JPH0565501B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel oxalic acid bisester derivatives useful as luminescent reagents for fluorescent substances. Currently, bis(2,
Oxalic acid bisesters such as 4,6-trichlorophenyl) oxalate and bis(6-carbopentoxy, 2,4,5-trichlorophenyl) oxalate (hereinafter abbreviated as CPPO) are commonly used. There is. For long-term luminescence, CPPO, which has high solubility and large luminescence ability, is often used. However, even with CPPO, crystals may precipitate when the winter storage temperature drops, resulting in weaker luminescence. In order to maintain strong luminescence for a long time even when used in cold regions, it became necessary to develop oxalic acid bisesters that have higher solubility and greater luminescent ability. In the case of chemical lights, phthalate esters are the most suitable solvents for oxalate bisesters, so the inventors investigated various oxalate bisester derivatives that have high solubility in phthalate esters, and have developed the present invention. Achieved the invented compound and completed the present invention. That is, the present invention is based on the general formula _{[ In the formula ,} It is an invention. n may be any number from 1 to 12, but preferably n = 2 to 6; m may be any number from 1 to 5, but preferably m = 1 to 3. The novel oxalic acid bisester derivative of the present invention has a higher solubility in phthalate esters such as dibutyl phthalate (hereinafter abbreviated as DBP), which is suitable for reaction with hydrogen peroxide in chemical light, than the above-mentioned CPPO. When using this as the luminescent reagent of the present invention, the concentration of the active intermediate can be higher than that of CPPO, and as a result, a luminescent reaction with high brightness can be obtained for a long time. Therefore, the present invention provides a new and more effective luminescent reagent that maintains high brightness for a longer period of time than conventional products in the field of chemical lights and the like that utilize this luminescent reaction. Table 1 shows the solubility of two examples of the compounds of the present invention in various solvents at 20°C in comparison with that of CPPO. The numbers in the table indicate the number of grams of sample that can be dissolved in 100 ml of solvent. (Hereafter, compound (1) in the table is
EMPO and (2) are abbreviated as BMPO. ) Table 1 is shown in the attached sheet. Furthermore, Table 2 shows the solubility in DBP at 0°C.

【table】

[Table] Describing the synthesis method of the present invention, 2, 4, 5
- Using trichlorosalicylic acid as a starting material, it is converted into C _o H _2o+1 - (OC ₂ H ₄ ) _n using an esterification catalyst such as para-toluenesulfonic acid, thionyl chloride, aluminum chloride, or boron trifluoride ether complex. React with monoalkyl ether of glycols called -OH,
6-alkylethylene (or alkyl oligoethylene) glycoxycarbonyl-2,4,5
-Trichlorophenol can be easily synthesized by a two-step reaction in which trichlorophenol is obtained and reacted with oxalyl chloride using triethylamine as a catalyst. The synthesis method will be explained in detail below using Examples. Example 1 (1) 17 g of 2,4,5-trichlorosalicylic acid
(0.07 mol), dry ethylene glycol monoethyl ether 126 g (1.4 mol), and para-toluene sulfonic acid 2.4 g (0.014 mol) were mixed and refluxed at 135°C, while removing the water produced in a Dean-stark water separator. The reaction will complete in 10 hours, so after cooling, add 400 ml of water and extract with ether. The ether solution was washed with sodium bicarbonate water, dried, concentrated under reduced pressure to remove the solvent, and further fractionated under reduced pressure to obtain 15 g of a milky white solid of 6-(ethyl monoglycoxycarbonyl)-2,4,5-trichlorophenol. was gotten. Yield 83.3%, bp145℃/0.4mmHg, m.
p.58~60℃, IR (KBr) 1750cm ^-1
(

[Formula]), 1120cm ^-1 (

[Formula]), 3250cm ^-1 (φ-OH) (2) 6-(ethylmonoglykoxycarbonyl)- obtained as a solid by the reaction in the first step above
2,4,5-trichlorophenol 6.2g
(0.02 mol) was dissolved in 60 _ml of dry toluene, 2 g (0.02 mol) of triethylamine was added, and under stirring and cooling, oxalyl chloride 3
g (0.024 mol) was added dropwise at 5 to 10°C, and then reacted at room temperature for 3 hours. After the reaction, the precipitated triethylamine hydrochloride was filtered off and concentrated under reduced pressure to obtain light brown crystals of the target bis{6-(ethylmonoglycoxycarbonyl)-2,4,5-trichlorophenyl}oxalate 6.0 I got g. Yield 89%, mp113~115℃, IR (KBr) 1810cm ^-1 (

[Formula]), 1760cm ^-1 (

[Formula]) NMR (CDCl ₃ ) δ: 1.2 (6H, -CH ₂ C H ₃ ), 3.6 (8H, -O-C H ₂
−), 4.5 (4H,

[Formula]), 7.7 (2H, φ- H ) UV absorption (0.02mM acetonitrile solution): λmax 212mμ, ε5.5×10 ⁴ Luminescence test positive Note Luminescence test method Solution A: Bis(phenylethynyl)anthracene 1
mg dissolved in 100 ml of acetone Solution B: H ₂ O ₂ water 0.01 molar acetone solution Solution C: 1 ml each of PH4 buffers A, B, and C were mixed, and when the sample was poured into this, it emitted green light in the dark. . Example 2 (1) 17 g of 2,4,5-trichlorosalicylic acid
(0.07 mol), 166 g (1.4 mol) of dry ethylene glycol monobutyl ether, and 12 g (0.085 mol) of boron trifluoride ether complex.
The reaction was carried out at 70°C for 20 hours, left to cool, concentrated under reduced pressure to remove the solvent, and further fractionated under reduced pressure to obtain 6-(butylmonoglycoxycarbonyl)-2,4,5-
14 g of a pale yellow oil of trichlorophenol was obtained. Yield 69.0% IR (neat) 1760cm ^-1 (

[Formula]), 1120cm ^-1 (

[Formula]), 3400cm ^-1 (φ-OH), bp 150℃/0.15mmHg (2) 6-(Butylmonoglykoxycarbonyl)-2,4,5-trichlorophenol obtained above 6.8
g (0.02 mol), reacted and treated in the same manner as in (2) of Example 1 to obtain bis{6-(butylmonoglycoxycarbonyl)-2,4,5-trichlorophenyl}oxalate (hereinafter referred to as , abbreviated as BMPO) was obtained. Yield 95.0% mp66−68℃, IR (KBr) 1805cm ^-1 (

[Formula]), NMR (CDCl ₃ ) δ: 0.9 {6H, −(CH ₂ ) ₃ −CH ₃ }, 1.4 (8H, −CH ₂ −
CH2 _- CH2 _{- CH3} ), 3.5(8H, _-O - CH2
−), 4.5 (4H,

[Formula]), 7.7 (2H, φ − H ), U absorption (0.02mM acetonitrile solution): λmax 212mμ, ε5.4×10 ⁴ Luminescence test positive example 3 (1) 2,4,5-trichlorosalicylic acid 17g
(0.07mol), dry benzene 100ml, pyridine 5
The drops were mixed and 12 g (0.1 mol) of thionyl chloride was added dropwise to this at 55-60° C. while stirring. After dripping
After reacting at 60-65°C for 2 hours, concentrate under reduced pressure to remove benzene and excess thionyl chloride.
Immediately add 250 g (1.4 mol) of triethylene glycol monoethyl ether, cool to 20°C, and add 7.0 g of triethylamine while stirring.
(0.07 mol) was added dropwise. 40~50℃ after completion of dripping
After the reaction was completed, the triethylamine hydrochloride produced was filtered and the solution was distilled under reduced pressure to produce 6-(ethyl triglycoxycarbonyl).
26 g of yellow oil of -2,4,5-trichlorophenol was obtained. Yield 89%, bp200℃/0.35mmHg, IR (neat)1755cm ^-1 (

[Formula]), 1120cm ^-1 (

[Formula]), 3450cm ^-1 (φ−OH). (2) 6-(ethyltriglykoxycarbonyl)-2,4,5-trichlorophenol obtained above 8.0
g (0.02 mol) and treated in the same manner as in Example 1 (2) to obtain white crystals of bis{6-(ethyltriglycoxycarbonyl)-2,4,5-trichlorophenyl}oxalate. I got it. Yield 93.3%, IR (KBr) 1805cm ^-1 (

[Formula]), 1740cm ^-1 (

[Formula]), NMR (CDCl ₃ ) δ: 1.2 (6H, -CH ₂ -C H ₃ ), 3.6 (24H, -O-C H ₂
−), 4.5 (4H,

[Formula]), 7.7 (2H, φ- H ) UV absorption (0.02mM acetonitrile solution): λmax 212mμ, ε5.3×10 ⁴ Luminescence test positive. Next, BMPO, which is one of the luminescent reagents of the present invention,
The luminous performance will be shown in Examples. Example 4 Adding hydrogen peroxide to 3-methyl-3-pentanol
Take 50μ of a solution containing 5M and 1mM sodium salicylate, add 200μ of a 15mM dibutyl phthalate solution of 9,10-bis(phenylethyl)anthracene and BMPO (or CPPO).
When 1 ml of a 100 mM dibutyl phthalate solution was added in this order, a green luminescent reaction started, and the luminescent intensity was immediately measured at fixed time intervals using a photometer. The measurement temperature was 20°C, and the results are shown in Figure 1. Example 5 Adding hydrogen peroxide to 3-methyl-3-pentanol
Take 50μ of a solution containing 5M and 1mM sodium salicylate, and add 1-chloro-
9,10-bis(phenylethyl)anthracene
Add 200μ of 35mM dibutyl phthalate solution and
Keep at ℃. Next, when 1ml of a 210mM dibutyl phthalate solution of BMPO (or CPPO) (stored at 0℃ for 1 month after making the solution) is added to the previous two mixtures, a yellow luminescence reaction begins, and the luminescence can be immediately detected using a photometer. The strength was measured. However, a small amount of precipitate was observed in CPPO. The measurement temperature was 0° C., and the obtained results are shown in FIG. As is clear from Figure 1, BMPO has the same luminescence intensity as CPPO in the initial stage, but the high-intensity luminescence lasts longer than CPPO, until about 4 hours later.
Emission intensity is higher than that of CPPO. Furthermore, as is clear from FIG. 2, at 0° C., unlike CPPO, BMPO does not precipitate during storage, so there is a clear difference in luminescence intensity. In this way, BMPO has excellent performance as a luminescent reagent used in chemical lights. In addition, the luminescent reagent (BMPO, etc.) of the present invention dissolves very well in the solvent used for liquid chromatography (acetonitrile, acetone, etc.) and has high luminescence sensitivity, so it can also be used as a luminescent reagent for liquid chromatography. It is applicable.

[Brief explanation of the drawing]

FIG. 1 is a graph showing the change in luminescence intensity over time at 20° C. for BMPO, which is a compound of the present invention, and existing CPPO in Example 4. (However, 〓〓 is about BMPO, and 〓〓
〓 indicates the measurement results for CPPO. ) The vertical axis shows relative luminescence intensity, and the horizontal axis shows time. Figure 2 shows the results for BMPO, the compound of the present invention, and existing CPPO in Example 5.
The change in luminescence intensity with time at 0% after storage at 0.degree. C. is shown in the same manner as in FIG. 1.

Claims (1)

[Claims] 1. General formula [In the formula, X is C _o H _2o+1 −(OC ₂ H ₄ ) _n − (however, n represents 1 to 12 and m represents 1 to 5). ]