Acclaim HILIC

Acclaim HILIC-10 is similar to Acclaim Mixed-Mode WAX-1. This relatively new column is also based on high-purity spherical silica covalently modified with a proprietary hydrophilic layer. It is suited for use in a broad range of applications including the separation of hydrophilic drugs and their metabolites. Acclaim Mixed-Mode HILIC-1 is a stationary phase that combines both RP and HILIC properties. Unlike traditional RP or HILIC columns, this packing material features a long alkyl chain with a diol functionality at the terminal end. 

Figure 7.21 shows a typical separation of hydrophilic pharmaceutical drugs and some degradation compounds such as salicylic acid on Acclaim HILIC-10 using UV detection at 230 nm. With a mobile phase consisting of acetonitrile and ammonium acetate at pH 5, all five components can be separated to baseline within a reasonable time. Penicillin G is an antibiotic that belongs to the -lactam fomily it is often formulated in the potassium salt form. Because both the API... 

Figure 7.21 Separation of hydrophilic pharma- acetate, pH 5 flow rate 1 mUmin detection ceuticals on Acclaim HILIC-10 under isocratic UV (230nm) injection volume 2 pU sample conditions. Column dimensions 150 mm x4.6 0.1 mg/mt of acetaminophen (1) and salicylic...

Figure 10.256 HILIC separation of meiamine and cyanuric acid with charged aerosoi detection. Separator column Acclaim HILIC-10,...

To Study the retention behavior of both Acclaim Trinity columns in the HILIC mode, three highly hydrophilic compounds with different charge states can be used as test probes weso-erithritol (neutral), Tris (cationic), and glyceric acid (anionic). 

The four Acclaim Mixed-Mode columns mentioned in Section 6.7.2 - Acclaim Mixed-Mode WAX-1, Acclaim Mixed-Mode WCX-1, Acclaim Trinity PI, and Acclaim Trinity P2 - can also be used under HILIC conditions. Acclaim Mixed-Mode WAX-1 and Acclaim Mixed-Mode WCX-1 are based on an alkyl chain with terminal ion-exchange functional groups. Because of this column chemistry, both columns offer multiple retention mechanisms, including ion exchange, reversed phase, and HILIC. As an example. Figure 7.7 demonstrates that in highly organic mobile phase. Acclaim Mixed-Mode WCX-1 can serve as a HILIC column. Hydrophilic analytes such as cytosine elute later than naphthalene as a hydrophobic neutral analyte. 

Figure 7.7 RP mode versus HILIC mode on Acclaim Mixed-Mode WCX-1. Column dimensions 1S0mmx4.6mm i.d. column temperature 30°C eluent MeCN/NH40Ac, pH 5 (5 mmol/L total), 95 5 v/v for HILIC mode...

Table 7.6 summarizes the structural and technical properties of all currently available Acclaim Mixed-Mode columns that can be used under HILIC conditions. 

Figure 7.8 Comparison of the chromatographic behavior between Acclaim Mixed-Mode HILIC-1 and LiChrosorb Diol. Separator columns Acclaim Mixed-Mode HILIC-1,5 pm ( ) and LiChrosorb Diol, 5 pm ( ) column...

Figure 7.9 Comparison of the chromatographic behavior of pharmaceuticai counterions between Acclaim Trinity PI (a) and ZIC-HILIC (b). Separator columns lOOmmxSmm i.d. Acclaim Trinity PI, 3 pm, and 150mmx4.6 mm i.d. ZIC-HILIC, 5 pm coiumn temperature 30 °C eiuent (A) 60 40 (v/v) MeCN/0.02 moi/L NH4OAC, pH 5 and eiuent (B) 80 20 (v/v) MeCN/0.015 moi/L NH4OAC, pH 5 flow rates ...

Figure 7.11 Comparison of the chromatographic behavior of a hydrophobic acidic API and its counterion between Acclaim Trinity PI (a) and ZIC-HILIC (b). Separator columns ...

Figure 7.16 Separation of selected amino acids under HILIC conditions on Acclaim Mixed-Mode WAX-1. Column dimensions 150mmx4.16mm i.d., 5pm column temperature 30 °C eluent 75 25 (vAr) MeCN/25mmol/L...

Figure 7.22 Separation of penicillin G on Acclaim Trinity PI under RP and HILIC conditions. Column dimensions SOmmxSmm i.d., 3 urn column temperature SOT eluent ...

Figure 7.24 Separation of seiected Good s buffers on Acclaim Trinity PI under HILIC conditions. Column dimensions lOOmmxSmm i.d., 3 pm column temperature 30 °C eluent 85 15 (v/v) MeCN/12 mmol/L ammonium...

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