JPH0943222A - Filler for liquid chromatography and its producing - Google Patents

Abstract

PROBLEM TO BE SOLVED: To efficiently separate hardly separable compounds, especially polychlorinated biphenyl(PCB) group and Fullerene group by constituting with a carrier chemically modified with polycycic aromatic functional group with a specific fused ring number. SOLUTION: A silylation reagent having a polycyclic aromatic functional group with a specific fused ring number of 5 to 10 is reacted with hydroxyl group on the carrier surface and a stationary phase having a polycyclic aromatic functional group is formed on the carrier surface. For a polycyclic aromatic functional group with a specific fused ring number of 5 to 10, a picenyl group or perylenyl group, etc., are raised for examples. In the case of ring number of 5 or less, sufficient tenacity is not obtained and the separation of the objective compounds becomes hard. In the case of 10 or more, bonding quantity of the stationary phase to the carrier decreases and tenacity lowers. Therefore, it is disable to bond with the carrier as much as possible to gain larger tenacity of PCB group and Fullerene group. For the carrier, silica gel is favorable because the adoption of this makes the surface area of carrier material large and the amount of bonding stationary phase becomes large and so the separation capability of the objective compound improves.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel packing material for liquid chromatography and a method for producing the same, and particularly for liquid chromatography having a structure capable of effectively increasing the retention of polychlorinated biphenyls and fullerenes. A filler and a method for producing it advantageously. The present invention also relates to a method for separating polychlorinated biphenyls or fullerenes using such a packing material for liquid chromatography.

[0002]

2. Description of the Related Art Conventionally, a liquid chromatography technique can apply a wide range of separation conditions, does not require exposing a sample to high temperatures,
Since compounds can be separated, they have been widely used for purification and separation of various compounds ranging from biological substances to organic compounds in general. Since the separation ability of substances by this liquid chromatography method is greatly influenced by the performance of the packing material packed in the column, it is possible to enhance the separation ability of compounds by the liquid chromatography method by selecting the packing material. Can be done.

By the way, in general, as such a packing material for liquid chromatography, a C ₁₈ type packing material having an octadecyl group bonded to a carrier is often used. And
The column using this C ₁₈ type packing material (hereinafter referred to as C ₁₈ type column) has its holding power exerted by the interaction between the hydrophobicity of the C ₁₈ type packing material and the hydrophobicity of the sample. However, recently, since various compounds have been separated by a liquid chromatography technique, the number of compounds that cannot be sufficiently separated by such a C ₁₈ column is increasing.

One of the compounds that is difficult to separate on such a C ₁₈ column is, for example, polychlorinated biphenyls (hereinafter referred to as PCB). These PCBs are used in various fields because they are excellent in flame resistance, thermal stability, oxidation resistance, electrical characteristics, solubility, inertness, liquidity, etc. Since a large amount of PCBs have been used as electric components such as capacitors and heat mediums, much more PCBs have been released into the environment than dioxins, which have recently been attracting attention as environmental problems. Thus, in recent years, toxicity of PCBs has become clear, and environmental pollution by PCBs has become a serious problem, and analysis of PCBs, particularly analysis of toxicity has become necessary.

However, PCBs have more than 200 kinds of isomers, and it is difficult to separate these isomers, and the number of chlorine atoms bonded to the biphenyl skeleton and the bond are Due to the different positions, different toxicity is exerted, which makes it difficult to perform toxicity analysis.

[0006] That is, PCBs generally have strong toxicity when the biphenyl skeleton does not have a chlorine atom bonded to the 2nd or 6th position, where the positions bonded to each other are the 1st position. It works, especially 3,
3 ', 4,4'-tetrachlorobiphenyl (IUPAC
No. 77), 3,3 ', 4,4', 5-pentachlorobiphenyl (IUPAC No. 126), and 3,
It has been clarified that 3 ', 4,4', 5,5'-hexachlorobiphenyl (IUPAC No. 169) is highly toxic. Further, when evaluating the toxicity of various PCB samples, it is not possible to accurately evaluate the toxicity only by measuring the total amount of PCB, and therefore the highly toxic PCB
It is necessary to separate only the species and measure the amount.

However, for example, when quantifying highly toxic PCBs using a gas chromatography technique, an activated carbon column is generally used to pretreat the PCBs. Since the activated carbon column has a low ability to separate highly toxic PCBs and a plurality of activated carbon columns are required to sufficiently separate PCBs, the efficiency in quantifying PCBs becomes poor. is there. Further, in the case of directly analyzing PCBs using the capillary GC-MS, there arises a problem that sufficient sensitivity cannot be obtained because the sample load is small. Furthermore,
PY in which pyrenylethyl group is bonded to silica gel as a carrier
In the case of separating highly toxic PCBs by liquid chromatography using a column containing an E packing (hereinafter referred to as PYE column), the retention time of PCBs by such PYE column is short and sufficient. The current situation is that no separation can take place. Therefore, from such a background, it is desired to develop a method for efficiently separating highly toxic PCBs.

Fullerene is also one of the compounds that cannot be efficiently separated by a C ₁₈ column. These fullerenes are typified by C ₆₀ , C _70, etc.
A simple substance composed of only carbon atoms, or a compound group in which various functional groups are bound to this simple substance as a basic skeleton, and is used in fields such as superconducting materials, ferromagnetic materials, pharmaceuticals, lubricating oils, and semiconductors. It is a new substance that is expected to have new functions that can be applied.

By the way, these fullerenes are produced together with soot generated during arc discharge or resistance heating of a graphite rod and are separated from the soot by extracting the soot with an aromatic hydrocarbon solvent such as toluene. ing.
Therefore, at present, since the technology for synthesizing only a single type of fullerene has not been established, the allotropes of various fullerenes are contained in the toluene extraction solution of the soot. It is necessary to separate and purify each allotrope in order to exert the useful function. However, fullerenes are very high in allotropes or isomers having similar properties, are high melting point substances, and may change to other allotropes under high temperature conditions. Therefore, it is difficult to isolate the compound by a purification method such as distillation, recrystallization, sublimation, etc., which is often used for the purification of ordinary compounds. Therefore, for the separation and purification of such fullerenes, the separation ability is excellent, and it is possible to separate the fullerenes under mild conditions,
Separation using liquid chromatography techniques is expected.

However, even with this fullerene, in a solvent such as toluene which dissolves fullerenes well, C
There was a problem that it could not be separated well because it was not retained on the ₁₈ series column.

Therefore, when such a C ₁₈ column or other existing packing material for liquid chromatography is used, there are some difficult-to-separate compounds that cannot be sufficiently separated. There is a strong demand for the development of a packing material for liquid chromatography capable of efficiently separating such difficult-to-separate compounds, particularly PCBs and fullerenes.

[0012]

In order to solve such a problem, the present inventor has conducted an earnest search for a functional group having an excellent coercive force, and as a result, the carrier is chemically treated with a polycyclic aromatic functional group. The filler obtained by the modification shows a stronger holding power for highly toxic PCBs, and the holding power increases as the number of condensed rings constituting the polycyclic aromatic functional group increases. I found the property of becoming stronger. As a result of applying such a filler to the separation of PCBs, it is possible to effectively separate only highly toxic PCBs that were difficult to separate when the conventional filler was applied. It has become possible. Further, when such a polycyclic aromatic functional group-bonded filler is applied to the separation of fullerenes, a filler that recognizes a difference in the number of carbon atoms that advantageously constitutes fullerenes better than conventional fillers It was confirmed that it was possible to efficiently separate the fullerenes by using the filler according to the present invention.

That is, in the present invention, the number of condensed rings is 5 to 1
The gist of a packing material for liquid chromatography is characterized in that it is composed of a carrier chemically modified with a polycyclic aromatic functional group of 0.

Therefore, in such a packing material for liquid chromatography according to the present invention, since the carrier is chemically modified with the stationary phase having a polycyclic aromatic functional group, various kinds of carriers are obtained due to the hydrophobicity of the stationary phase. The compounds are retained by inducing a hydrophobic interaction with the compounds. Also,
In addition, since the polycyclic aromatic functional group is an electron-donating functional group having many π electrons, it has an electron-withdrawing substituent, that is, is induced between the compound and the electron-accepting compound. The resulting charge transfer interaction is increased, so that the compound having an electron withdrawing group is more strongly retained. Further, the π electron of the polycyclic aromatic functional group has a large electronic interaction with the π electron of the compound having a π electron,
The compound having an electron is also strongly retained.

According to a preferred embodiment of the packing material for liquid chromatography according to the present invention, silica gel is used as the carrier.

The use of silica gel as the carrier increases the surface area of the carrier material, and the amount of the stationary phase bound to the carrier surface can be easily increased. The holding power, and eventually the separability, can be further improved.

In the present invention, a silylating agent having a polycyclic aromatic functional group having 5 to 10 condensed rings is used, and the silylating agent is reacted with a hydroxyl group on the surface of the carrier to give A gist also includes a method for producing a packing material for liquid chromatography, which comprises forming a stationary phase having a ring aromatic functional group on the surface of the carrier.

That is, as described above, the number of condensed rings is 5 to 5.
A stationary phase having a polycyclic aromatic functional group of 10 is prepared as a silylating agent, and by reacting it with a hydroxyl group on the surface of the carrier, the stationary phase is efficiently and reliably introduced into the carrier. You will get it.

Furthermore, in a preferred embodiment of the method for producing a packing material for liquid chromatography according to the present invention, the carrier is silica gel as described above.

By using silica gel as the carrier, the surface area of the carrier material is increased, and the reactivity of the silylating agent with the hydroxyl groups on the carrier surface is improved. A filler having a large amount, in other words, an increased holding power, can be more advantageously obtained. Further, since the reactivity of the silylating agent with the carrier is increased, even if the amount of the silylating agent reacted with the carrier is reduced, it is possible to obtain a filler having excellent holding power. Become.

Furthermore, according to the present invention, when separating polychlorinated biphenyls or fullerenes by a liquid chromatography method, a carrier chemically modified with a polycyclic aromatic functional group having 5 to 10 condensed rings is used. A separation method characterized by using a constituted filler is also the subject of the invention.

That is, in this separation method, since the filler composed of the carrier chemically modified with the polycyclic aromatic functional group having 5 to 10 condensed rings is used, Since the aromatic functional group and the highly toxic polychlorinated biphenyls are likely to interact with each other, the highly toxic polychlorinated biphenyls and the less toxic polychlorinated biphenyls are advantageously separated. Further, since the interaction between the polycyclic aromatic functional group and the fullerenes differs depending on the difference in the number of carbon atoms constituting the fullerenes, the fullerene allotrope is also advantageously separated. As described above, in the present invention, when a conventional filler is used, it is possible to effectively separate isomers and allotropes that were difficult to separate due to insufficient holding power. To get.

[0023]

BEST MODE FOR CARRYING OUT THE INVENTION By the way, as the filler according to the present invention, if the carrier is chemically modified with a polycyclic aromatic functional group having 5 to 10 condensed rings as described above, It doesn't matter what form it is,
Generally, it is a chemically bonded filler represented by an Ar-A-O-support material (provided that Ar: polycyclic aromatic functional group, A: spacer group, O: oxygen atom).

The polycyclic aromatic functional group is chemically bonded to the polycyclic aromatic functional group through a predetermined spacer group (A) to give a desired filler. Any known carrier material can be used as long as it can be physically bound and retained. In producing such a filler, an organosilicon compound (well known as a general method for producing a filler) is used. In the case of adopting a chemical modification method using a silylating agent), porous glass, glass beads, alumina, titania, zirconia, etc. are appropriately used in addition to silica gel. However, considering the surface area of the carrier material and the reactivity of the silylating agent with the hydroxyl groups on the surface of the carrier, silica gel or porous glass is preferably used among the above carriers, and silica gel is more preferably used.

The silylating agent reacts not only with the hydroxyl groups present on the surface of the inorganic carrier material, but also with the alcoholic hydroxyl groups present on the organic carrier. The reaction between the silylating agent and the alcoholic hydroxyl group forms an alkoxysilane (Si-O-R), and the bond is easily hydrolyzed, so that the water-alcohol mixed solvent is used as the mobile phase solvent. It can not be used under various conditions, but when liquid chromatography is performed under conditions that do not use water or alcohol as the mobile phase solvent, polystyrene gel,
It is also possible to use an organic polymer gel such as polyvinyl alcohol gel or a carrier material based on a polysaccharide such as agarose or cellulose.

Further, as the spacer group represented by A above, which is bonded between the polycyclic aromatic functional group and the carrier material, most of the holding power of the filler for PCBs and fullerenes is Various known atomic groups can be used because they are expressed in the polycyclic aromatic functional group portion and are hardly affected by the difference in the spacer group. And
Examples of such a spacer group include a straight chain hydrocarbon group such as a methylene group, an ethylene group and a propylene group, and in addition to these, an ether bond, an amide bond, an ester bond, a carbonyl group and the like, and a sulfur atom. It does not matter even if an atomic group into which a functional group containing a nitrogen atom is introduced is used. Further, here, the structure having a spacer group is exemplified, but the structure is not limited thereto, and the polycyclic aromatic functional group is directly bonded to the carrier material without a spacer group. However, there is no problem.

In the present invention, the chemical modification of the carrier material is carried out with a polycyclic aromatic functional group having 5 to 10 fused rings. However, if the number of condensed rings of the polycyclic aromatic functional group is less than 5, sufficient holding power cannot be obtained, and it becomes difficult to separate the target compound. When the number of condensed rings of the group functional group is more than 10, the polycyclic aromatic functional group becomes large, and the amount of the stationary phase bound to the carrier decreases due to the reaction inhibition due to steric hindrance, resulting in the retention force. Is reduced,
Furthermore, as the number of condensed rings increases, the naturally occurring amount decreases, and it becomes difficult to obtain the necessary amount of polycyclic aromatic compound when manufacturing the filler.

As such a polycyclic aromatic functional group, any polycyclic aromatic functional group having a condensed ring number of 5 to 10 can be adopted, for example, a picenyl group and a perylenyl group. , Pentaphenyl group, pentacenyl group, tetraphenylene group, benzofluoranthinyl group,
Hexaphenyl group, hexacenyl group, rubicenyl group, coronenyl group, trinaphthylenyl group, heptaphenyl group,
Examples thereof include a heptaenyl group, a pyranthrenyl group, a decacyclenyl group, and an ovarenyl group. Further, these polycyclic aromatic functional groups are preferably composed of only a 6-membered ring because the strain of the ring is reduced and the conjugation of π electrons on the ring is not hindered. Within a range that does not affect the performance as, for example, including a 5-membered ring in the constituent ring such as the benzofluoranthinyl group or decacyclenyl group, or an 8-membered ring such as the tetraphenylene group. May be included in. Further, the polycyclic aromatic functional group may have a substituent on its ring without any problem.

By the way, the amount of the stationary phase (polycyclic aromatic functional group) in the filler according to the present invention affects the retention time of the solute, but generally, when the amount increases, the solute and the stationary phase are increased. Interaction with the solute, resulting in increased solute retention. This relationship can be similarly applied to PCBs and fullerenes, and the use of a filler composed of a large number of stationary phases bonded together exerts a strong retaining force on PCBs and fullerenes. The separation of these compounds can be effectively performed. That is, it is desirable that as many polycyclic aromatic functional groups as possible be bound to the carrier in order to obtain a filler having a greater holding power for PCBs and fullerenes.

On the other hand, the lower limit of the amount of the stationary phase in the filler according to the present invention varies depending on the conditions and is not unconditionally determined, but generally, the retention force for the target substance is effectively exhibited. It is enough if there is a sufficient amount. Therefore,
When the carrier is chemically modified with a stationary phase having a polycyclic aromatic functional group, even if the reactivity thereof is slightly worse than the reactivity of the conventional filler, it is adopted in the filler according to the present invention. Since the stationary phase has a strong holding power for solutes, it is possible to obtain a filler that can be sufficiently used. That is, when chemically modifying with a polycyclic aromatic functional group,
Even if the carrier has a poor reactivity with the polycyclic aromatic functional group, a more useful filler than the conventional filler may be obtained. Therefore, the range of choice of the carrier to be reacted is increased. Can be advantageously spread. Specifically, for example, in the case of a filler having three times the holding power as compared with the conventional filler, even if the amount of the stationary phase of the filler becomes 1/3, it is almost the same as that of the conventional filler. Has the holding power of
If the amount of the stationary phase is made larger than that, it becomes a useful filler with improved holding power.

Here, the packing material for liquid chromatography according to the present invention can be manufactured by utilizing various known reactions. Generally, 1) a silylating agent having a polycyclic aromatic functional group is used. Method of reacting with carrier 2)
Reaction such as a method of reacting an organometallic compound having a polycyclic aromatic functional group with a carrier, 3) a method of introducing a reactive functional group into a carrier and then bonding a polycyclic aromatic functional group to the functional group Will be manufactured by using.

More specifically, in the above method 1) using a silylating agent, first, a substituent having a double bond is introduced into a polycyclic aromatic functional group. Then, the introduction of the substituent having a double bond into the polycyclic aromatic functional group is specifically performed by introducing a substituent having a double bond such as an allyl group into the polycyclic aromatic functional group, The following reaction formula (1
The reactions shown in a), (1b) and (1c) can be advantageously carried out. Here, Ar represents a polycyclic aromatic functional group and CuX represents a copper halide, and the same applies to the following reaction formulas.

Ar-H + Br ₂ ---> Ar-Br ... (1a) Ar-Br + Mg ----> Ar-MgBr ... (1b) Ar-MgBr + CuX + Br-CH _{2 -CH = CH 2 ─── → Ar-} CH 2 -CH = CH 2 ··· (1c)

The introduction of the substituent having a double bond is not limited to the above reaction and may be carried out, for example, by the following reaction formulas (1d) to (1g). In addition, here, a 4-butenyl group is introduced as a substituent having a double bond.

Ar-H + CH ₃ OCHCl ₂ ---> Ar-CHO (1d) Ar-CHO + BrMgCH ₂ CH = CH ₂ ---- → Ar-CH (OH) CH ₂ CH = CH ₂ ··· (1e) Ar-CH ( OH) CH 2 CH = CH 2 ─── → Ar-CHClCH 2 CH = CH 2 ··· (1f) Ar-CHClCH 2 CH = CH 2 ─── → Ar- _{CH 2 CH 2 CH = CH 2} ··· (1g)

Then, at the double bond site of the polycyclic aromatic functional group having a double bond obtained in the above reaction formula (1c), a hydrosilane is used with a platinum catalyst as shown in the following reaction formula (2). A silylating agent can be obtained by a so-called hydrosilylation reaction in which (H-Si) is added. In this reaction, in the addition reaction of hydrosilane, the anti-Markovnikov type addition mainly proceeds and the silicon atom is bonded to the terminal of the alkene molecule. Such a method has an advantage that a silylating agent having a target structure can be easily obtained because the conversion rate to a silylating agent is large and side reactions are few.

[0037] _{Ar-CH 2 -CH = CH 2} + H-Si (CH 3) 2 -Cl Pt ─── → Ar- (CH 2) 3 -Si (CH 3) 2 -Cl ··· (2)

As the reactive substituent of the silylating agent thus synthesized, there may be mentioned a chlorosilane (Si-Cl) group and the like. Such a reactive substituent can be bonded to a silicon atom. Of the three substituents, the silylating agent is not necessarily limited to only one and has two or three reactive functional groups, and can also be applied to the chemical modification of the carrier. And, as the reactive functional group of such a silylating agent,
Not limited to chlorosilane to silicon atoms of chlorine (-Cl) are thus allowed to bind, bromine to a silicon atom (-Br), methoxy (-OCH _3), ethoxy (-OCH ₂ CH _3), alkylamino group (-NR Any functional group capable of reacting with an active site such as a hydroxyl group existing on the surface of the carrier such as ₂ ) can be applied to the present invention.

As the method for synthesizing the silylating agent, the method using the addition reaction to the double bond of hydrosilane has been exemplified above, but the method is not limited to this, and for example, two or more may be used. The silylating agent can also be synthesized by a method of reacting an organic compound having a reactive functional group (mainly chlorine) with an organometallic reagent such as a Grignard reagent.

Then, the silylating agent having a polycyclic aromatic functional group obtained in the reaction formula (2) is reacted with a carrier such as silica gel (HO-Si) as shown in the reaction formula (3). As a result, the intended filler can be obtained.

Ar- (CH ₂ ) ₃ -Si (CH ₃ ) ₂ -Cl + HO-Si ----> Ar- (CH ₂ ) ₃ -Si (CH ₃ ) ₂ -O-Si ... (3 )

The method 2) using the organometallic compound is the method reported by Locke et al. [DC Locke, J.
J. Schermud and B. Banner, Anal. Chem., 44, 90 (197
2)], and in this method, the following reaction formulas (4a) and (4
The intended filler is obtained through the two-step reaction process shown in b). The reaction represented by the reaction formula (4a) is a reaction of chlorinating a hydroxyl group of a carrier (eg silica gel) using a reagent capable of chlorinating a hydroxyl group such as thionyl chloride. The reaction represented by the formula (4b) is the reaction formula (1a),
This is a reaction in which the Grignard reagent obtained according to (1b) is reacted with the chlorinated carrier synthesized in the following reaction formula (4a) to bond the polycyclic aromatic functional group represented by Ar to the carrier. In addition, in a reaction formula, Si shows a silica gel residue.

Si-OH + SOCl ₂ ---> Si-Cl (4a) Ar-MgBr + Si-Cl ----> Ar-Si (4b)

Further, the method of introducing a polyfunctional aromatic functional group to the functional group after introducing the reactive functional group of the above 3) into the carrier is the organometallic compound shown in 2) above as a carrier. This method is similar to the reaction method, but in this method, as shown in the following reaction formula (5a), a reactive functional group such as an amino group is previously introduced into a carrier (for example, silica gel) and reacted with this amino group. A functional group capable of carrying out the reaction, for example, a polycyclic aromatic functional group having a carboxyl group and a carrier into which the reactive functional group obtained in the following reaction formula (5a) is introduced are shown in the following reaction formula (5b). like,
The desired filler can be obtained by condensing with a condensing agent such as dicyclohexylcarbodiimide. In addition, in this example, the reaction in which an amide bond is finally formed is shown, but other combinations of functional groups forming an ester bond, an ether bond, or the like can be similarly performed. In addition, S in the reaction formula
i-O- represents a silica gel residue.

NH ₂ (CH ₂ ) ₃ Si (OCH ₃ ) ₃ + HO-Si ----> NH ₂ (CH ₂ ) ₃ Si (OCH ₃ ) _2- O-Si ... (5a) Ar-COOH + _{NH 2 (CH 2) 3 Si} (OCH 3) 2 -O-Si ─── → Ar-CONH (CH 2) 3 Si (OCH 3) 2 -O-Si ··· (5b)

As described above, the filler according to the present invention can be produced by various synthetic methods, but it is widely known as a method for chemically modifying an inorganic carrier with an organic compound, and it can be used as an organosilicon compound (silylating agent). It is a useful method that allows various functional groups to be bonded to an inorganic carrier depending on the difference in the bonded functional groups, and since the intended filler can be easily obtained, it can be produced by the method of 1) above. preferable.

The present invention is also characterized by liquid chromatography separation of PCBs and fullerenes using the packing material according to the present invention as described above. Although it has become possible to separate PCBs and fullerenes, the same operations and conditions as conventional ones are adopted for the separation operation of such PCBs and fullerenes by the liquid chromatography method. It will be.

That is, in such a separation operation, PCBs and fullerenes are supplied as a solution dissolved in a predetermined solvent to the column packed with the packing material according to the present invention. Become. by the way,
PCBs dissolve well in hydrocarbons such as hexane and isooctane, and aromatic hydrocarbon solvents such as toluene and benzene, and fullerenes are dissolved in aromatic hydrocarbon solvents such as trichlorobenzene, toluene, benzene, and carbon disulfide. When it is separated and purified because it dissolves well,
Toluene, hexane, etc. are used as the solvent for dissolving PCBs, and aromatic hydrocarbon solvent, carbon disulfide, etc. are used for the solvent for dissolving fullerenes, to prepare a higher concentration sample solution for separation. It is possible and preferable.

The PCBs and fullerenes supplied in the form of a solution into the separation column are retained by a packing material, and subsequently, the PCBs and fullerenes are mixed with a mobile phase solvent which is circulated in the separation column. The species are successively eluted. As for the mobile phase solvent, PCBs do not have the same UV absorption region as PCBs, so that PCBs can be detected more favorably and do not interfere with the interaction between the sample and the stationary phase. It is preferable to use a hydrocarbon solvent such as hexane or isooctane that does not have π electrons in, and for fullerenes,
For example, toluene, hexane, or a mixed solvent of toluene and hexane, preferably carbon disulfide or a mixed solvent of carbon disulfide and toluene is used. Then, PCBs and fullerenes are individually eluted with time from the separation column by the mobile phase solvent, and the eluate flowing out from this separation column is separated with time.
It is possible to separate each isomer or allotrope of PCBs and fullerenes.

[0050]

EXAMPLES In order to clarify the present invention in more detail, some examples of the present invention will be shown below.
It goes without saying that the present invention is not subject to any restrictions by the description of such embodiments. In addition, in addition to the following examples, the present invention, in addition to the above-described specific description, various changes, corrections, and modifications based on the knowledge of those skilled in the art without departing from the spirit of the present invention. It should be understood that improvements can be made.

Example 1 Synthesis of Polycyclic Aromatic Bonding Type Filler ── Polycyclic aromatic bond type silica gel filler is 1) introduction of double bond into polycyclic aromatic compound, 2) The stationary phase (polycyclic aromatic functional group + spacer group) structure shown in Table 1 is obtained through a three-step reaction of a reaction between a polycyclic aromatic compound having a double bond and chlorosilane, 3) a reaction between silica gel and chlorosilane. The fillers of Examples 1 to 3 of the present invention and Comparative Examples 1 to 3 were synthesized. In Table 1, the fillers of Inventive Examples 1 and 2 both have the same stationary phase structure, but when the fillers are synthesized, the reaction rates of the stationary phase and silica gel are different, Different abbreviations are used to distinguish them.

[Table 1]

(A) Synthesis of 5-coronenylpentyldimethylsilylated filler --- Synthesis of coronene aldehyde --- First, coronene: 6.0 g was dissolved in 600 mL of carbon disulfide, and tetrachloride was added. After adding titanium (36 mL), dichloromethyl methyl ether (36 mL) was added dropwise thereto over 5 minutes while stirring under a temperature condition of 0 ° C. After dropping, the mixture was stirred under ice cooling for 30 minutes, and further at room temperature.
The reaction was carried out with stirring for 1 hour. After the reaction, the reaction solution was transferred to a separating funnel containing ice and shaken, and then the organic layer was separated. Then, the residual aqueous layer is further extracted with carbon disulfide, and this is combined with the previously separated organic layer,
After drying over sodium sulfate, the carbon disulfide was distilled off under reduced pressure,
Crude crystals of coronene aldehyde were obtained. The sparse crystals obtained here were extracted with xylene using a Soxhlet extractor to remove solids insoluble in xylene, and recrystallized from xylene to obtain the desired coronene aldehyde (yield: 6.2 g). In addition, confirmation of product is NM
Performed using R.

C ₂₄ H ₁₂ + CH ₃ OCHCl ₂ ─── → C ₂₄ H ₁₁ —CHO

¹ H-NMR (CDCl ₃ ): δ (pp
m) 8.5-8.95 (11H, m, Ar), 10.71
(1H, s, -CHO)

Synthesis of 2- (1-hydroxy-4-pentenyl) coronene Magnesium: 0.73 g and tetrahydrofuran: 20
Place mL in a flask and place it in an argon stream at 4
-Bromo-1-butene: 2.7 g was added to prepare a Grignard reagent. Further, 6.0 g of the coronene aldehyde synthesized above was azeotropically distilled with benzene to remove water and then dispersed in tetrahydrofuran. Then, the Grignard reagent was added dropwise thereto. Then, after the Grignard reagent was dropped, the mixture was reacted for 1 hour while heating under reflux. After such reaction, the obtained reaction liquid was cooled to room temperature, water and phosphoric acid were added thereto,
Extracted with toluene. The obtained toluene layer was dried over sodium sulfate, and then toluene was distilled off under reduced pressure to give crude 2-
(1-Hydroxy-4-pentenyl) coronene was obtained (yield: 6.1 g).

[0056] _{C 24 H 11 -CHO + BrMg (} CH 2) 2 CH = CH 2 ─── → C 24 H 11 -CH (OH) (CH 2) 2 CH = CH 2

Synthesis of 2- (1-chloro-4-pentenyl) coronene ── 11.5 g of the crude 2- (1-hydroxy-4-pentenyl) coronene obtained above was mixed with triphenylphosphine. 10.5 g and carbon tetrachloride: 100 mL were added, and 6
The reaction was carried out at reflux for a period of time. After this reaction, ethanol was added to the reaction solution, and the mixture was refluxed for another hour to decompose excess triphenylphosphine. Then, after distilling off the solvent of the obtained reaction solution, the product was purified by silica gel column chromatography using toluene as a developing solvent, and crude 2-
(1-Chloro-4-pentenyl) coronene was obtained (yield: 7.5 g).

C ₂₄ H ₁₁ —CH (OH) (CH ₂ ) ₂ CH═CH ₂ ─── → C ₂₄ H ₁₁ —CHCl (CH ₂ ) ₂ CH═CH ₂

Synthesis of 1-Coronenyl-4-pentene Crude 2- (1-chloro-4-pentenyl) obtained above
Coronene: 7.4 g, tetrahydrofuran: 100 m
After dispersing in L and adding lithium aluminum hydride (1.5 g) thereto, the mixture was reacted for 5 hours while heating under reflux. After refluxing, it was left at room temperature overnight.
After the reaction, water was added to the reaction solution to hydrolyze the lithium aluminum hydride, the desired compound was extracted with benzene, and the extract was dried with calcium chloride.
The benzene thus obtained was distilled off under reduced pressure and then purified by silica gel column chromatography using toluene: hexane = 1: 1 as a developing solvent to obtain the desired 1-coronenyl-4.
-Pentene was obtained (yield: 5.8 g). The obtained 1-coronenyl-4-pentene was confirmed by NMR.

[0060] _{C 24 H 11 -CHCl (CH 2} ) 2 CH = CH 2 ─── → C 24 H 11 - (CH 2) 3 CH = CH 2

¹ H-NMR (CDCl ₃ ): δ (pp
m) 2.14 (2H, tt, J = 7.7Hz, 7.5Hz,
_{-C H 2 CH 2 -Ar),} 2.35 (2H, dt, J =
_{7.5Hz, 7.0Hz, CH 2 = CH} -C H 2),
3.54 (2H, t, J = 7.7Hz, C H 2 -A
r), 5.09 (1H, dd, J = 10.3 Hz [ci
s], 1.5 Hz [geminal], C H ₂ = CH
-), 5.18 (1H, dd, J = 16.0 Hz [tr
ans], 1.5 Hz [geminal], C H ₂ = C
H-), 6.00 (1H, ddt, J = 7.0 Hz, 1
_{0.3Hz, 16.0Hz, CH 2 = C} H -), 8.3
0-8.88 (11H, m, Ar).

Synthesis of 5-Coronenylpentyldimethylchlorosilane Next, the 1-coronenyl-4-pentene obtained above was reacted with chlorosilane by hydrosilation reaction using platinum as a catalyst. , A silylating agent, 5-coronenylpentyldimethylchlorosilane was synthesized.

That is, first, 5 g of 1-coronenyl-4-pentene was dissolved in 100 mL of benzene, and chloroplatinic acid: 20 mg and dimethylchlorosilane: 10 were dissolved in this.
mL was added, and the mixture was reacted at 80 ° C. for 3 hours under reflux. After the reaction, the platinum catalyst is removed by filtration,
Further, excess dimethylchlorosilane and benzene were distilled off under reduced pressure to obtain the desired 5-coronenylpentyldimethylchlorosilane (yield: 5.9 g). The product was confirmed by NMR.

[0064] _{C 24 H 11 - (CH 2} ) 3 CH = CH 2 + H-Si (CH 3) 2 -Cl Pt ─── → C 24 H 11 - (CH 2) 5 -Si (CH 3) 2 -Cl

¹ H-NMR (CDCl ₃ ): δ (pp
m) 0.3 (6H, s, Si-C H 3 × 2), 0.8 (2
_{H, m, Si-CH 2} ), 1.5 (2H, m, -C H 2
_{CH 2 CH 2 -Si), 1.8} (2H, m, -C H 2 C
_{H 2 -Ar), 3.3 (2H} , t, C H 2 -Ar),
8.30-8.88 (11H, m, Ar)

Synthesis of 5-Coronenylpentyldimethylsilylated Filler Next, the 5-coronenylpentyldimethylchlorosilane obtained above was reacted with silica gel as a silylating agent to give 5-corone A nilpentyl silylated filler was obtained.

More specifically, silica gel (particle size: 5 μm
m, pore diameter: 10 nm, surface area: 300 m ² / g): 1
Parts by weight were dispersed in 30 parts by weight of toluene, 2 parts by weight of 5-coronenylpentyldimethylchlorosilane and 0.5 parts by weight of pyridine were added thereto, and the mixture was reacted for 8 hours while heating under reflux. After this reaction, 100 parts by weight of methanol and 100 parts by weight of chloroform were sequentially suction-filtered and washed, and dried at 50 ° C. to obtain the intended filler (COP-5-1) of Inventive Example 1.

The amount of 5-coronenylpentyldimethylchlorosilane to be added was reduced to 1/4 to reduce the reaction rate between the silica gel as the carrier and the silylating agent. The agent (COP-5-2) was similarly obtained.

By the way, in addition to the above reaction, it is possible to synthesize a coronene derivative having a double bond, and a silylating agent is obtained by reacting the obtained coronene derivative with dimethylchlorosilane. Can be done. For example, 2-coronenylethyldimethylchlorosilane can be synthesized from coronene aldehyde via vinyl coronene as follows.

First, 3.6 g of methyltriphenylphosphonium bromide was dispersed in 30 mL of tetrahydrofuran, and then 6.7 mL of 1.5 M n-butyllithium was added dropwise over 5 minutes under room temperature conditions. After the dropping, the mixture was stirred at room temperature for 2 hours. To this, a solution prepared by dispersing 3.3 g of coronene aldehyde synthesized in the same manner as described above in 50 mL of tetrahydrofuran was added, and further reacted at room temperature for 5 hours with stirring. After the reaction, a solution obtained by adding a saturated ammonium chloride aqueous solution to the obtained reaction solution was extracted with ether. Then, the obtained ether layer was dried over sodium sulfate, and the solvent was distilled off under reduced pressure to obtain crude crystals. The crude crystals were dissolved in dichloromethane: 10 mL, and hexane: ethyl acetate =
Purification by silica gel column chromatography using 4: 1 as a developing solvent gave vinyl coronene (yield: 1.
5g).

¹ H-NMR (CDCl ₃ ): δ (pp
m) 5.84 (1H, dd, J = 10.9, 1.57Hz,
= C H ₂ ), 6.24 (1H, dd, J = 17.2,
1.57 Hz, = C H ₂ ), 8.7 (1 H, dd, J =
17.2, 10.9 Hz, CH =), 8.7-9.2
(11H, m, Ar)

Then, in the same manner as in the synthesis of 5-coronenylpentyldimethylchlorosilane from 1-coronenyl-4-pentene, 2-coronenylethyldimethylchlorosilane was synthesized from vinyl coronene (1.5 g) as a raw material ( Yield: 1.9 g). Below, the result of NMR of the obtained product is shown.

¹ H-NMR (CDCl ₃ ): δ (pp
m) 0.29 (6H, s, Si-CH 3 × 2), 1.44
(2H, m, CH ₂ ), 3.74 (2H, m, C
H ₂ ), 8.6-9.0 (11H, m, Ar)

Even with the 2-coronenylethyldimethylchlorosilane thus obtained, the above-mentioned 5-
The desired filler can be obtained by reacting with silica gel in the same manner as in the case of synthesizing the filler of Inventive Example 1 from coronenylpentyldimethylchlorosilane.

(B) Synthesis of 3-perylenylpropyldimethylsilylated filler --- Synthesis of bromoperylene --- First, perylene: 10.0 g was added with carbon tetrachloride: 80 mL, and bromine: 2.0 mL. Was dissolved in 30 mL of carbon tetrachloride, and the solution was added dropwise over 1 hour. Then, after dropping, the mixture was reacted for 6 hours while stirring. After such reaction,
It was left overnight at room temperature. Carbon tetrachloride was distilled off under reduced pressure, ethanol was added and the mixture was refluxed. After cooling, the crude crystals were collected by suction filtration (yield: 11.5 g).

C ₂₀ H ₁₂ + Br ₂ ─── → C ₂₀ H ₁₁ −Br

-Synthesis of allylperylene-Next, in an argon stream, tetrahydrofuran as a solvent was reacted with 0.97 g of magnesium and 11.5 g of bromoperylene obtained in the above reaction, A Grignard reagent was prepared. At this time, in order to allow the reaction to proceed smoothly, methyl iodide was added to start the reaction, and then bromoperylene was added. After the addition was completed, the mixture was reacted under reflux for 1 hour. And after the reaction,
The obtained reaction liquid was cooled to −20 ° C., cuprous iodide (3.8 g) was added, and immediately, allyl bromide (7.3 g) was added dropwise. After the dropping, the reaction was carried out for 2 hours while raising the temperature. After the reaction, it was left overnight. After standing, tetrahydrofuran was distilled off under reduced pressure, a phosphoric acid aqueous solution was added, and the mixture was extracted with toluene and purified by silica gel open column chromatography to obtain the intended allylperylene (yield: 5.1
g). The product was confirmed by NMR.

C ₂₀ H ₁₁ -Br + Mg --- → C ₂₀ H ₁₁ -MgBr C ₂₀ H ₁₁ -MgBr + CuI + Br-CH ₂ -CH = CH ₂ ---- → C ₂₀ H ₁₁ -CH ₂ -CH = CH ₂

¹ H-NMR (CDCl ₃ ): δ (pp
m) 3.75-3.95 (2H, m, C H 2 -Ar), 4.
95-5.45 (2H, m, C H 2 = CH -), 5.9
_{0-6.38 (1H, m, CH 2} = C H -), 7.30
-8.35 (11H, m, Ar)

Then, in the same manner as in the synthesis of 5-coronenylpentyldimethylsilyl, the allyl perylene obtained above was reacted with chlorosilane by a hydrosilation reaction using platinum as a catalyst to perform silylation. 3-Perylenylpropyldimethylchlorosilane, which is an agent, was synthesized.

That is, first, allyl perylene: 5.0 g
Was dissolved in benzene: 100 mL, chloroplatinic acid: 20 mg and dimethylchlorosilane: 10 mL were added, and the mixture was reacted at 80 ° C. for 3 hours under reflux. After the reaction, the platinum catalyst was removed by filtration, and excess dimethylchlorosilane and benzene were distilled off under reduced pressure to obtain the desired 3-perylenylpropyldimethylchlorosilane (yield: 6.1 g). The product can be confirmed by NMR.
I did it in.

¹ H-NMR (CDCl ₃ ): δ (pp
m) 0.25 (6H, s, Si-CH 3 × 2), 1.4 (2
H, m, Si-C H 2), 1.9 (2H, m, Si-C
_{H 2 C H 2), 3.1} (2H, m, C H 2 -Ar),
8.3-7.2 (11H, m, Ar)

Then, the 3-perylenylpropyldimethylchlorosilane obtained above was reacted with silica gel as a silylating agent to obtain a 3-perylenylpropylsilylated filler.

More specifically, silica gel (particle size: 5 μm
m, pore diameter: 10 nm, surface area: 300 m ² / g): 1
Parts by weight are dispersed in toluene: 30 parts by weight, and 3-
After adding 1.7 parts by weight of perylenylpropyldimethylchlorosilane and 0.5 parts by weight of pyridine, the mixture was reacted for 8 hours while heating under reflux. After such a reaction, 100 parts by weight of methanol and 100 parts by weight of chloroform were successively suction-filtered and washed, and dried at 50 ° C.
R-3) was obtained.

The fillers of Comparative Examples 1 to 3 were also synthesized from the corresponding starting compounds in the same manner as the filler of the present invention.

Then, the elemental analysis values of the fillers thus synthesized and the amount of the stationary phase bound to the surface of the silica gel as the carrier were measured, and the results are shown in Table 2 below. Shown in. The silica gel used in this example has a surface area of 300 m ² per gram, and the amount of the stationary phase bound per 1 m ^{2 of} this surface area.
The value shown in (μmol) is the surface reaction rate. The filler having a high surface reaction rate shows that the stationary phase is bound to the surface of the silica gel at a high rate. The surface reaction rate was calculated from the carbon content rate.

[0087]

[Table 2]

Example 2 Comparison of Hydrophobicity and Polarity Discriminating Ability of Packing Agent Each of the packing materials synthesized in Example 1 was packed in a chromatographic tube having an inner diameter of 4.6 mmφ × a length of 150 mm, and various columns were packed. It was made. Then, by using the obtained column, liquid chromatography of various sample solutions was performed to evaluate the hydrophobicity and the polarity discriminating ability of the packing material.

More specifically, the liquid chromatography apparatus includes a liquid feed pump (LC-9A type: manufactured by Shimadzu Corporation), a column packed with each of the packing materials, and a loop type injection valve for injecting a sample. (Made by Rheodyne Co., Ltd.) and an ultraviolet absorption detector (UV detector of SPD-6A type: manufactured by Shimadzu Corporation), and the flow rate of the mobile phase solvent is 1.0 mL / min. And Also,
The detection sensitivity of the ultraviolet absorption detector is 0.08 to 0.16 AU.
The measurement was carried out with the FS set and the detection wavelength set to 254 nm. The column temperature was adjusted with a constant temperature water bath so that the analysis temperature was 30 ° C. A data processor (CR-5A type: manufactured by Shimadzu Corporation) was used to measure the elution time and the peak area.

Further, 60% methanol was used as the mobile phase solvent, and three kinds of compounds of methyl benzoate, benzene and toluene were used as the samples. And these samples,
A sample solution was prepared by dissolving it in methanol so as to have a concentration of 200 μg / mL, and 1 to 5 μL of this sample solution was injected into a liquid chromatography apparatus according to the detection sensitivity, and liquid chromatography was performed.

Then, the retention of each sample by each filler was investigated, and based on the retention, the separation coefficient of methyl benzoate or toluene with respect to benzene was calculated.

[0092]

[Table 3]

That is, first, in Table 3 above, the solute retention force given by each filler when various organic solvents are used as mobile phase solvents is shown by using the k'value as an index. Generally, when the retention characteristic of a column (filler) is exhibited, the time from the injection of a sample into the column to the top of the peak eluted from the column (retention time) is used. Since the relative evaluation can be performed only when the flow rate of the mobile phase solvent to be sent is the same and the columns used are the same, comparison of columns with different packing materials and column In the case where the sizes are different from each other, or when the liquid feeding rate of the mobile phase solvent is different, it becomes impossible to make a relative comparison even if the retention times are compared. Therefore, in the comparison of the results of the chromatographic separation under different conditions, the value represented by k ′ in the following formula is usually expressed, that is, the ratio of the retention time of the retained compound in the column to the elution time of the unretained solute. So, the retention characteristic (retention force) of each solute
Will be evaluated. k ′ = (t _r −t _o ) / t _o

Where t _{r is the} solute retention time, and t _o is the solute elution time that is not retained. Then, by using such a k ′ value, it is possible to ignore the influence of a difference in column length, thickness, flow rate, and the like, and thus it becomes possible to perform pure evaluation of the characteristics of the filler. is there.

The separation coefficient is a value representing the ratio of retention (k 'value) of two solutes, and is one of the scales indicating the degree of separation between peaks. Specifically, the separation factor = k'value of one solute / k'value of another solute. Therefore, when the separation coefficient or the reciprocal of the separation coefficient has a large value, it can be determined that the filler has a property of separating the two solutes well.

Further, as is clear from Table 3, the filler according to the present invention retains the compound having a hydrophobic group for a longer period because its hydrophobicity is strengthened. Specifically, the methyl group of toluene is a hydrophobic group, and the larger the separation coefficient of toluene for benzene that does not have a methyl group, the greater the ability of the packing material to recognize the hydrophobicity of the sample. The filler of the present invention example 2 (COP having a reduced reaction rate with respect to the carrier)
-5-2) except for the filler to which a stationary phase having a polycyclic aromatic functional group is bound (COP-5-1, PER-).
3, PYE, NE), the separation coefficient of toluene with respect to benzene tends to increase as the number of rings constituting the polycyclic aromatic functional group increases, which is a functional group chemically modifying silica gel. It was revealed that the more hydrophobic samples were retained longer as the number of rings in the polycyclic aromatic functional group increased.

Further, the filler according to the present invention is also enhanced in the retention of the compound having an electron-withdrawing group. Specifically, methyl benzoate has an electron-withdrawing functional group, and the larger the separation coefficient of methyl benzoate with respect to benzene, the greater the retention for a sample with an electron-withdrawing functional group. This shows that the filler (COP-5-1) of the present invention example 1 and the filler (PER) of the present invention example 3 (PER) were used more than when the filler (PYE) of the comparative example 1 was used.
When -3) is used, the separation coefficient of methyl benzoate with respect to benzene becomes larger, and the larger the number of rings constituting the polycyclic aromatic functional group bonded to silica gel as a carrier, the more electron withdrawing. It was revealed that the sample having a sexual functional group was retained longer.

As described above, the greater the number of rings constituting the polycyclic aromatic functional group contained in the stationary phase, the more the hydrophobic recognition ability and electronic interaction with the sample are improved. Thus, the compound can be efficiently separated by electronic interaction in addition to the separation ability by hydrophobicity. Therefore, by using the packing material according to the present invention, it is possible to effectively separate a compound group that is difficult to separate with a packing material that distinguishes and separates a sample mainly by its hydrophobicity such as a C ₁₈ column. It is presumed to obtain.

Example 3 In this example, a comparative examination will be made on the separation ability of various fillers for PCBs. Specifically, first, as a PCBs sample, 3,3 ′, 4,4′-tetrachlorobiphenyl, which is a PCB having no chlorine atoms at the 2- and 6-positions and substituted with 4 chlorine atoms, (IUP
AC No. 77) has one chlorine atom at the 2-position and 4
PCBs substituted with 4 chlorine atoms 2, 3, 4,
4'-tetrachlorobiphenyl (IUPAC No. 6
6) PCBs having no chlorine atoms at the 2- and 6-positions and substituted with 6 chlorine atoms: 3,3 ', 4,4', 5
5'-hexachlorobiphenyl (IUPAC No. 1
69) 2,2 ', 4,4', which is a PCB having 4 chlorine atoms at the 2 and 6 positions and substituted with 6 chlorine atoms.
6,6'-hexachlorobiphenyl (IUPAC N
o. 155) is prepared, and each of these has a concentration of 1
It was dissolved in toluene so as to have a concentration of mg / mL to prepare a sample solution, which was used by increasing or decreasing to about 1 μL to 10 μL according to the detection sensitivity of PCBs. The PCB sample used was provided by the Center for Disease Prevention and Prevention of the US Public Health Service, and the same applies to the examples below.

Further, as a packing material, the packing material shown in Table 4 below was packed in the same column as in Example 2 and liquid chromatography was carried out on the various sample solutions prepared above using the packing material. It was The apparatus used for liquid chromatography and the conditions therefor were the same as in Example 2. However, hexane was used as the mobile phase solvent, and the detection wavelength of the ultraviolet absorption detector was set to 220 nm.

As a result, the k'value, which is an index of the retention characteristic, was determined and shown in Table 4 below. Also, IUP
AC No. IUPAC for 66 PCB k'values
No. 77 PCB's k'value ratio, and IUPAC
No. IUPAC N for the PCB's k'value of 155
o. The ratio of the PCB's k'values of 169 was calculated as the separation factor and is also shown in Table 4 below.

[0102]

[Table 4]

As is clear from the results shown in Table 4, P that is highly toxic to PCBs that is not very toxic.
Separation coefficient of CBs, in other words, PCBs having chlorine atoms at the 2nd and 6th positions of the biphenyl skeleton, and PCBs having no chlorine atoms at the 2nd and 6th positions of the biphenyl skeleton
The separation coefficient of the class tends to increase as the number of rings of the polycyclic aromatic functional group contained in the stationary phase increases, and coronene or perylene is used as a carrier than PYE which has been conventionally used. Of the filler of the present invention (CO
P-5-1, PER-3) has a larger separation coefficient, and the filler according to the present invention retains highly toxic PCBs longer than non-toxic PCBs. , It was shown that the isomers of those PCBs can be effectively separated.

Further, in the filler in which coronene is bound to the carrier, PCBs can be advantageously separated even if the reaction rate between the stationary phase and the carrier is low. Specifically, for example, in the filler (COP-5-2) of Inventive Example 2, as shown in Table 1, the carbon content is 4%, but IUPAC N
o. 155 PCB for IUPAC No. 16
The separation factor of PCB of 9 is 20 or more, and the filler in which coronene is bound to the carrier has a low carbon content of about 4%, that is, even if the amount of the stationary phase bound to the carrier is small, Higher separation performance was obtained for PCB isomers than the type of packing.

Further, the filler (COP-5) of Example 1 of the present invention in which the polycyclic aromatic functional group (coronenyl group) was bound to the carrier by a spacer group composed of 5 methylene groups.
-1) was compared with the filler (PER-3) of Inventive Example 3 in which a polycyclic aromatic functional group (pyrenyl group) was bound to a carrier with a spacer group composed of three methylene groups. The spacer group that binds the polycyclic aromatic functional group and the carrier exhibits the ability to recognize PCB isomers, and the above-mentioned Example 2
Of the benzene derivative shown in Table 3 and the separation ability of the fullerene derivative shown in Example 6 which will be described later are less affected, and the fillers have toxicity that chlorine atoms are not bonded to the 2- and 6-positions. It is considered that the property of retaining strong PCBs for a longer period is expressed only by the property of the polycyclic aromatic functional group contained in the stationary phase. Therefore, the spacer group connecting the polycyclic aromatic functional group and the carrier may have a different chain length, or a functional group such as ester, amide or ether may be introduced into the spacer group in addition to the methylene group. Even if it is, it does not have a chlorine atom at the 2nd or 6th position,
It is presumed that highly toxic PCBs can be retained longer than non-toxic PCBs having chlorine atoms at the 2- and 6-positions.

Example 4 Separation of PCB isomers As described above, the toxicity of PCBs generally increases as the number of chlorine atoms bonded to the 2- and 6-positions of the biphenyl skeleton increases. However, in order to accurately evaluate the toxicity of PCBs contained in a sample, PCBs that do not have a chlorine atom at the 2nd and 6th positions and those that do not have a chlorine atom at the 2nd position are usually used. A filler that can separate PCBs having a chlorine atom at the 6-position is effective.

Therefore, in this example, the filler (COP-5-1) of Example 1 of the present invention retains highly toxic PCBs having no chlorine atom at the 2- and 6-positions for a longer time. , And it has chlorine atoms in the 2nd and 6th positions and is not highly toxic P
It is revealed that the separation from CBs can be effectively performed.

First, here, as the samples to be separated, PCBs having 4 to 6 chlorine atoms bonded are used, and they are classified into those having the same number of chlorine atoms bonded. Further, each of them has no chlorine atom bonded to the 2- and 6-positions of biphenyl (C group), one having one chlorine atom bonded to 2- or 6-position of biphenyl (M group), and biphenyl It was divided into three groups, that is, a group having two or more chlorine atoms bonded at the 2-position or 6-position (O group).

Next, these grouped PCBs
Each of the same types as in Example 3 had a concentration of 1 mg / mL.
A sample solution was prepared by dissolving in toluene so that the concentration became.

Then, as a packing material, the packing material of the present invention example 1 and the packing material of the comparative example 2 as a comparison object were packed in the same columns as in the above-mentioned embodiment example 2, and the packing material was used as described above. Liquid chromatography was performed on each of the prepared sample solutions, and the elution time of each is shown in Table 5 below. The apparatus used for liquid chromatography and the conditions thereof were the same as in Example 3, and the sample solution was used by increasing or decreasing to about 1 μL to 10 μL according to the detection sensitivity.

[0111]

[Table 5]

As is clear from the results shown in Table 5, when the filler (COP-5-1) of Example 1 of the present invention was used, PCB isomers having the same number of bonded chlorine atoms were bound to each other. Comparing with, the elution time of the PCB isomer (group C) having no chlorine atoms at the 2nd and 6th positions is 2
It was shown that the elution time of PCB isomers having a chlorine atom at the 6-position or 6-position (group M, group O) was longer, and the groups C, M and O could be advantageously separated. . On the other hand, the filler of Comparative Example 1 (P
When YE) was used, PCB isomers having the same number of bonded chlorine atoms were compared with each other and the elution times of PCB isomers having no chlorine atoms at the 2nd and 6th positions (group C) were compared. The elution time of PCB isomers having a chlorine atom at the 2- or 6-position (group M, group O) showed no significant difference, indicating that the separation was insufficient.

FIGS. 1 and 2 are chromatograms obtained by simultaneously separating an isomer mixture of 72 PCBs using the packing material of Example 1 of the present invention and the packing material of Comparative Example 1, respectively. showed that. In addition, in FIG. 1 and FIG. 2, the numbers above the peaks indicate the IUPAC No. of the PCBs giving the peaks. Is represented. The liquid chromatography apparatus and conditions are the same as above, and the sample solution is IUPAC No. But 2,3,11,12,1
3,14,15,22,28,33,35,36,37,38,39,41,44,47,49,52,56,
60,66,74,77,78,79,80,81,87,99,101,105,110,114,118,
122,126,127,130,137,138,146,151,153,154,156,157,15
8,167,169,170,172,174,177,178,180,182,183,187,189,
191,193,194,195,196,197,201,203,204,206,209 PCB standard product with toluene, concentration of 500 μL / m
Diluted to L and injected a total of 8 μL.

As is apparent from FIGS. 1 and 2, the use of the filler (COP-5-1) of Example 1 of the present invention resulted in a highly toxic IUPAC No. 77,
The PCBs of 126 and 169 are completely separated, while the filler of Comparative Example 1 (PYE) is used.
Poor retention of the highly toxic PCBs results in incomplete separation of them.

Further, in Table 5 above, when liquid chromatography was carried out using the filler (COP-5-1) of Inventive Example 1, the number of bonded chlorine atoms was 4
Among the individual PCBs, the elution time was partially overlapped between the highly toxic group C and the less toxic group M or group O, indicating that the groups were not completely separated. However, as shown in FIG. 1, when liquid chromatography was performed using the packing material of Example 1 of the present invention, among the PCB isomers having 4 bonded chlorine atoms, And the most toxic 3,3 ',
4,4'-Tetrachlorobiphenyl (IUPAC N
o. 77) was almost completely separated, indicating that highly toxic PCB isomers can be effectively separated.

Example 5 In this example, in order to demonstrate that it is possible to effectively separate fullerenes by using the filler according to the present invention, according to the existing filler and the present invention. The holding power of fullerene and the separation coefficient when a filler was used were compared, and the results are shown in Table 6 below.

In this example, a fullerene mixed powder (trade name: Refined C) was used as a sample used for evaluating the retention property of the fullerene by the filler.
Fullerene obtained by separating and refining ( ₆₀ / C ₇₀ : manufactured by vacuum metallurgy) was used. Specifically, the fullerene mixed powder is obtained by extracting a fullerene allotrope or isomer from a soot containing fullerenes and then drying.
In addition to C ₆₀ and C ₇₀ , a small amount of high-order fullerenes are contained.
₆₀ and C ₇₀ were separated and purified to obtain standard products. Then, the obtained standard fullerene was diluted with toluene so as to have a concentration of about 0.3 to 1 mg / mL to prepare a sample solution. Also,
The analysis was performed by injecting 1 μL to 5 μL of the prepared standard sample as needed using a microsyringe. Toluene was used as the mobile phase solvent. However, since the C ₁₈ type filler cannot retain fullerenes when a mobile phase solvent of toluene is used, a C ₁₈ type filler is a poor solvent for fullerenes only when the C ₁₈ type filler is used as the filler. Some hexane was used. The fullerene detection wavelength is 285n.
m.

[0118]

[Table 6]

As is clear from the results shown in Table 6, fullerene can be effectively separated by using the filler according to the present invention. That is, in the fillers of Comparative Examples 1 to 3 which have been conventionally used, the separation coefficient of C ₇₀ with respect to C ₆₀ is 2 or less, whereas in the case of using the fillers of Examples 1 to 3 of the present invention, It was confirmed that each of them has a separation coefficient of 2 or more and has a characteristic of discriminating a difference in the number of constituent carbon atoms of fullerenes higher than that of the fillers used conventionally.

[0120]

As is apparent from the above description, in the filler according to the present invention, not only hydrophobic interaction but also
Since it has an interaction with an electron-withdrawing group and can exert a strong retention force for a solute, it is a C ₁₈ -type packing material having a hydrophobic interaction as a main separation ability under reverse phase chromatography conditions. It can be advantageously applied to substances that cannot be retained or separated, for example, substances such as PCBs and fullerenes, and can effectively retain and separate them in a liquid chromatography operation.

According to the method for producing a packing material for liquid chromatography according to the present invention, the excellent liquid chromatography as described above is carried out by reacting a silylating agent having a polycyclic aromatic functional group with a hydroxyl group on the carrier surface. It is possible to easily manufacture by making it.

[Brief description of drawings]

FIG. 1 is a chromatogram showing the result of separating an isomer mixture of PCBs by liquid chromatography using the packing material according to the present invention.

FIG. 2 is a chromatogram showing the result of separating an isomer mixture of PCBs by liquid chromatography using the packing material of Comparative Example.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display location G01N 30/88 G01N 30/88 C

Claims (5)

[Claims] 1. A packing material for liquid chromatography comprising a carrier chemically modified with a polycyclic aromatic functional group having 5 to 10 condensed rings. 2. The carrier is silica gel.
The packing material for liquid chromatography according to item 1. 3. A silylating agent having a polycyclic aromatic functional group having 5 to 10 condensed rings is used, and the silylating agent is reacted with a hydroxyl group on the surface of a carrier to give the polycyclic aromatic functional group. A method for producing a packing material for liquid chromatography, which comprises forming a stationary phase having a group on the surface of the carrier. 4. The carrier is silica gel.
The method for producing a packing material for liquid chromatography as described in 1. 5. A packing composed of a carrier chemically modified with a polycyclic aromatic functional group having 5 to 10 condensed rings when separating polychlorinated biphenyls or fullerenes by a liquid chromatography method. A method for separating, which comprises using an agent.