Chromatography compositions and methods for producing same

Abstract

The chromatography composition includes a solid phase substrate and an ionically modified hydrophilic ligand attached to the solid phase substrate, the ionically modified hydrophilic ligand including a hydrophilic ligand moiety covalently attached to the solid phase substrate, the hydrophilic ligand including a polar group and a plurality of hydroxyl groups, the ionically modified hydrophilic ligand also including an ionic group attached directly or indirectly to the hydrophilic ligand moiety. A method for producing the chromatography composition is also provided.

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application claims priority to and all benefits of U.S. Provisional Patent Application No. 63 / 341,617, filed May 13, 2022, the entire contents of which are incorporated herein by reference.

[0002] (Technical field) The present disclosure relates generally to chromatographic compositions for use in chromatographic separations. [Background technology]

[0003] High performance liquid chromatography (HPLC) is a process for separating components in a liquid mixture. There are many different forms of HPLC, including ion exchange, reversed phase, and hydrophilic interaction liquid chromatography (HILIC), and mixed-mode hydrophilic interaction liquid chromatography with ion exchange properties. Each of these variants contains a mobile phase and a stationary phase that work together to achieve separation. Despite the general ability of liquid chromatography to retain and separate polar analytes, traditional stationary phases are not optimized to separate certain mixtures of low molecular weight, highly polar substances, such as perfluoroalkyl substances (PFAS). Thus, opportunities remain for improving chromatographic compositions. Summary of the Invention [Means for solving the problem]

[0004] In one aspect of the disclosure, a chromatography composition is provided. The chromatography composition includes a solid phase substrate and an ionically modified hydrophilic ligand bound to the solid phase substrate. The ionically modified hydrophilic ligand includes a hydrophilic ligand moiety covalently bound to the solid phase substrate with the hydrophilic ligand including a polar group and a plurality of hydroxyl groups. The ionically modified hydrophilic ligand also includes an ionic group directly or indirectly bound to the hydrophilic ligand moiety.

[0005] In another aspect of the disclosure, a method for preparing a chromatographic composition is provided. The method includes providing a solid phase substrate and providing a hydrophilic ligand comprising a polar group and a plurality of hydroxyl groups, with at least one hydroxyl group present at a terminal end of the hydrophilic ligand. The method also includes reacting the solid phase substrate with the hydrophilic ligand to covalently bond the hydrophilic ligand to the solid phase substrate to form a hydrophilically modified substrate. The method further includes providing an activated compound comprising a leaving group, and reacting the activated compound with the terminal hydroxyl group of the hydrophilic modified substrate to form an activated hydrophilic modified substrate. The method further includes providing an ionic modifier comprising a nucleophile and an ionic group, and reacting the activated hydrophilic modified substrate with the ionic modifier to release the leaving group of the activated compound and form an ionic modified hydrophilic ligand and a chromatographic composition.

[0006] In another aspect of the disclosure, a second method of producing a chromatography composition is provided. The method includes providing a solid phase substrate and providing a hydrophilic ligand comprising a polar group and a plurality of hydroxyl groups, such that at least one hydroxyl group is present at a terminal end of the hydrophilic ligand. The method also includes reacting the solid phase substrate with the hydrophilic ligand to covalently bond the hydrophilic ligand to the solid phase substrate to form a hydrophilically modified substrate. The method further includes providing an activating compound comprising a leaving group, and providing an ionic modifier comprising a nucleophile and an ionic group. The method further includes reacting the activating compound with the ionic modifier to form an activated ionic compound. The method further includes reacting the terminal hydroxyl group of the hydrophilically modified substrate with the activated ionic compound to form an ionically modified hydrophilic ligand.

[0007] The chromatographic composition is useful for chemical separation. For example, the chromatographic composition is useful as a stationary phase in HPLC separation, such as ion exchange chromatography, HILIC, and mixed mode HILIC. Without being bound by any particular theory, it is believed that ion-modified hydrophilic ligands provide superior separation ability in HPLC separation of selected analytes, such as PFAS, when compared with chromatographic compositions that include hydrophilic ligands that are not ion-modified. [Brief description of the drawings]

[0008] The advantages of the present invention will be better understood and readily appreciated by reference to the following detailed description taken in conjunction with the accompanying drawings, in which:

[0009] [Figure 1] Chromatogram showing separation of low molecular weight PFAS compounds by a conventional chromatographic composition containing hydrophilic ligands bound to the surface of silica. [Diagram 2] Chromatogram showing separation of low molecular weight PFAS compounds by a chromatographic composition containing ionically modified hydrophilic ligands bound to a silica surface at a density of 0.6 μmol / m2. [Diagram 3] Chromatogram showing separation of low molecular weight PFAS compounds by a chromatographic composition containing ionically modified hydrophilic ligands bound to a silica surface at a density of 1.2 μmol / m2. [Figure 4] Chromatogram showing separation of low molecular weight PFAS compounds by a chromatographic composition containing ionically modified hydrophilic ligands bound to a silica surface at a density of 1.7 μmol / m2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] The present disclosure provides a chromatographic composition.The chromatographic composition is useful for chemical separation, particularly for HPLC separation, comprising a stationary phase and a mobile phase.In certain embodiments, the chromatographic composition is particularly useful as the stationary phase for ion exchange chromatography, HILIC, and mixed-mode HILIC.

[0011] The chromatographic composition comprises a solid phase substrate and an ionically modified hydrophilic ligand bound to the solid phase substrate. In certain aspects, bound to the solid phase substrate means covalently bound to the solid phase substrate. The ionically modified hydrophilic ligand comprises a hydrophilic ligand moiety covalently bound to the solid phase substrate and an ionic group directly or indirectly bound to the hydrophilic ligand moiety. The hydrophilic ligand moiety comprises a polar group and a plurality of hydroxyl groups.

[0012] The polar group of the hydrophilic ligand moiety can be selected from carbonate, carbamate, amide, amine, ureido, ether, thioether, sulfinyl, sulfoxide, sulfonyl, thiourea, thiocarbonate, or thiocarbamate, and can include heterocyclic compounds containing polar functional groups. For example, the polar group can be an aromatic ring containing an amine. In one embodiment, the polar group X is selected from amide or carbamate. The multiple hydroxyl groups present on the hydrophilic ligand moiety can be two or more hydroxyl groups. Alternatively, the hydrophilic ligand moiety can include 2 to 8, 2 to 7, or 3 to 5 hydroxyl groups.

[0013] First, the solid phase substrate is typically, but not necessarily, silica. The silica used in the chromatography composition is not limited to a particular grade. Both non-porous spherical silica and porous silica (including superficially porous silica) can be used. Silica particles typically have an average particle size of 0.5-100 μm, 1-50 μm, 1.5-10 μm, or 1.7-5 μm. Porous silica can have an average pore size of about 80 Å or more, about 250 Å or more, about 300 Å or more, about 450 Å or more, 200 to 1,000 Å, 250 to 900 Å, or 300 to 850 Å. Alternatively, it is expected that the average pore size can be about 1 to about 50 Å, about 5 to about 40 Å, or about 10 to about 30 Å, although pore sizes less than 70 Å are generally avoided. The surfaces of silica particles generally contain silica hydroxyl groups (so-called silanols), which are useful for covalent attachment of various reagents to the silica surface. In most cases, specific organosilane reagents are used for these silica surface modifications to form a covalently bonded bonded phase. Suitable grades of silica are available from Advanced Materials Technologies, headquartered in Wilmington, Delaware, under the trade name Halo Silica, but many silica materials are widely available as commercial materials for a variety of useful applications. Alternative substrates include hybrid inorganic / organic materials. In the context of this disclosure, the term "hybrid inorganic / organic materials" includes inorganic-based structures in which organic functional groups are integral both in the interior (i.e., on the hybrid material surface as well as in the inorganic structure). The inorganic portion of the hybrid material can be, for example, alumina, silica, titanium, cerium, or zirconium, or oxides thereof, or ceramic materials. Yet other substrates include fully organic substrates, which contain hydroxyl groups on the surface of the organic substrate. For the purposes of this disclosure, the solid phase substrate is not formed from carbohydrates. However, carbohydrates may be included when covalently attached to inorganic or hybrid inorganic / organic materials.

[0014] Although not required, the ionically modified hydrophilic ligand may be of formula I: [ka] (where: X is a polar group, Z is a polar linking group; Y is an ionic group; n is 1-6, n' is 0-2, m is 2-8, p is 0 or 1, s is 1, R 1 , R 2 , R 3 are independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group, and R 4 and R 5 is independently H or OH, and at least two m units contain at least one hydroxyl group. It can be derived from

[0015] The ionically modified hydrophilic ligand is attached to the solid substrate, so that it can be coupled to the surface hydroxyl groups present on the solid substrate and the three [(R 1 It is understood that the reaction occurs between one of the (O) units. Thus, the ionically modified hydrophilic ligand is derived from and represented by Formula I prior to reaction with the solid phase substrate.

[0016] When the ionically modified hydrophilic ligand is represented by formula I, the hydrophilic ligand moiety has formula Ia: [ka] (X is a polar group; n is 1-6, and n' is 0-2, R 1 , R 2 , R 3 is independently H or a straight or branched, substituted or unsubstituted, C1 to C18 alkyl group; and R 4 and R 5is independently H or OH, and at least two m units contain at least one hydroxyl group. is derived from.

[0017] Typically, but not necessarily, p is 1, such that a polar linking group is present on the ionically modified hydrophilic ligand.

[0018] The polar group X is independently selected from carbonate, carbamate, amide, amine, ureido, ether, thioether, sulfinyl, sulfoxide, sulfonyl, thiourea, thiocarbonate, or thiocarbamate, and includes heterocyclic compounds containing polar functional groups. For example, the polar group can be an aromatic ring containing an amine. In one embodiment, the polar group X is selected from amide or carbamate. In another embodiment, the polar group X is an amide. When the polar group X is an amide, the ionically modified hydrophilic ligand is represented by formula Ib: [ka] It can be derived from

[0019] In certain embodiments of Formula I and Formula Ib, n is 2-4, m is 3-6, p is 1, and R 1 , R 2 , R 3 are independently H or a straight or branched, substituted or unsubstituted C1-C6 alkyl group. When X is amide, n' is typically 0, although not required. In other embodiments of Formula I, when X is ureido, n' is 1 or 2. In one embodiment of Formula Ib, n is 3, X is amide, m is 5, and four of the m units contain only one hydroxyl group. In one embodiment, the ionically modified hydrophilic ligand is represented by Formula Ic: [ka] is derived from.

[0020] In certain embodiments, p is 1 and a polar linking group Z is included in the ionically modified hydrophilic ligand. Although not required, the polar linking group Z is typically a carbamate group when p is 1. In certain embodiments of formula Ic where the polar group X is an amide, a polar linking group Z is also present, m is 5, four of the m units contain only one hydroxyl group, and the ionically modified hydrophilic ligand has formula II: [ka] is derived from.

[0021] When the polar linking group Z is a carbamate group, formula II can be further represented by formula IIa: [ka] It is expressed as:

[0022] Referring now to the ionic group Y, typically the ionic group Y is derived from an ionic modifier that includes a nucleophile and an ionic group (described further below). The ionic group Y can be represented by formula III: [ka] (k is 1-8, R 6 and R 7 is independently H or a straight or branched chain, substituted or unsubstituted, C1 to C18 alkyl group, which may also include halocarbon or alcohol substituents; and I is a charge-carrying functional group capable of (1) carrying a positive ionic charge under neutral or acidic aqueous or aqueous organic solvent conditions, or (2) carrying a negative ionic charge under suitable neutral or basic aqueous or aqueous organic solvent conditions. It can be expressed as:

[0023] Those of skill in the art will understand that the phrase "capable of" in the context of carrying an ionic charge means that the ionic group is either positive or negative when exposed to the aqueous or aqueous organic solvent conditions described above. Those of skill in the art will also understand that the ionic group need not be cationic or anionic if it is not exposed to liquid conditions.

[0024] In some embodiments, the ionic group I, which can bear a positive charge, is a primary, secondary, tertiary, quaternary amine or aromatic amine. When the ionic group I is a primary, secondary, tertiary, quaternary or aromatic amine, the nitrogen atom of the amine can be bonded to hydrogen, alkyl, alcohol-substituted alkyl, aromatic groups, and combinations thereof. In other words, the ionic group I can be bonded to hydrogen, alkyl, alcohol-substituted alkyl, and aromatic groups at two or more positions and is included as part of a heterocyclic compound. In some embodiments, the ionic group is a tertiary amine and is bonded to two ethyl groups. In other embodiments, the ionic group I is part of a heterocyclic compound, and I is [ka] It could be.

[0025] In another embodiment, the ionizable group capable of carrying a negative charge is an immobilized carboxylic or sulfonic acid.

[0026] With further reference to Formula III, typically k is 2-6 and R 6 and R 7 is hydrogen.

[0027] In certain embodiments of the ionically modified hydrophilic ligand, the polar group X is an amide, Z is present and is a carbamate group, m is 3-7, n is 2 to 4, n' is 0 or 1, at least four m units contain only one hydroxyl group, and the ionic group Y comprises a tertiary amine. In another embodiment, the ionically modified hydrophilic ligand has formula IV: [ka] is derived from.

[0028] Returning to the chromatographic composition generally, in addition to having an ionically modified hydrophilic ligand bound to the solid phase substrate, the chromatographic composition may also have a hydrophilic ligand bound and / or covalently attached to the solid phase substrate, In other words, when a hydrophilic ligand is present along with an ionically modified hydrophilic ligand, the hydrophilic ligand is not ionically modified.

[0029] The polar group of the hydrophilic ligand may be selected from carbonate, carbamate, amide, amine, ureido, ether, thioether, sulfinyl, sulfoxide, sulfonyl, thiourea, thiocarbonate, or thiocarbamate, and may include a heterocyclic compound containing a polar functional group. For example, the polar group may be an aromatic ring containing an amine. In one embodiment, the polar group is selected from an amide or a carbamate. The multiple hydroxyl groups present on the hydrophilic ligand may be two or more hydroxyl groups. Alternatively, the hydrophilic ligand may include 2 to 8, 2 to 7, or 3 to 5 hydroxyl groups.

[0030] In one embodiment, the hydrophilic ligand has formula V: [ka] (X is a polar group, n is 1-6, n' is 0-2, m is 2-8, q is 1, R 1 , R 2 , R 3 is independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group; R 4 and R 5 is independently H or OH, and at least two m units contain at least one hydroxyl group; and R8 and R 9 R is independently H or OH; 8 and R 9 At least one of is OH.) is derived from.

[0031] R in unit q 8 and / or R 9 The hydroxyl group of the ionically modified hydrophilic ligand is also referred to as a terminal hydroxyl group. A person skilled in the art will recognize that the ionically modified hydrophilic ligand portion of the ionically modified hydrophilic ligand and the hydrophilic ligand are the units q (i.e., -[C(R 8 )(R 9 )] q It will be understood that the hydrophilic ligands share a similar structure except that they do not contain a q unit and therefore do not contain a terminal hydroxyl group. Thus, the hydrophilic ligands can include each of the various structural configurations of the hydrophilic ligand moieties described above, except that the hydrophilic ligands further include a q unit.

[0032] In one embodiment, the hydrophilic ligand of formula V may further be represented by formula Va: [ka] is derived from

[0033] When the chromatographic composition includes a hydrophilic ligand in addition to the ionically modified hydrophilic ligand, the relative amount of each ligand can be optimized based on the particular analyte to be separated. For example, in certain embodiments, the ionically modified hydrophilic ligand and the hydrophilic ligand are present in a molar ratio range of 1:10 to 10:1. Alternatively, the ionically modified hydrophilic ligand and the hydrophilic ligand can be present in a molar ratio range of 2:8 to 8:2, 3:7 to 7:3, 4:6 to 6:4, or about 1:1. In certain embodiments, the solid phase substrate is a superficially porous silica covalently bonded to an ionically modified hydrophilic ligand represented by Formula I and a hydrophilic ligand represented by Formula V. Alternatively, in one embodiment, the solid phase substrate is a superficially porous silica covalently bonded to an ionically modified hydrophilic ligand represented by Formula Va and a hydrophilic ligand represented by Formula II.

[0034] The present disclosure also provides a method for preparing a chromatographic composition. The method includes providing a solid phase substrate and providing a hydrophilic ligand comprising a polar group and a plurality of hydroxyl groups. At least one hydroxyl group is present at the terminus of the hydrophilic ligand, and typically only one hydroxyl group is present at the terminus. Both the solid phase substrate and the hydrophilic ligand are described above. The method further includes reacting the solid phase substrate with the hydrophilic ligand to covalently bond the hydrophilic ligand to the solid phase substrate to form a hydrophilically modified substrate. The method further includes providing an activating compound comprising a leaving group, and preferentially reacting the activating compound with the terminal hydroxyl group of the hydrophilically modified substrate to form an activated hydrophilically modified substrate. The terminal hydroxyl group is by design a primary hydroxyl group, although in other examples, the hydrophilic ligand has a secondary hydroxyl group. This differentiation can allow for selective reaction of the primary hydroxyl group relative to the secondary hydroxyl group. In other words, the method includes a first reaction between the solid phase substrate and a second reaction between the reaction product of the first reaction (i.e., the hydrophilically modified substrate) and the activated compound. The method further includes providing an ionic modifier comprising a nucleophile and an ionic group, and reacting the ionic modifier with the activated hydrophilically modified substrate to release a leaving group of the activator and form an ionically modified hydrophilic ligand covalently attached to the solid phase substrate. In other words, the method also includes a third reaction between the reaction product of the second reaction (i.e., the reaction between the activated compound and the hydrophilically modified substrate) and the ionic modifier. The resulting reaction product of the third reaction is a chromatographic composition comprising an ionically modified hydrophilic ligand covalently attached to the solid phase substrate.

[0035] First, referring to the first reaction between the solid phase substrate and the hydrophilic ligand, the reaction is carried out by reacting a surface hydroxyl group present on the solid phase substrate with a hydrophilic ligand represented by formula V: [ka] The three [(R 1 The resulting reaction product produces a hydrophilically modified substrate and is a [C(R8 )(R 9 )].

[0036] Typically, the second reaction between the hydrophilic modified substrate and the activating compound occurs under aprotic anhydrous solvent conditions to limit hydrolytic loss of the activated complex. The activating compound may contain a carbonyl group. Specific examples of activating compounds containing a carbonyl group include, but are not limited to, phosgene (carbonyl dichloride), carbonyl diimidazole (CDI), or chloroformates such as 4-nitrophenyl chloroformate (4-NPC), or carbonates such as N,N'-disuccinimidyl carbonate (DSC), or combinations thereof. An illustrative example of the second reaction product between the hydrophilic ligand of formula Va and DSC is shown below, which forms the N-hydroxysuccinimidyl (NHS) carbonate of the 3-TPG compound. [ka]

[0037] Alternative activation compounds include, but are not limited to, compounds with a tosylate group, such as tosyl chloride (4-toluenesulfonyl chloride). Further suitable activation compounds include mesyl chloride (methanesulfonyl chloride), triphenylmethylene chloride (trityl chloride), phosphorus tribromide, or thionyl chloride. Although not typical, any reactive compound can be used in combination with the alternative activation compound.

[0038] Without being bound by any particular theory, it is believed that the activating compound described herein can selectively react with the terminal hydroxyl group of the hydrophilic ligand under suitable conditions. Selective reaction at the terminal hydroxyl group is also believed to be an important aspect of the present disclosure as homogeneity and generally avoidance of multiple reaction products (such as crosslinked intermediates or cyclic carbonates), and is also favorable for achieving consistent chromatographic separation. When the hydrophilic ligand is reacted with the activating compound, the hydrophilic ligand portion of formula Ia is fixed.

[0039] The ionic modifier has the formula VI: [ka] It can be expressed as W is a nucleophile; k is 1-8, R 6 and R 7 is independently H or a straight or branched chain, substituted or unsubstituted C1 to C18 alkyl group, which may contain halocarbon or alcohol substituents; I is a charge-carrying functional group capable of i. carrying a positive ionic charge under neutral or acidic aqueous or aqueous organic solvent conditions, or ii. carrying a negative ionic charge under suitable neutral or basic aqueous or aqueous organic solvent conditions.

[0040] In some embodiments, the ionic modifier is selected from N,N-(diethyl)-diaminoethane, 4-(aminoethyl)pyridine, 2-aminoethanesulfonic acid, 2-aminoethanesulfinic acid, and 3-aminopropanesulfonic acid. In another embodiment, the ionic group of the ionic modifier is a tertiary amine. In another embodiment, the ionic modifier is N,N-(diethyl)-diaminoethane.

[0041] An example is provided to aid in understanding the third reaction product obtained from reacting the above reaction product with N,N-(diethyl)-diaminoethane. [ka]

[0042] As described above, reaction of the second reaction product with N,N-(diethyl)-diaminoethane displaces the leaving group of the activated compound and forms a carbamate (urethane) bond. The carbamate (urethane) bond represents the polar linking group Z in Formula I. And the ionizable group of the ionic modifier is the ionizable group Y in Formula I.

[0043] The present disclosure provides another method (i.e., the second method) of producing a chromatography composition. Similar to the first method, the second method includes providing a solid phase substrate and providing a hydrophilic ligand that includes a polar group and a plurality of hydroxyl groups. At least one hydroxyl group is present at the terminus of the hydrophilic ligand, and typically only one hydroxyl group is present at its terminus. Both the solid phase substrate and the hydrophilic ligand are described above. The method further includes reacting the solid phase substrate with the hydrophilic ligand to covalently bond the hydrophilic ligand to the solid phase substrate to form a hydrophilically modified substrate. Unlike the first method, the second method includes providing an activating compound that includes a leaving group, and further providing an ionic modifier that includes a nucleophile and an ionic group, and reacting the ionic modifier with the activating compound to form an activated ion-producing compound. In other words, unlike the first method, the second method reacts the activating compound with the ionic modifier before either of these components are bound to the solid phase substrate or the hydrophilically modified substrate. Instead, the ion modifier and the activating compound are reacted "out-of-particle" and the resulting reaction product (activated ion-producing compound) is then reacted with the hydrophilically modified substrate via the terminal hydroxyl of the hydrophilic ligand moiety to form the chromatographic composition. As mentioned above, the terminal hydroxyl group is, by design, a primary hydroxyl group. As mentioned above, there are numerous options for activating the ion-producing compound to attach it to the hydrophilically modified substrate. For example, suitable activating compounds include, but are not limited to, compounds having a tosylate group, such as tosyl chloride (4-toluenesulfonyl chloride). Further suitable activating compounds include mesyl chloride (methanesulfonyl chloride), triphenylmethylene chloride (trityl chloride), phosphorus tribromide, or thionyl chloride. Although not common, any reacting compound can be used in combination with an alternative activating compound. These compounds may generally be selected to attach the ion-producing compound to the terminal hydroxyl group, which may be considered a weakly nucleophilic reaction site. A reaction scheme showing a typical reaction product between an activating compound and an ion modifier to produce an ion-producing compound is shown below. [ka] The following reaction product between an ion-generating compound and a hydrophilic ligand attached to a solid phase substrate is shown below: [ka]

[0044] As mentioned above, the chromatographic composition is useful for HPLC separation, including HILIC, mixed mode HILIC, and ion exchange chromatography. Further applications include, but are not limited to, thin layer plates, filtration membranes, microfluidic separation devices, sample clean-up devices, solid supports, solid phase extraction devices, microchip separation devices, or microtiter plates. The chromatographic composition can be included in a kit, and the kit can optionally include instructions for use of the chromatographic composition.

[0045] The method of preparing the chromatographic composition may also include binding both ionically modified hydrophilic ligands and hydrophilic ligands (i.e., non-ionically modified) to the solid phase substrate by controlling the stoichiometry of the second reaction. Specifically, after the hydrophilic ligands are covalently bound to the surface of the solid phase substrate, the hydrophilic ligands can be maintained in their current state by including a mole number of activating compound less than the mole number of hydrophilic ligands bound to the substrate. In particular, the ionizing agent may only react with the activated hydrophilic ligands and not with the hydrophilic ligands (i.e., non-activated hydrophilic ligands), so that the remaining hydrophilic ligands are maintained in an unmodified state. Alternatively, the reaction of activation and modification of the hydrophilic ligands can occur in free solution, resulting in a mixture that can then be covalently bound to the solid phase carrier.

[0046] (Example) Those skilled in the art will recognize that equivalents to the following instruments and suppliers exist, and the instruments listed below should not be construed as limiting.

[0047] Elemental analyses (%C, %H, %N) were determined by combustion analysis (Robertson Microlit Laboratories, Ridgewood, NJ). These values ​​correspond to known compound compositions and specific surface areas (m 2 The specific surface area (SSA), specific pore volume (SPY), and average pore diameter (APD) of these materials were measured using multipoint N 2 The adsorption method was measured (Micromeritics ASAP 2400, Micromeritics Instruments Inc., Norcross, GA). SSA was calculated using the BET method, and SPY was calculated as P / P 0 APD was calculated from the desorption portion of the isotherm using the BJH method, with a single point value determined to be >0.98. Particle size was measured using a Beckman Coulter Multisizer 3 Analyzer (30 μm aperture, 70,000 counts, Miami, FL). Particle size (dp) was measured as the 50% cumulative diameter of the volumetric particle size distribution. The width of the distribution was measured as the 90% cumulative volume diameter divided by the 10% cumulative volume diameter (expressed as a 90 / 10 ratio). In general, surface coverage is expressed normalized to the elemental composition and SSA of the sample and is expressed as the molar surface coverage of the silica surface with ligands (μmol / m 2 ) is obtained.

[0048] Commercially available 2.7 μm diameter fully hydroxylated superficially porous silica particles (Halosilica 25 g, Advanced Materials Technologies, Wilmington, DE, SSA = 120 m 2The silica particles were dispersed under nitrogen and refluxed in toluene (250 mL, Millipore / Sigma, St. Louis, NJ) with a Dean-Stark trap for 1 h to collect a small amount of adsorbed water. After briefly cooling to approximately 65 °C, a 12 mmol amount of diisopropylethylamine (DIPEA, Sigma-Aldrich, St. Louis, MO) was added with stirring, followed by 36 mmol of N-(3-triethoxysilylpropyl)gluconamide (3TPG, 30% in ethanol, Gel-Est Inc., Morrisville, PA). The resulting mixture was heated to 78 °C to remove most of the ethanol, and then refluxed overnight with approximately 5 mL of solvent collected at intervals to aid in the removal of ethanol released during the binding of ethoxysilane to the surface of the silica particles. After cooling, the resulting silica particles were collected by filtration through a sintered glass funnel, washed with 200 mL of warm toluene, THF, acetonitrile, then methanol (all solvents from Sigma-Millipore), dried on the filter, and then the silica was further dried in a vacuum oven at 110 °C for at least 1 h. The resulting 3-TPG-bonded silica was then subjected to an additional coupling reaction with 6 mmol of DIPEA and 18 mmol of 3-TPG in 250 mL of dimethylformamide (DMF, Sigma-Aldrich, St. Louis, MO) at a temperature of 85 °C overnight with occasional removal of about 5 mL of solvent via a Dean-Stark trap. After cooling, the solid was collected by filtration, washed with warm DMF, then acetonitrile, then dispersed in 50% acetonitrile / water, collected by filtration, and washed with acetonitrile and methanol. The silica was dried under vacuum at 110 °C as before. The resulting 3-TPG-bound silica particles are tightly bound to 3-TPG, and elemental analysis reveals that the 3-TPG bond is typically 3.5-3.8 μmol / m 2 Chromatographic analysis of the resulting material revealed typical retention characteristics of hydrophilic interaction liquid chromatography, an example of which is shown in Figure 1.

[0049] (Ion Modification) The 3-TPG-bonded silica particles of Example 1 are dried in a vacuum oven for 2 hours. An appropriate amount of 5-20 g of material is dispersed in a volume of 10 mL per gram using dry acetonitrile (Sigma-Aldrich, St. Louis, MO), and then 4-dimethylaminopyridine (DMAP, Sigma-Aldrich) in an amount of 0.2 mmol / g is added with stirring at room temperature, followed by disuccinimidyl carbonate (DSC, Oakland Chemicals) in an amount of 0.24 mmol / g (Rx3a), 0.48 mmol / g (Rx3b), or 0.96 mmol / g (Rx3c) with stirring and dispersion in an ultrasonic bath. The reaction to form the NHS-activated intermediate is allowed to proceed at room temperature under nitrogen for 1.5 hours, after which each of these reaction mixtures is kept separate and the silica particles are collected on a filter and washed with 25 mL / g dry acetonitrile, THF, 20% THF in 1 mM aqueous HCl, then THF, acetonitrile, and methanol. After drying under vacuum at room temperature, the activated 3-TPG silica is dispersed in acetonitrile at 10 mL / g silica solids with stirring, to which is added 5 mmol / g (2-aminoethyl)diethylamine and allowed to react overnight under nitrogen.

[0050] The resulting reaction product of the DEAE addition was collected by centrifugation (1000xg, 5 min), dispersed in acetonitrile, and the collection was washed twice using dispersion centrifugation with 10 mL / g acetonitrile. Hydrolysis of unreacted NHS modification sites was performed by dispersing the silica particles in a 0.2 M carbonate buffer (pH 9.5) / 10% acetonitrile solution and mixing for 30 min, followed by dispersion in 0.5 M Tris buffer (pH 7.8) for 30 min. The modified silica was then washed twice by dispersion in water and centrifugation, then dispersed in 10 ml / g water, collected, vacuum filtered, and washed on the filter with approximately 10 mL / g of acetonitrile and methanol before drying on the filter, followed by drying in a vacuum oven at 110 °C. In this example, the activation modification was performed under continued excess availability of (2-aminoethyl)diethylamine as an ion-modifying reagent. Elemental analysis of the resulting silica particles showed that varying the amount of activator treatment resulted in predictable and controlled addition of ionic DEAE groups to the surface, resulting in 0.6 μmol / m ionic DEAE groups upon DSC activation with 0.24 mmol / g, 0.48 mmol / g, and 0.96 mmol / g of TPG silica, respectively. 2 , 1.2 μmol / m 2 , 1.7 μmol / m 2 Chromatographic analysis of the resulting ionically modified materials revealed typical retention characteristics for hydrophilic interaction liquid chromatography, examples of which, together with various degrees of surface modification, are shown in Figures 2-4.

[0051] (Chromatographic separation of PFAS) 3-TPG and ionically modified 3-TPG silica were employed to pack stainless steel HPLC columns with 2.1 in. ID × 100 mm length. These columns were tested for separation of PFAS standards (Wellington Laboratories, Guelph, Ontario, Canada) by injecting 1 μL of the appropriately diluted standard mixture. Separation was performed on a Shimadzu Nexera LC instrument with a flow rate of 0.4 mL / min, column temperature of 40 °C, and a linear gradient elution program from 70% A / 30% B to 20% A / 80% B over the course of 10 min, where A consisted of 60% acetonitrile / 40% methanol with 0.1% formic acid, and B consisted of 10 mM ammonium formate / 0.1% formic acid in water. Detection of individual PFAS compounds was performed by online mass spectrometry coupled with HPLC separation using a Shimadzu LCMS-8040 triple quadrupole instrument. The results shown in the accompanying figures are the negative ion electrospray (ESI) total ion currents of the precursor fragments identified by the MS system, using the precursor masses appropriate for each compound.

[0052] In particular, three samples of chromatographic materials with ionically modified hydrophilic ligands showed increased retention of anionic analytes (ionized perfluorocarbons) depending on the degree of surface cation modification, together with significantly superior separation performance (Figures 2-4) when compared to conventional chromatographic materials (Figure 1).

[0053] It is understood that the appended claims are not limited to describing any particular compounds, compositions, or methods described in the detailed description, which may vary among specific embodiments within the scope of the appended claims. With respect to any Markush group relied upon herein to describe particular features or aspects of various embodiments, different, special, and / or unexpected results may be obtained from each member of the respective Markush group, independent of all other Markush members. Each member of a Markush group may be relied upon individually or in combination to adequately support a particular embodiment within the scope of the appended claims.

[0054] Moreover, the ranges and subranges relied upon in describing various embodiments of the present disclosure are understood to be included in the appended claims, both individually and collectively, and to describe and contemplate all ranges, including integers and / or decimals, even if not explicitly stated herein. Those skilled in the art will readily appreciate that the recited ranges and subranges fully describe and enable various embodiments of the present disclosure, and that such ranges and subranges may be further subdivided into relevant halves, thirds, quarters, fifths, etc. By way of example only, the range "0.1 to 0.9" may be further divided into a lower third, i.e., 0.1 to 0.3, a middle third, i.e., 0.4 to 0.6, and an upper third, i.e., 0.7 to 0.9, each of which is individually and collectively within the scope of the appended claims and may be relied upon individually and / or collectively to fully support a particular embodiment within the scope of the appended claims. Furthermore, with respect to words defining or modifying a range, such as "at least," "greater than," "less than," "less than," etc., it should be understood that such words include subranges and / or upper or lower limits. As another example, the range "at least 10" inherently includes the subranges of at least 10 to 35, at least 10 to 25, 25 to 35, etc., each of which may be relied upon individually and / or collectively to fully support certain embodiments within the scope of the appended claims. Finally, individual numbers within the disclosed ranges may be relied upon to fully support certain embodiments within the scope of the appended claims. For example, the range "1 to 9" includes various individual integers, such as 3, and individual numbers including decimal points (or fractions), such as 4.1, which may be relied upon to fully support certain embodiments within the scope of the appended claims.

[0055] The present disclosure has been described in an illustrative manner, with it being understood that the terminology used is intended as words of description rather than of limitation. Many modifications and variations of the present disclosure are possible in light of the above teachings. The present disclosure may be practiced otherwise than as specifically described. The subject matter of all combinations of independent and dependent claims, both singly and multiply dependent, is expressly contemplated herein.

[0056] (Additional Note) (Appendix 1) A solid phase substrate, and the solid phase substrate comprising an ionically modified hydrophilic ligand attached thereto, the ionically modified hydrophilic ligand comprising a hydrophilic ligand moiety covalently attached to said solid phase substrate, said ligand moiety comprising a polar group and a plurality of hydroxyl groups; and an ionic group attached directly or indirectly to said hydrophilic ligand moiety; Including, Chromatographic compositions.

[0057] (Appendix 2) The ionically modified hydrophilic ligand has formula I: [ka] (where: X is a polar group, Z is a polar linking group; Y is an ionic group; n is 1-6, n' is 0-2, m is 2-8, p is 0 or 1, s is 1, R 1 , R 2 , R 3 are independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group, and R 4 and R 5 is independently H or OH, and at least two m units contain at least one hydroxyl group. 2. The chromatography composition of claim 1, derived from

[0058] (Appendix 3) The hydrophilic ligand moiety has the formula Ia: [ka] (where: X is a polar group, n is 1-6, n' is 0-2, R 1 , R 2 , R 3 are independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group, and R 4 and R 5 is independently H or OH, and at least two m units contain at least one hydroxyl group. 3. The chromatography composition of claim 1 or 2, derived from

[0059] (Appendix 4) 4. The chromatographic composition of any one of claims 1 to 3, wherein the polar group X is independently selected from carbonate, carbamate, amide, amine, ureido, ether, thioether, sulfinyl, sulfoxide, sulfonyl, thiourea, thiocarbonate, or thiocarbamate, and comprises a heterocyclic compound containing a polar functional group.

[0060] (Appendix 5) 5. The chromatographic composition of claim 4, wherein the polar group X is selected from an amide, a carbamate, or a ureido group.

[0061] (Appendix 6) 5. The chromatography composition of claim 4, wherein the polar group X is an amide.

[0062] (Appendix 7) n is 2-4, m is 3-6, p is 1, and R 1 , R 2 , R 3 is independently H or a straight or branched, substituted or unsubstituted C1 to C6 alkyl group; 7. The chromatography composition of any one of claims 1 to 6.

[0063] (Appendix 8) 8. The chromatography composition of any one of claims 1 to 7, wherein the polar linking group Z is a carbamate group and p is 1.

[0064] (Appendix 9) The ionically modified hydrophilic ligand has formula II: [ka] 9. The chromatography composition of any one of claims 1 to 8, derived from

[0065] (Appendix 10) The ionic group Y is represented by formula III: [ka] (where: k is 1-8, R 6 and R 7 are independently H or a straight or branched chain, substituted or unsubstituted C1 to C18 alkyl group, which may also contain halocarbon or alcohol substituents; I is a charged functional group, i. bears a positive ionic charge under neutral or acidic aqueous or aqueous organic solvent conditions; or ii. bears a negative ionic charge under suitable neutral or basic aqueous or aqueous organic solvent conditions; It is possible.) 10. The chromatography composition of any one of claims 1 to 9, wherein

[0066] (Appendix 11) 11. The chromatography composition of claim 10, wherein I is a primary, secondary, tertiary, or quaternary amine.

[0067] (Appendix 12) 12. The chromatography composition of claim 11, wherein the nitrogen atom of the amine is bonded to a hydrogen, an alkyl, an alcohol-substituted alkyl, an aromatic group, and combinations thereof.

[0068] (Appendix 13) 13. The chromatography composition of claim 12, wherein I is a tertiary amine.

[0069] (Appendix 14) 11. The chromatographic composition of claim 10, wherein I is an immobilized carboxylic or sulfonic acid.

[0070] (Appendix 15) The ionically modified hydrophilic ligand has formula IV: [ka] 14. The chromatography composition of any one of claims 1 to 13, derived from

[0071] (Appendix 16) 16. The chromatography composition of any one of claims 1 to 15, further comprising a hydrophilic ligand covalently attached to the solid phase substrate, the hydrophilic ligand comprising a polar group and multiple hydroxyl groups.

[0072] (Appendix 17) The hydrophilic ligand has formula V: [ka] (where: X is a polar group, n is 1-6, n' is 0-2, m is 2-8, q is 1, R 1 , R 2, R 3 is independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group; R 4 and R 5 is independently H or OH, and at least two m units contain at least one hydroxyl group; and R 8 and R 9 R, provided that at least one of 8 and R 9 are independently H or OH. 17. The chromatography composition of claim 16, derived from

[0073] (Appendix 18) The hydrophilic ligand of formula V has formula Va: [ka] 18. The chromatography composition of claim 17, derived from

[0074] (Appendix 19) 19. The chromatography composition of any one of claims 16 to 18, wherein the ionically modified hydrophilic ligand and the hydrophilic ligand are present in a molar ratio range of 1:10 to 10:1.

[0075] (Appendix 20) 20. The chromatography composition of any one of claims 1 to 19, wherein the solid phase substrate is a silica material or an inorganic / organic hybrid material.

[0076] (Appendix 21) The ionically modified hydrophilic ligand attached to the solid substrate comprises [ka] 21. The chromatography composition according to any one of claims 1 to 20, wherein

[0077] (Appendix 22) 22. A chromatography composition according to any one of claims 1 to 21 for use in hydrophilic interaction liquid chromatography or mixed-mode hydrophilic interaction liquid chromatography.

[0078] (Appendix 23) 22. A kit comprising the chromatography composition of any one of claims 1 to 21.

[0079] (Appendix 24) 22. A separation device comprising the chromatography composition of any one of claims 1 to 21, further defined as a chromatography column, a thin layer plate, a filtration membrane, a microfluidic separation device, a sample clean-up device, a solid support, a solid phase extraction device, a microchip separation device, or a microtiter plate.

[0080] (Appendix 25) 1. A method for preparing a chromatographic composition comprising an ionically modified hydrophilic ligand, comprising: Providing a solid phase substrate; Providing a hydrophilic ligand comprising a polar group and a plurality of hydroxyl groups, at least one hydroxyl group being present at a terminal end of said hydrophilic ligand; reacting the solid phase substrate with the hydrophilic ligand to covalently bond the hydrophilic ligand to the solid phase substrate to form a hydrophilically modified substrate; Providing an activated compound that includes a leaving group; reacting the activating compound with a terminal hydroxyl group of the hydrophilically modified substrate to form an activated hydrophilically modified substrate; Providing an ionic modifier comprising a nucleophile and an ionic group; and reacting the activated hydrophilically modified substrate with the ionic modifier to release the leaving group of the activated compound and form the ionic modified hydrophilic ligand; The method includes:

[0081] (Appendix 26) The hydrophilic ligand has the formula V: [ka] (where: X is a polar group, n is 1-6, n' is 0-2, m is 2-8, q is 1, R 1 , R 2 , R 3 is independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group; R 4 and R 5 is independently H or OH, and at least two m units contain at least one hydroxyl group; and R 8 and R 9 is R 8 and R 9 is independently H or OH, provided that at least one of is OH and represents the hydroxyl group present at the terminus of the hydrophilic ligand. The method according to claim 25, represented by the formula:

[0082] (Appendix 27) 27. The method of claim 26, wherein the polar group X is independently selected from carbonate, carbamate, amide, amine, urea, ether, thioether, sulfinyl, sulfoxide, sulfonyl, thiourea, thiocarbonate, or thiocarbamate, and comprises a heterocyclic compound containing a polar functional group.

[0083] (Appendix 28) 28. The method of claim 26 or 27, wherein the polar group X is selected from an amide group, a carbamate group, or a ureido group.

[0084] (Appendix 29) 27. The method of claim 26, wherein the polar group X is an amide.

[0085] (Appendix 30) n is 2-4, m is 3-6, p is 1, and R 1 , R 2 , R 3 are independently H or a straight or branched chain, substituted or unsubstituted C1 to C6 alkyl group; 30. The method of any one of claims 26 to 29.

[0086] (Appendix 31) The hydrophilic ligand of formula V is [ka] 31. The method according to any one of claims 24 to 30, wherein

[0087] (Appendix 32) The ionic modifier has formula VI: [ka] (where: W is a nucleophile; k is 1-8, R 6 and R 7 is independently H or a straight or branched chain, substituted or unsubstituted C1 to C18 alkyl group, which may contain halocarbon or alcohol substituents; I, i. carrying a positive ionic charge under neutral or acidic aqueous or aqueous organic solvent conditions; or ii. bears a negative ionic charge under suitable neutral or basic aqueous or aqueous organic solvent conditions; It is a charge-carrying functional group that can 32. The method according to any one of claims 24 to 31, wherein

[0088] (Appendix 33) 33. The method of any one of claims 24 to 32, wherein the ionic modifying agent is selected from N,N-(diethyl)-diaminoethane, 4-(aminoethyl)pyridine, 2-aminoethanesulfonic acid, 2-aminoethanesulfinic acid, and 3-aminopropanesulfonic acid.

[0089] (Appendix 34) 33. The method of claim 32, wherein the ionic modifying agent is N,N-(diethyl)-diaminoethane.

[0090] (Appendix 35) 33. The method of any one of claims 24 to 32, wherein the ionic group of the ionic modifier is a tertiary amine.

[0091] (Appendix 36) 36. The method of any one of claims 24 to 35, wherein the activating compound comprises a carbonate and is represented by 4-nitrophenyl chloroformate (4-NPC), N,N'-disuccinimidyl carbonate (DSC), carbonyldiimidazole (CDI), or a combination thereof.

[0092] (Appendix 37) 36. The method of any one of claims 24 to 35, wherein the activated compound comprises a tosylate group.

[0093] (Appendix 38) 38. The method of claim 37, wherein the activating compound is tosyl chloride.

[0094] (Appendix 39) 35. The method of any one of claims 24 to 34, wherein the activating compound is mesyl chloride, phosphorus tribromide, thionyl chloride, or a combination thereof.

[0095] (Appendix 40) The ionically modified hydrophilic ligand has formula I: [ka] (where: X is a polar group, Z is a polar linking group; Y is an ionic group; n is 1-6, n' is 0-2, m is 2-8, p is 0 or 1, s is 1, R 1 , R 2 , R 3 are independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group, and R 4 and R 5 is independently H or OH, and at least two m units contain at least one hydroxyl group. 40. The method according to any one of claims 24 to 39, wherein

[0096] (Appendix 41) The ionically modified hydrophilic ligand is [ka] 41. The method according to any one of claims 24 to 40, wherein

[0097] (Appendix 42) 1. A method for preparing a chromatographic composition comprising an ionically modified hydrophilic ligand, comprising: Providing a solid phase substrate; Providing a hydrophilic ligand comprising a polar group and a plurality of hydroxyl groups, at least one hydroxyl group being present at a terminal end of said hydrophilic ligand; reacting the solid phase substrate with the hydrophilic ligand to covalently bond the hydrophilic ligand to the solid phase substrate to form a hydrophilically modified substrate; Providing an activated compound that includes a leaving group; Providing an ionic modifier comprising a nucleophile and an ionic group; reacting the activating compound with the ion modifier to form an activating ion producing compound; and reacting a terminal hydroxyl group of said hydrophilically modified substrate with said activating ion generating compound to form said ionically modified hydrophilic ligand; The method includes:

[0098] (Appendix 43) 43. The method of claim 42, wherein the activated compound comprises a first leaving group and a second leaving group.

[0099] (Appendix 44) 44. The method of claim 43, wherein the first leaving group of the activated compound is released during reaction of the activated compound with the ion modifier and the second leaving group is released during reaction of the hydrophilic modified substrate with the ion-producing compound.

[0100] (Appendix 45) The hydrophilic ligand has the formula V [ka] (where: X is a polar group, n is 1-6, n' is 0-2, m is 2-8, q is 1, R 1 , R 2 , R 3 is independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group; R 4 and R 5 is independently H or OH, and at least two m units contain at least one hydroxyl group; and R 8 and R 9 is R 8 and R 9 are independently H or OH, provided that at least one of is OH, representing the hydroxyl group present at the terminus of the hydrophilic ligand. The method according to claim 42, represented by the formula:

[0101] (Appendix 46) 46. ​​The method of claim 45, wherein the polar group X is an amide.

[0102] (Appendix 47) n is 2-4, m is 3-6, p is 1, and R 1 , R 2 , R 3 are independently H or a straight or branched chain, substituted or unsubstituted C1 to C6 alkyl group; 47. The method of any one of claims 42 to 46.

[0103] (Appendix 48) The hydrophilic ligand of formula V is [ka] 48. The method according to any one of claims 42 to 47, wherein

[0104] (Appendix 49) 49. The method of any one of claims 42 to 48, wherein the ionic modifier is N,N-(diethyl)-diaminoethane.

[0105] (Appendix 50) 50. The method of any one of claims 42 to 49, wherein the activated compound contains a carbonate group and is represented by 4-nitrophenyl chloroformate (4-NPC), N,N'-disuccinimidyl carbonate (DSC), carbonyldiimidazole (CDI), or a combination thereof.

[0106] (Appendix 51) The ionically modified hydrophilic ligand is [ka] 51. The method of any one of claims 42 to 50, wherein

Claims

1. A solid phase substrate, and the solid phase substrate comprising an ionically modified hydrophilic ligand attached thereto, the ionically modified hydrophilic ligand comprising a hydrophilic ligand moiety covalently attached to said solid phase substrate, said ligand moiety comprising a polar group and a plurality of hydroxyl groups; and an ionic group attached directly or indirectly to said hydrophilic ligand moiety; Including, Chromatographic compositions.

2. The ionically modified hydrophilic ligand has formula I: 【Chemistry 1】 (where: X is a polar group; Z is a polar linking group; Y is an ionic group; n is 1-6; n′ is 0-2; m is 2-8; p is 0 or 1; s is 1, R １ , R ２ , R ３ is independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group; and R ４ and R ５ is independently H or OH, and at least two m units contain at least one hydroxyl group.

2. The chromatography composition of claim 1 , derived from

3. The hydrophilic ligand moiety has the formula Ia: 【Chemistry 2】 (where: X is a polar group; n is 1-6; n′ is 0-2; R １ , R ２ , R ３ is independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group; and R ４ and R ５ is independently H or OH, and at least two m units contain at least one hydroxyl group.

3. The chromatography composition of claim 1 or 2, which is derived from

4. 4. The chromatographic composition of claim 1, wherein the polar group X is independently selected from carbonate, carbamate, amide, amine, ureido, ether, thioether, sulfinyl, sulfoxide, sulfonyl, thiourea, thiocarbonate, or thiocarbamate and comprises a heterocyclic compound containing a polar functional group.

5. 5. The chromatographic composition of claim 4, wherein the polar group X is selected from an amide, a carbamate, or a ureido group.

6. The chromatographic composition of claim 4 , wherein the polar group X is an amide.

7. n is 2-4; m is 3-6; p is 1, and R １ , R ２ , R ３ is independently H or a straight or branched, substituted or unsubstituted C1 to C6 alkyl group; 7. The chromatography composition according to any one of claims 1 to 6.

8. 8. The chromatographic composition of claim 1, wherein the polar linking group Z is a carbamate group and p is 1.

9. The ionically modified hydrophilic ligand has the formula II: 【Chemistry 3】 9. The chromatography composition of claim 1 , which is derived from

10. The ionic group Y is represented by formula III: 【Chemistry 4】 (where: k is 1-8; R ６ and R ７ is independently H or a straight or branched chain, substituted or unsubstituted C1 to C18 alkyl group, which may also contain halocarbon or alcohol substituents; I is a charged functional group, i. bears a positive ionic charge under neutral or acidic aqueous or aqueous organic solvent conditions; or ii. Carry a negative ionic charge under suitable neutral or basic aqueous or aqueous organic solvent conditions; It is possible.) The chromatography composition according to any one of claims 1 to 9, wherein

11. 11. The chromatographic composition of claim 10, wherein I is a primary, secondary, tertiary or quaternary amine.

12. 12. The chromatography composition of claim 11, wherein the nitrogen atom of the amine is bonded to a hydrogen, an alkyl, an alcohol substituted alkyl, an aromatic group, and combinations thereof.

13. 13. The chromatographic composition of claim 12, wherein I is a tertiary amine.

14. 11. The chromatographic composition of claim 10, wherein I is an immobilized carboxylic or sulfonic acid.

15. The ionically modified hydrophilic ligand has formula IV: 【Chemistry 5】 14. The chromatography composition of claim 1 , derived from

16. 16. The chromatography composition of claim 1, further comprising a hydrophilic ligand covalently attached to the solid phase substrate, the hydrophilic ligand comprising a polar group and multiple hydroxyl groups.

17. The hydrophilic ligand has formula V: 【Chemistry 6】 (where: X is a polar group; n is 1-6; n′ is 0-2; m is 2-8; q is 1; R １ , R ２ , R ３ is independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group; R ４ and R ５ is independently H or OH, and at least two m units contain at least one hydroxyl group; and R ８ and R ９ R ８ and R ９ are independently H or OH.

17. The chromatography composition of claim 16, derived from

18. The hydrophilic ligand of formula V has formula Va: 【Chemistry 7】 20. The chromatography composition of claim 17, derived from

19. 19. The chromatography composition of any one of claims 16 to 18, wherein the ionically modified hydrophilic ligand and the hydrophilic ligand are present in a molar ratio range of from 1:10 to 10:

1.

20. 20. The chromatographic composition of claim 1, wherein the solid phase substrate is a silica material or an inorganic / organic hybrid material.

21. The ionically modified hydrophilic ligand attached to the solid substrate comprises 【Chemistry 8】 The chromatography composition according to any one of claims 1 to 20, wherein

22. 22. A chromatography composition according to any one of claims 1 to 21 for use in hydrophilic interaction liquid chromatography or mixed-mode hydrophilic interaction liquid chromatography.

23. 22. A kit comprising the chromatography composition of any one of claims 1 to 21.

24. 22. A separation device comprising the chromatography composition of any one of claims 1 to 21 and further defined as a chromatography column, a thin layer plate, a filtration membrane, a microfluidic separation device, a sample clean-up device, a solid support, a solid phase extraction device, a microchip separation device, or a microtiter plate.

25. 1. A method for preparing a chromatographic composition comprising an ionically modified hydrophilic ligand, comprising: Providing a solid phase substrate; Providing a hydrophilic ligand comprising a polar group and a plurality of hydroxyl groups, at least one hydroxyl group being present at a terminal end of said hydrophilic ligand; reacting the solid phase substrate with the hydrophilic ligand to covalently bond the hydrophilic ligand to the solid phase substrate to form a hydrophilically modified substrate; Providing an activated compound that includes a leaving group; reacting the activating compound with a terminal hydroxyl group of the hydrophilically modified substrate to form an activated hydrophilically modified substrate; Providing an ionic modifier comprising a nucleophile and an ionic group; and reacting the activated hydrophilically modified substrate with the ionic modifier to release the leaving group of the activated compound and form the ionic modified hydrophilic ligand; The method includes:

26. The hydrophilic ligand has the formula V: 【Chemistry 9】 (where: X is a polar group; n is 1-6; n′ is 0-2; m is 2-8; q is 1; R １ , R ２ , R ３ is independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group; R ４ and R ５ is independently H or OH, and at least two m units contain at least one hydroxyl group; and R ８ and R ９ is R ８ and R ９ are independently H or OH, provided that at least one of is OH and represents the hydroxyl group present at the terminus of the hydrophilic ligand. The method of claim 25 , wherein

27. 27. The method of claim 26, wherein the polar group X is independently selected from carbonate, carbamate, amide, amine, urea, ether, thioether, sulfinyl, sulfoxide, sulfonyl, thiourea, thiocarbonate, or thiocarbamate, and comprises a heterocyclic compound containing a polar functional group.

28. 28. The method of claim 26 or 27, wherein the polar group X is selected from an amide group, a carbamate group, or a ureido group.

29. 27. The method of claim 26, wherein the polar group X is an amide.

30. n is 2-4; m is 3-6; p is 1, and R １ , R ２ , R ３ is independently H or a straight or branched, substituted or unsubstituted C1 to C6 alkyl group; 30. The method of any one of claims 26 to 29.

31. The hydrophilic ligand of formula V is 【Chemistry 10】 The method according to any one of claims 24 to 30, wherein

32. The ionic modifier has formula VI: 【Chemistry 11】 (where: W is a nucleophile; k is 1-8; R ６ and R ７ is independently H or a straight or branched chain, substituted or unsubstituted C1 to C18 alkyl group, which may contain halocarbon or alcohol substituents; I, i. carrying a positive ionic charge under neutral or acidic aqueous or aqueous organic solvent conditions; or ii. Carrying a negative ionic charge under suitable neutral or basic aqueous or aqueous organic solvent conditions; It is a charge-carrying functional group that can 32. The method according to any one of claims 24 to 31, wherein

33. 33. The method of any one of claims 24 to 32, wherein the ionic modifier is selected from N,N-(diethyl)-diaminoethane, 4-(aminoethyl)pyridine, 2-aminoethanesulfonic acid, 2-aminoethanesulfinic acid, and 3-aminopropanesulfonic acid.

34. 33. The method of claim 32, wherein the ionic modifier is N,N-(diethyl)-diaminoethane.

35. 33. The method of any one of claims 24 to 32, wherein the ionic group of the ionic modifier is a tertiary amine.

36. 36. The method of any one of claims 24 to 35, wherein the activating compound comprises a carbonate and is represented by 4-nitrophenyl chloroformate (4-NPC), N,N'-disuccinimidyl carbonate (DSC), carbonyldiimidazole (CDI), or a combination thereof.

37. 36. The method of any one of claims 24 to 35, wherein the activating compound comprises a tosylate group.

38. 38. The method of claim 37, wherein the activating compound is tosyl chloride.

39. 35. The method of any one of claims 24 to 34, wherein the activating compound is mesyl chloride, phosphorus tribromide, thionyl chloride, or a combination thereof.

40. The ionically modified hydrophilic ligand has formula I: 【Chemistry 12】 (where: X is a polar group; Z is a polar linking group; Y is an ionic group; n is 1-6; n′ is 0-2; m is 2-8; p is 0 or 1; s is 1, R １ , R ２ , R ３ is independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group; and R ４ and R ５ is independently H or OH, and at least two m units contain at least one hydroxyl group.

40. The method according to any one of claims 24 to 39, wherein

41. The ionically modified hydrophilic ligand is 【Chemistry 13】 41. The method according to any one of claims 24 to 40, wherein

42. 1. A method for preparing a chromatographic composition comprising an ionically modified hydrophilic ligand, comprising: Providing a solid phase substrate; Providing a hydrophilic ligand comprising a polar group and a plurality of hydroxyl groups, at least one hydroxyl group being present at a terminal end of said hydrophilic ligand; reacting the solid phase substrate with the hydrophilic ligand to covalently bond the hydrophilic ligand to the solid phase substrate to form a hydrophilically modified substrate; Providing an activated compound that includes a leaving group; Providing an ionic modifier comprising a nucleophile and an ionic group; reacting the activating compound with the ion modifier to form an activating ion producing compound; and reacting a terminal hydroxyl group of said hydrophilically modified substrate with said activator ion generating compound to form said ionically modified hydrophilic ligand; The method includes:

43. 43. The method of claim 42, wherein the activating compound comprises a first leaving group and a second leaving group.

44. 44. The method of claim 43, wherein the first leaving group of the activated compound is released during reaction of the activated compound with the ion modifier and the second leaving group is released during reaction of the hydrophilic modified substrate with the ion generating compound.

45. The hydrophilic ligand has the formula V 【Chemistry 14】 (where: X is a polar group; n is 1-6; n′ is 0-2; m is 2-8; q is 1; R １ , R ２ , R ３ is independently H or a straight or branched, substituted or unsubstituted C1 to C18 alkyl group; R ４ and R ５ is independently H or OH, and at least two m units contain at least one hydroxyl group; and R ８ and R ９ is R ８ and R ９ are independently H or OH, provided that at least one of is OH, representing the hydroxyl group present at the terminus of the hydrophilic ligand. The method of claim 42, wherein

46. 46. ​​The method of claim 45, wherein the polar group X is an amide.

47. n is 2-4; m is 3-6; p is 1, and R １ , R ２ , R ３ is independently H or a straight or branched, substituted or unsubstituted C1 to C6 alkyl group; 47. The method of any one of claims 42 to 46.

48. The hydrophilic ligand of formula V is 【Chemistry 15】 48. The method according to any one of claims 42 to 47, wherein

49. 49. The method of any one of claims 42 to 48, wherein the ionic modifier is N,N-(diethyl)-diaminoethane.

50. 50. The method of any one of claims 42 to 49, wherein the activating compound comprises a carbonate group and is represented by 4-nitrophenyl chloroformate (4-NPC), N,N'-disuccinimidyl carbonate (DSC), carbonyldiimidazole (CDI), or a combination thereof.

51. The ionically modified hydrophilic ligand is 【Chemistry 16】 51. The method according to any one of claims 42 to 50, wherein

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