxi- during desolvation in the detector drift under these conditions. At pH 3.1, these mately 30 s. tube and subsequently detected by light 784 LCGC NORTH AMERICA VOLUME 24 NUMBER 8 AUGUST 2006 www.chromatographyonline.com concentration was increased to 100 mM, the peaks symmetry improved, however, there were no improvements beyond that 1200.00 point. At 200 mM, the 1 cations actu- 1100.00 ally began to exhibit significant peak 1000.00 fronting. From this experiment, a range of 900.00 50 mM–100 mM buffer concentration 800.00 was a recommendation for these experi- ments. Phosphate buffer was considered Response (mV) 700.00 600.00 so as to allow for UV detection of the 500.00 organic acids, however, the solubility in 400.00 high organic is limited and also dimin- 300.00 ishes further as the pH of the aqueous 200.00 portion increases (when mixed with ace- 100.00 tonitrile). 0.00 The retention of all ions investigated, 0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00 Time (min) in most cases beyond 4 min, is very con- venient for counterion determinations. When operating in HILIC mode nonpo- Figure 4: Overlay chromatograms of several pharmaceutical salts. In this example, the coun- terions are easily separated from each other. The molecules are separated from each other as lar compounds will be eluted with very well. Chromatography conditions are using 75 mM ammonium acetate buffer (pH 5.0) and little retention because they are portioned the same gradient as described earlier. Salt forms are identified and quantitated in Table III. preferentially into the organic layer. Even though these compounds will be charged in many cases, the organic content will scattering. In the case of Cl , under these mechanism. dominate the retention mechanism and same conditions, NH4 Cl is formed Because the main mechanism of inter- the compound should be eluted before and subsequently detected. In this study, action in the HILIC mode is based upon counterions. ammonium formate and ammonium a partitioning of the ions into the aqueous Counterion determination: This work acetate were evaluated. Ammonium for- phase that forms a stagnant layer on the was concluded by evaluating the zwitteri- mate offered no advantages over ammo- stationary phase surface, the decrease in onic column operated in the HILIC nium acetate. Therefore, an ammonium retention time might be best understood mode, in conjunction with ELSD, by acetate–acetonitrile system was evaluated by a shift in equilibrium concentrations. determining the counterion concentra- for all experiments. In addition to allow- As the NH4 concentration increases tion in 10 pharmaceutically relevant salts. ing for the detection of ions by ELSD, the preferentially in the aqueous layer, there is trazodone HCl, ranitidine HCl, buffer concentration has a pronounced less opportunity for the analyte counteri- imipramine HCl, verapamil HCl, and effect on the chromatography, and in ons to partition into the aqueous layer chlorpromazine HCl were chosen as the combination with organic concentration (44). Thus, the ions are swept through the representative hydrochloride salts. Proglu- appears to be the most important variable column (mainly in the organic layer) with mide Na, antazoline phosphate, pan- in controlling selectivity. As can be seen in less interaction with the column and the tothenic acid Ca, fenoterol HBr, and Table III, when the buffer concentration aqueous phase. In addition, as the NH4 enapril maleate also were evaluated. A gra- is increased from 10 mM to 200 mM interacts strongly with the SO3 fixed dient was again employed to demonstrate ammonium acetate, both the peak shape negative charges as the buffer concentra- the resolving power and the utility of a and retention times of the ions are drasti- tion increases, access to these fixed charges universal method for separation of a cally affected. This experiment was run is diminished. As a result, cations do not counterion from the parent molecule. A with a gradient from 85% acetoni- interact with SO3 and are not signifi- starting mobile phase of 85% acetoni- trile–15% ammonium acetate to 10% cantly retained; anion retention is affected trile–15% 75mM ammonium acetate acetonitrile–90% ammonium acetate at in the opposite manner. The anions do (pH 4.8 with acetic acid) with a 2-min approximately pH 5 at a flow rate of 1 not experience the typical repulsion forces hold, and gradient to 90% aqueous buffer mL/min (gradient described in experi- of the SO3 functionality and can then were chosen based upon previous data mental section). access the tertiary amine for ion exchange. (see experimental section for gradient). As expected, and reported previously, This ion exchange interaction causes the The linearity of standards was first evalu- the buffer concentration has a significant anions to be retained more strongly. ated. Excellent linearity (typical R2 of impact on retention and peak shape of In addition to retention time effects, 0.999) of a three point standard was ions. The trend observed while increasing buffer concentration also impacts peak observed for all ions that were quanti- buffer concentration from 10 mM to 200 shape. With a very low buffer concentra- tated. The same calibration curve was uti- mM was that cations were not retained as tion (10 mM), the peak shapes exhibited lized for all of the HCl salts. Standards long, and anion retention increased. severe fronting for the anions and in most were typically prepared in the range of Again, this can be explained by a two-part cases tailing for the cations. As the buffer 0.2–0.7 mg/mL of the counterion, while www.chromatographyonline.com AUGUST 2006 LCGC NORTH AMERICA VOLUME 24 NUMBER 8 785 the samples were prepared in a concentra- excellent agreement with theory for all of (22) A. Stolyhwo, H. Colin, and G. Guiochon, J. tion to fall within the standard range. As the counterion determinations with most Chromatogr. 265(1), 1–18 (1983). can be seen from Figure 4, the com- values within 2.5% of the theoretical salt (23) W.S. Letter, J. Liq. Chromatogr. 15(2), pounds are separated from each other concentration. In summation, a universal 253–266 (1992). under these conditions as well as all of the set of HPLC conditions with one col- (24) J.S. Perona and V. Ruiz-Gutierrez, J. Sep. Sci. counterions. Again, this demonstrates the umn, one mobile phase, and one detec- 27(9), 653–659 (2004). power of the gradient ZIC-HILIC effect tion system, was developed that would (25) S.L. Abidi and T.L. Mounts, J. Chromatogr. A as a universal screening method. The suffice for the determination of a large 773(1–2), 93–101 (1997). counterion-determination data are sum- population of pharmaceutically relevant (26) F. Mancini, E. Miniati, and L. Montanari, marized in Table IV for multiple salts. The salts. Italian J. Food Sci. 9(4), 323–336 (1997). RSD for all measurements was less than (27) A. Stolyhwo, M. Martin, and G. Guiochon, 2.0% for three replicates and the maxi- References J. Liq. Chromatogr. 10(6), 1237–1253 mum absolute difference between the the- (1) H. Small, T.S. Stevens, and W. Bauman, (1987). oretical salt concentration and the experi- Anal. Chem. 47(11), 1801–1809, (1975). (28) T. Andersen, A. Holm, I. L. Skuland, R. mentally determined value was 2.8% for (2) T. Soga, Y. Ueno, H. Naraoka, Y. Ohashi, M. Trones, and T. Greibrokk, J. Sep. Sci. antazoline phosphate. In the case of the Tomita, and T. Nishioka, Anal. Chem. 26(12–13), 1133–1140 (2003). phosphate salt, which exhibited the largest 74(10), 2233–2239 (2002). (29) R. Macrae, L.C. Trugo, and J. Dick, Chro- error from theory, the difference could be (3) W. Hu, T. Takeuchi, and H. Haraguchi, matographia 15(7), 476–478 (1982). attributed to the low solubility of phos- Anal. Chem. 65(17), 2204–2208 (1993). (30) B.A. Kimball, W.M. Arjo, and J.J. Johnston, phate in high organic concentrations, (4) W. Hu, Langmuir 15(21), 7168–7171 J. Liq. Chromatogr. Rel.Technol. 27(12), although this sample was not investigated (1999). 1835–1848 (2004). further. However, most errors were within (5) W. Hu and P.R. Haddad, Trends in Anal. (31) Y. Wei and M.Y. Ding, J. Chromatogr. A, 0.3% absolute, which is consistent to pre- Chem. 17(2), 73–79 (1998). 904(1), 113–117 (2000). viously reported quantitative data (41). (6) T. Okada and J. M. Patil, Langmuir 14(21), (32) P.D. Green, H. Meng, and J.E. Seely, Polymer 6241–6248, (1998). Preprints (American Chemical Society, Divi- Conclusion (7) H.A. Cook, W. Hu, J.S. Fritz, and P.R. Had- sion of Polymer Chemistry) 38(1), 608–609 This work demonstrates the separating dad, Anal. Chem. 73, 3022–3027 (2001). (1997). power of the zwitterionic stationary phase (8) H.A. Cook, G. Dicinoski, and P.R. Haddad, (33) P.A. Asmus and J.B. Landis, J. Chromatogr. for the simultaneous retention and separa- J. Chromatogr., A 997(1-2), 13–20 (2003). 316, 461–472 (1984). tion of cations and anions, especially (9) W. Hu and H. Haraguchi, Anal. Chem. (34) J.A. Peterson, L.J. Lorenz, D.S. Risley, and when organic eluent is used in the mobile 66(5), 765–767 (1994). B.J. Sandmann, J. Liq. Chromatogr. Rel. Tech- phase. The column was successfully (10) W. Jiang and K. Irgum, Anal. Chem. 71(2), nol. 22(7), 1009–1025 (1999). demonstrated to operate in the HILIC 333–344 (1999). (35) H.J. C. Das Neves, and Z.B. Morais, J. High mode as a retention mechanism, where (11) C.O. Riordain, P. Nesterenko, and B. Paul, J. Resolut. Chromatogr. 20(2), 115–118 (1997). the retention times of Cl and Na were Chromatogr. A 1070(1-2), 71–78 (2005). (36) D.S. Risley and J.A. Peterson, J. Liq. Chro- increased from 3.5 min to 10.5 min and (12) W. Hu, P.R. Haddad, K. Tanaka, and K. matogr. 18(15), 3035–3048 (1995). 20 min, respectively. The pH of the Hasebe, Anal. Bioanal. Chem. 375(2), (37) M. Rajevic and P. Betto, J. Liq. Chromatogr. mobile phase had an effect where the 259–263 (2003). Rel. Technol. 21(18), 2821–2830 (1998). retention of the cations was decreased and (13) T. Umemura, S. Kamiya, A. Itoh, K. (38) D.S. Risley, K.F. Hostettler, and J.A. Peter- the retention of anions was increased. Chiba, and H. Haraguchi, Analytica Chimica son, LCGC 16(6), 562–568 (1998). With the exception of the 2 cations, Acta 349(1–3), 231–238 (1997). (39) C.E. Kibbey, Mol. Diversity 1(4), 247–258 which required lower pHs to be eluted, (14) T. Jonsson and P. Appelblad, LCGC (1996). the use of pH is considered more of a 17(Suppl.), 72-73 (2004). (40) J.A. Peterson and D.S. Risley, J. Liq. Chro- means to improve peak shape as opposed (15) A.J. Alpert, J. Chromatogr. 499, 177–196 matogr. 18(2), 331–338 (1995). to control selectivity. Buffer concentra- (1990). (41) D.S. Risley, J.A. Peterson, K.L. Griffiths, and tion, as suspected with EIC interactions, (16) L.A. Th. Verhaar, and B.F.M. Kuster, J. Chro- S. McCarthy, LCGC 14(12), 1040–1047 is the most important parameter (in com- matogr., 234 (1), 57–64 (1982). (1996). bination with organic content) for con- (17) P. Orth and H. Englehardt, Chro- (42) M.D. Lantz, D.S. Risley, and J.A. Peterson, J. trolling selectivity, resolution, and for matographia, 15(2), 91–96 (1982). Liq. Chromatogr. Rel. Technol. 20(9), optimizing run times. These experiments (18) C. McClintic, D.M. Remick, J.A. Peterson, 1409–1422 (1997). clearly indicate that a large number of and D.S. Risley, J. Liq. Chromatogr. Rel. Tech- (43) W. Hu, H Tao, and H. Haraguchi, Anal. cations (12) and anions (21) can be sepa- nol. 26(18), 3093–3104 (2003). Chem. 66(15), 2514–2520 (1994). rated and ultimately detected by ELSD. (19) B.A. Olsen, J. Chromatogr. A 913 (1-2), (44) B.W. Pack and D.S. Risley, J. Chromatogr. A Finally, the gradient was applied to 10 113–122 (2001). 1073(1–2), 269–275 (2005). pharmaceutical salts for the determination (20) S.C. Churms, J. Chromatogr. A 720(1–2), of the counterion. In this experiment, 75–91 (1996). excellent linearity was observed with an (21) D.S. Risley and M.A. Strege, Anal. Chem. R2 0.999 in most cases. There was 72(8), 1736–1739 (2000).
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Extended interview with Wayne Swan
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Reserve Bank Govenor Glenn Stevens says Australia is officially in recession.
Topics: international-financial-crisis, business-economics-and-finance, economic-trends, markets, trade, australia
A man serving six years in jail for stabbing a friend who died almost two years later has faced Port Augusta court in an Aboriginal round table conference.
Topics: courts-and-trials, indigenous-aboriginal-and-torres-strait-islander, law-crime-and-justice, thevenard-5690, sa, port-augusta-5700, port-lincoln-5606, port-pirie-5540
Posted| Updated
The West Australian Police Minister, Rob Johnson, says the Office of Crime Prevention is under review but won't reveal whether any jobs are on the line.
Topics: public-sector, government-and-politics, budget, crime, perth-6000, wa
Posted| Updated
Pro-abortion activists rally in Brisbane to support a 19yo facing jail for allegedly aborting her baby with illegal drugs.
Topics: abortion, health, reproduction-and-contraception, brisbane-4000, australia, qld
Teachers say a $550 million Federal Government program to recruit young teachers is an insult.
Topics: teachers, education, government-and-politics, federal-government, australia
Posted| Updated
Port Adelaide's Matt Thomas has been given a one-game suspension and fined $2,600 after being found guilty of reckless contact with an umpire by the AFL tribunal.
Topics: australian-football-league, port-adelaide-5015, nsw, qld, sa, vic, wa
Posted| Updated
Comedy duo Dawn French and Jennifer Saunders are to receive a British Academy of Film and Television Fellowship at the Academy's 2009 television awards.
Topics: television, arts-and-entertainment, england, united-kingdom
China has long been criticised for shielding rogue regimes like North Korea, Zimbabwe and Burma from tough international action.
Topics: unrest-conflict-and-war, government-and-politics, world-politics, china
A Queensland man imprisoned for child-sex offences has had his bail application rejected in the Supreme Court in Cairns.
Topics: sexual-offences, law-crime-and-justice, courts-and-trials, cairns-4870, qld, papua-new-guinea
A seven-year study of birdlife in the central west of New South Wales has found the bird populations are in decline.
Topics: birds, environment, conservation, endangered-and-protected-species, environmental-management, land-clearing, animal-science, cowra-2794, act, nsw
A 57-year-old man has died in an accident on a property at Yeppoon in central Queensland.
Topics: accidents, accidents---other, disasters-and-accidents, yeppoon-4703, qld
Posted| Updated
The firefighter who ordered sprinklers be turned off in Hobart's Myer store before a fire was under control says it was necessary to protect his crew and find the source of the blaze.
Topics: industrial-fires, courts-and-trials, hobart-7000, tas
A central Queensland seismologist says it is only a matter of time before the region is hit by an earthquake.
Topics: earthquake, research, rockhampton-4700, bundaberg-4670, gladstone-4680, yeppoon-4703
Posted| Updated
The authors of a world-first study say it proves that poor indigenous health is caused by social rather than biological factors.
Topics: indigenous-aboriginal-and-torres-strait-islander, community-and-society, health, child-health-and-behaviour, australia, canada, new-zealand, united-states
Posted| Updated
Carlton Football Club members say their thoughts go out to the family of Richard Pratt.
Topics: community-and-society, person, australian-football-league, melbourne-3000, vic
Posted| Updated
Authorities apologise to those injured in the Morley bus crash.
Topics: road, accidents, disasters-and-accidents, morley-6062, wa, perth-6000
For some people the great Australian dream is home ownership, but according to a new survey it is a dream that many feel is out of reach.
Topics: economic-trends, business-economics-and-finance, industry, housing-industry, housing, australia


text: Listeners Profiles Everyone (2) Profiles matching: not telling you that either!!! Everyone Lendiner Hometown: D.C School: Haha not gonna tell you. Employer: Not telling you that either. Stations: Ed Sheeran Radio, Lorde Radio, A Thousand Years (Part 2) Radio, Pitch Perfect (Film Cast) Radio voyager51 Hometown: Not Telling You School: Not Telling You Employer: Not Telling You That Either Stations: Today's Hip Hop and Pop Hits Radio, Maroon 5 Radio, EDM Festival Dance Radio, He is We Radio


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Fits: 35DN, 102, 112, 212DLX, 1000, SL-61, SL-51S (Japnese Version) S103, S302, Slik 1000, Sport II, U112, U210, U212DLX
Most Liked Positive Review
Does the Job
The post is a piece of a Slik tripod system. If you use multiple cameras to shoot you can very quickly detach and attach a camera to a tripod by having multiple posts, one for each camera you need to mount on the tripod, already attached to your camera. Not really that different from other plate and clamp tripod systems but smaller, easier, and less expensive to do than for many other tripod systems.
Most Liked Negative Review
When you need it there it is
post is one of those products that you need to make another one work. And work it does.
Reviewed by 24 customers
Good: Right piece to replace missing one. My tripod is working again. Neg: Took 2 more days than expected. Picked standard ground shipping by USPS.
anyone with an older slik should have two of these around for diferent bodies or mounting binoculars
I found a slik tripod at a flea market, but it did not have the quick release post,I like the sturdiness of the slik tripod and the tiny size of the release post,the down side is its easy to misplace
I use it to mount my camera to my tripod. It works great.
fits in the tri-pod perfectly
This was purchased for my fathers tripod. This piece was missing and he wasnt sure if it could be fixed. Thanks to Adorama Camera I found exactly what I was looking quickly and affordably. Pops is now happy with his "new" tripod.
Have two bodies and multi lenses, bought 3 to accomadate quick changes
post is one of those products that you need to make another one work. And work it does.
I have had a Slik Universal 212 Deluxe for better than 16 years and find it to be a very stable and reliable tripod. I like the fact that the head can be removed from the top of the column and placed on an attachment point on either one of the legs or the bottom end of the column. This makes it good for getting down low to the ground. As I am 6'2" the column extends to a comfortable height for me to use. The only drawback to it is that at six (6) pounds, it is rather heavy, so carrying it around in the field can be a bit tiring.


text:
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From the album The No. 1 R&B Classics Album
Released: | 60 Tracks
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Summertime By Another Level Ft Tq Rare 80s Vintage Piano Sheet Music
Tq 50mm X 50mm Promo Photo Slide Negative #c/4471
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A tasty pizza but overpriced, I guess that is Switzerland. Across the road from where I was staying at Mercure Stoller so it was very convenient for an easy option dinner.
3 Days in Zurich
One Day in Zurich
Shopping Guide for Zurich
Tasty pizza but EXTREMELY overpriced!! 19 CHF for a common pizza, that in any other place you pay 9€. They even charge you if you want to share the pizza
We have visited over 3 times in the past 4 months and at best had a drink before leaving because the place was filled with smokers smoke. A thing of the past you may think but they have a smokers room attached to the restaurant and appears to leak, or as the waiter told us, "it's the smell of pizza...
I've eaten just about everything here and it is top quality and an amazing price for Zurich. The fact the Bus and Tram drivers eat here speaks volumes.
They have a large daily menu that changes everyday except Fri-Sun when it is just Fridays menu all through. They have varied dishes all freshly prepared. I recommend the roast beef, veal...
I got a quick bite here and saved some $$ food was pretty good too.
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Akwesasne Area Management Board
Our Office Hours are Monday through Friday
8:30 a.m. to 4:30 p.m.
Phone: 613-575-2626 Fax: 613-575-2863
Toll Free Phone: 855-575-2626 Website: www.aamb.ca
2015 SEED Employer Program Guide
2015 Summer Student Employment Package
Old Age Security (OAS) Pension
Canada Pension
Social Insurance Numbers
Employment Insurance
A Service Canada Representative is available on the second Tuesday of each month at the Cornwall Island Administration Building #3 (CIA#3) from 9:30 a.m. to 3:30 p.m. (closed for lunch from Noon- 1p.m.) Please go directly to the CIA#3 for services. The next visits will be on:
- April 14
- May 12
- June 9
- July 14
- August 11
- September 8
- October 13
- November 10
- December 8
They are also available on the fourth Tuesday of each month in the village of St. Regis at the Akwesasne Area Management Board office (AAMB) from 9:30 a.m. to 3:30 p.m. ( closed for lunch from Noon – 1 p.m.) The next visits will be on:
- April 28
- May 26
- June 23
- July 28
- August 25
- September 22
- October 27
- November 24
- December 22
Please remember that if you are coming in for a SIN Card, you will need your original Birth Certificate, Status Card (also $10 for replacement cards) on hand. For any other inquiries please bring all correspondence you received from Service Canada with you.
For more information, or to make a future appointment with Service Canada, please contact the Akwesasne Area Management Board office at (613) 575-2626.
ONLINE Courses now available!
ALCN Online Courses provide access to an online learning portal that includes a library of educational courses created by InfoSource Learning, a top North American educational online course provider. Each course is a tutorial for subscribers who can learn at their own pace 24 hours a day, seven days a week at home, work, or on the go using any computer with an internet connection.
Due to a change in the mandate from the Board's Previous:
Aboriginal Human Resource Development Agreement (AHRDA) agreement
which was focused on training and development of clients
To the new:
Aboriginal Skills and Employment Training Strategy (ASETS) funding agreement
which is focused on job readiness and employment
The Akwesasne Area Management Board has made the following Policy Changes:
- . Ontario Secondary School Diploma (OSSD) / Challenges and Opportunities (CHOPS) /
Adult Literacy Employment Program (ALEP)
- Effective June 30, 2013, the AAMB will only provide tuition and training allowance funding to OSSD clients for their last 8 credits and will DISCONTINUE funding for CHOPS and ALEP
2. Training Outside of Canada
- Effective April 1, 2013, the AAMB will no longer fund training outside of Canada
For further information or questions, please contact the AAMB@ (613) 575-2626
Do you know the difference between QPIP and Service Canada Employment Insurance Benefits? Click here to find out=>QPIP Announcement
Upcoming Workshops for 2015
Inter-Agency Workshop Registration Form 2015
Monthly Calendar of Events Page
Akwesasne Employment Resource Center Job Listings
The contents of the logo has three distinct subjects:
1. The Thunderbird
2. The Eagle
3. Feathers of the Eagle encircling both birds
The Thunderbird and Eagle are almost intertwined with one another. Existing together to give guidance and protection to both worlds. (Spirtual & Reality). The Feathers that surround the images represent a universal bond between all Indigenous people and the Creator.
AAMB logo Drawn by Roger Perkins
AAMB Website Design originally created by: Colleen Nolan in 2007
Format was changed by: Monica Peters, Seaway Websites Administrator
AAMB Website went live March 2010
It is updated and maintained by Terri Bova, N.S.T.


text: Staff at this hotel is first rate, very friendly and helpful. I felt very well taken care of. I was greeted each day as I arrived and when I left. Questions were answered promptly and efficiently. Breakfast was hot and ready each day with ample selections. Pool is a little small and was not available in inclement weather or the... More


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This Hamster Lives Large in a Tiny Mansion
This hamster named Chicken is living in the lap of luxury inside a lush doll house mansion.
Hilarious Cat Doesn't Want to Go for a Walk
Egon will not budge with a leash no matter how hard his owner tries.
Opossum's Adventure Halts Minor League Baseball Game
The marsupial ran onto the field as stadium staff chased behind.
Pit Bull Showers Baby in Kisses
Video is predictably adorable.
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DAVID DRISKELL Creative Spirit: Five Decadesby Bridget Goodbody
A curator, painter, teacher, historian, and builder of private art collections (including those of Bill Cosby and Oprah Winfrey), David Driskell has spent a lifetime using the language of art to reflect African-American history. This retrospective of his own work, a celebration of the artist's 80th birthday, is ripe with this legacy. His early pieces signal Driskell's place among a group of prominent African-Americans who, like him, came of age during Martin Luther King Jr.'s 1960s. In "Jazz Singer (Lady of Leisure, Fox)" (1974), for example, the singer's face brings to mind that of Sugar as played by Margaret Avery in Steven Spielberg's 1985 film adaptation of Alice Walker's The Color Purple (1982). There are instances when Driskell's painting rewrites art history. His collage technique, used in "Jazz Singer," borrows from that of Romare Bearden, whom Driskell promoted to ensure Bearden's inclusion in the canon of American art history. Driskell also deserves a place in the pantheon of the Pattern and Decoration movement. See, for example, works like "Our Ancestors, Festival" (1973), with its painterly references to Kuba cloth and Islamic calligraphy wrapped up in Driskell's distinct, just-south-of-primary palette. All to say that Driskell's life's work is made manifest in his art. Thank you, Mr. Driskell, for using art to make our world more inclusive and more vibrant.
ContributorBridget Goodbody
SARAH LEWIS with Phong Bui
FEB 2015 | Art
In reading Sarah Lewiss The Rise I was at once reminded of how deeply appreciative I am of having read Freud, Jung, Marie-Louise von Franz, Karen Horney, Erik H. Erikson, Sir Anthony Storr, even Paul Tillich, among others. Just before the publication of the paperback edition of The Rise from Simon & Schuster, the writer/critic/curator paid a visit to the Rail HQ in Greenpoint, Brooklyn, to talk about her book with Rail publisher Phong Bui.
Magnificent Obsession: Joann Gedney, The Early Paintings, 1948 1963by Alessandro Cassin
MAY 2014 | ArtSeen
Magnificent Obsession: Joann Gedney, The Early Paintings, 1948 1963, curated by Gregory de la Haba at Rox Gallery, is an art historians dream come true: the discovery of a pristine body of work documenting the life and oeuvre of an unknown Abstract Expressionist. In addition to the paintings and drawings selected for the show, there are scores of others in the painters Village townhouse, as well as boxes of diaries, notes, and letters that complete a rich background and historical context.
by Richard Kalina
MEL BOCHNER Strong Language
JUNE 2014 | ArtSeen
Mel Bochner, one of the founders of the Conceptual Art movement of the mid-60s, and quite possibly the most inventive, clear-headed, and thought-provoking artist of that group, is showing his language-based paintings and drawings this spring and summer at the Jewish Museum.
Reading Ad Reinhardt in Polandby Edyta Frelik
AD REINHARDT | Ad Around the World
I came to Ad Reinhardts art via his writings, not his paintings. In 2000 I was working as an English teacher at various local language schools in Lublin when I was offered a position at Maria Curie-Skłodowska University.


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Welcome to Dayspring Pens!
The 'Write Place' for Personalized Cross Pens
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Jobs Watch: Brewing up success
|| Print |
|Ben Jacklet
|Wednesday, January 06, 2010
UPDATE: Figures released today by the Oregon Brewers Guild show that 2009 was the best year ever for craft brewers. For the first time, total production topped a million barrels of beer, a 15% increase over 2008.
It's always good to start the New Year on a positive note, so let's dispose of the leftover faux champagne and talk Oregon beer. The Eugene Register Guard reported yesterday that Ninkasi Brewing has become the first Oregon brewer to earn regional brewery status in a decade. Ninkasi, which added 10 jobs in 2009, is on track to crank out 30,000 barrels of beer this year, about 10 times the amount it brewed when it first launched just three years ago. By crossing the threshold from brewpub to regional brewer, this growing business joins Oregon icons such as Rogue Ales, Full Sail Brewing and Deschutes Brewery in an industry as resilient as it is quirky.
Ben Jacklet is managing editor of Oregon Business.
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|Did airlines collude to keep fares high?
|Citigroup analyst thinks Puma should sell
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Tonkon Torp helps seed sustainability at Gunderson.
Oregon-based Environments helps companies create inspired workspaces. "Simply put, we help companies future-proof their workspaces," says Chris Corrado, president. Since 1988,Environments has witnessed firsthand the changing landscape of business. Native Portlander and Environments founder Corrado says, "We help our clients navigate the complex realities of the workplace today and plan for their future in a very mindful, strategic way. We think of ourselves as their partners in the process."
One hundred years ago, the Willamette River might easily have been mistaken for a sewer. Unchecked industrial activity and decades of pollution made it unrecognizable compared to the clean river that now flows north for 187 miles from Eugene through the center of Portland.
3 Degrees Event Celebrates 5th Year Bringing Nonprofit and Business Professionals Together to Benefit Portland.
Bend energy leader brings passion for efficiency and renewable energy to the nonprofit.
Event in Forest Grove marks recognition of Global Food Safety Initiative Certification.


text: The location is extremely convenient to everything in Vienna by metro. It is just outside the ring of the city. We were happy to use the efficient metro system to get to everything and be outskirts for the night. Our room was huge for just the two of us. It was clean and the furnishings were not modern, but functional.... More


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Greenwood School, Inc.
Rating: 0 stars 0 0 review
Nonprofit Issues:
Arts & Culture, Education
14 Greenwood Ln Putney VT 05346 USA
Established in 1978, the Greenwood School is an ungraded, pre-preparatory boarding school for bright boys, ages 10-13 at the time of admittance, with dyslexia or a related language disability. Greenwood uses innovative approaches to help students bridge the gap between their potential and their present achievement. Through successful learning experiences, students master both academic and social skills. Enrollment is 40. Class sizes range from 2-10 students, with an average student-teacher ratio of 3:1. All students participate in language tutorials that remediate weaknesses while striving to help them develop their strengths. Art, drama, wood and metal working, and sports programs enable students to discover and refine their creative abilities.
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Behind The Mic: CJ's Drawer [VIDEO]
Let's take a look inside CJ's drawer here in the KTDY control room...
Let's take a look inside CJ's drawer here in the KTDY control room...
Just when you thought it was safe to answer your door...
In case you haven't heard already, Lafayette, LA is the happiest place to live in the USA!
The Cajun Jam is a weekly showcase for Acadiana's local Cajun musicians of all skill levels.
It's time to STUFF THE BUS and help local students start school with the tools they need to succeed!
Are you a member of the Carencro High Class of 1989? If so, your reunion is right around the corner!
Dr. Kevin offered insight into those often-confusing nutrition labels on food packages in this week's installment of Wellness Wednesdays.
The U.S. is a huge country, with a diverse population. It's not surprising, people from various parts of the country have stereotypes about the folks in other regions.
Here are 10 stereotypes about Louisianans that are actually right on the money.
Music, food, royalty, performing arts, and more! It's just another awesome weekend in Acadiana!
Cool air is around the corner, and that means festivals like the 2014 Louisiana Seafood Festival, held at the festival grounds in New Orleans City Park.
Ghost stories and tall tales abound in Cajun culture.
It's always good to see other parts of the country singing praises about Acadiana, and this article in the New York Daily News does just that.


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Three years after 9/11.
When the "scrawny boy from Austria" delivered his peroration against faint-hearted "economic girlie men," it was an unusually seductive, even witty, appeal to a notion of free enterprise that is
Labor Day has never been a very inspiring holiday, established as it was by late-nineteenth-century union bosses as a homegrown alternative to May Day, which was viewed as having uncomfortably le
Hidden in a Census Bureau report on poverty released in late August is a factoid with significant political and social consequences. Poverty has moved to the suburbs.
For two years, journalist Andrew Rice lived in Uganda as a fellow of the Institute of Current World Affairs.
Missouri Governor Bob Holden learned how volatile globalization issues have become when his Democratic primary challenger, Claire McCaskill, started banging away on him for offshoring the state's
The effort to rebuild Iraq looks less like an aid mission than a criminal racket.
Regular people, just plain working folks
Always stood out in my eyes.
I scorn all those people who drink wine instead
Of eating Frito pies.
As bad as Bush's economic record is, it would appear far worse if not for the housing bubble.
About half of the largest ballot-counting corporation, Election Systems & Software (ES&S), is owned by the Omaha World-Herald Company, which publishes the city's conservative daily newspa


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So new to this!!!
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Join Date: Apr 2009
Location: Canada
Posts: 7,661
It might just be that this scenario doesn't work for you. If you aren't getting enough time and require more bonding then maybe you need another partner, or need to find someone that isn't as busy and keep him in your life as a really good friend or intimate friend (FWB).
is a link to a thread about how many is enough when it comes to partners. On average it seems that in order to even get a word in edge wise three to four is plenty... or less.
Most seem to do fine with two partners before coming apart at the seams due to lack of alone time to take care of oneself. This man would rarely have time for himself. I would hate to think if there were any major issues that come up. The whole balancing plate would tumble no? All those partners with needs and expressing them... ahhhh... overwhelming.
This is worrisome to me. I have three partners, four until recently, and I struggled with looking after my primary, myself. I still see that as my number one struggle. After several years of having many partners its easy to reach burn out. I would be very careful of that. Especially as he is asking you to share date time in order to kill two birds with one stone. hmmmm, not good... smells of burn out to me (having been there)
I would wonder why he needs so many partners and what "partner" means to him. Usually a persons needs are met with far less. Did he actually seek you out? Was he looking to date more? Is it a fetish of his to collect partners?
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No foul play in hanging death
An investigation has ruled out foul play in the death of a local man whose body was found at Mhoon Landing on April 20.
Just after 10 a.m. Monday, the Tunica County Sheriff's Department responded to a call from an employee at the MBI Construction site, who reported a body hanging. Sheriff K.C. Hamp contacted the Mississippi Bureau of Investigation and the Crime Scene Unit to assist at the scene.
The body, identified as Gregory Allen Smith, 44, was transported to the Medical Examiner's office in Jackson, Miss. Smith was a resident of Plum Road at the Tunica Cut-off.
Student diagnosed with meningitis
A seven year old student at Tunica Elementary School is being treated at Lebonheur hospital in Memphis for bacterial meningitis.
School officials have taken steps to disinfect "all topical surfaces and common areas," according to a social media post by Tunica County Superintendent Steve Chandler late Tuesday.
"We are working in close collaboration with the MS Department of Health, who will provide treatment for any student or staff member who has been in close contact or shows symptoms.
"These students and staff members will be treated tomorrow, April 22, at Tunica Elementary School," Chandler said.
AG weighs in on gaming tax issue
Mississippi Attorney General Jim Hood has intervened in Tunica County, Mississippi v. Town of Tunica, et al, now pending in Circuit Court here.
A March 31 letter from the Attorney General's office asked Circuit Clerk Sharon Reynolds to file the Motion to Intervene, which reads:
Comes now Jim Hood, Attorney General of the State o Mississippi, ex rel. the State of Mississippi, pursuant to Miss. R. Civ P. 24(d) and Miss. Code Ann. Section 7-5-1, and moved this Court for permission to intervene in the above styled and numbered cause, and in support thereof would show the Court the following, to wit:
Candidate Profile: 'Customer service' anchors Dace campaign
Former math teacher, coach, bus driver, and administrator in Tunica County School Districts, Derrick "Coach" Dace is now seeking to use his experience and skills to lead the county's sole public school system in a positive direction filled with positive experiences as Superintendent.
His priorities all hinge upon offering excellent customer service to improve partnerships, academic achievement, and accountability and transparency throughout the district, he shared.
"I'm a big believer in customer service. Our customers are going to be our students, our parents, and our community," he said.
Dace shared that he plans to focus on the "PIE" to improve partnerships. He explained that "PIE" stands for "Parents, Industry, and Educators." Getting each person in these groups to work together is his goal, he added.
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Lili Jia
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2010 – today
- 2014
- [j1]Erfu Wang, Qiang Liu, Lili Jia, Qun Ding:
A steady-state point capture-based blind harmonic signal extraction algorithm under the chaotic background. IJSNet 15(1): 52-61 (2014)
- [c4]Lili Jia, Jiguo Yu, Dongxiao Yu, Qingbo Zhang:
2-m-Domatic Partition in Homogeneous Wireless Sensor Networks. WASA 2014: 378-388
- 2010
- [c3]Zhiyuan Li, Lili Jia, Xuechen Dong:
Research of Supplier Selection Based on Supply Chain Management. DBTA 2010: 1-4
- [c2]Xin Yang, Junlin Zhu, Qian Wang, Wenfang Lin, Min Chen, Lili Jia, Muwen Zhu:
The application of the spatial overlay method in regional planning of agricultural function of Hubei Province. Geoinformatics 2010: 1-4
2000 – 2009
- 2009
- [c1]Lili Jia, Huiping Zhou, Haifang Zhou, Chengzheng Sun, Zhiying Wang:
Integration of the dynamic, mutual evaluating and role-based iterated revision belief into collaborative conceptual modeling. CTS 2009: 417-422
Coauthor Index
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last updated on 2015-02-03 17:37 CET by the dblp team