A kind of reversed-phase liquid chromatography detects the method for raltitrexed enantiomer
A technology of reversed-phase liquid chromatography and enantiomers, which can be used in measuring devices, instruments, scientific instruments, etc. Volatility and other issues
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Embodiment 1
[0031] The detection of enantiomer in embodiment 1 raltitrexed
[0032] 1. Instruments and reagents
[0033] Shimadzu LC-20A high performance liquid chromatography;
[0034] CPA225D electronic balance (Beijing Sartorius Instrument System Co., Ltd.);
[0035] Chromatographic column: Waters XBridge C18 column;
[0036] The raw material of raltitrexed was purchased from Wuhan Dahua Weiye Pharmaceutical Chemical Co., Ltd., and the reference substance of raltitrexed and enantiomer was purchased from EP;
[0037] Methanol was chromatographically pure, triethanolamine was analytically pure, water was deionized water, and the mobile phase was prepared and filtered through a 0.45 μm membrane.
[0038] 2. Experimental methods and results
[0039] 1. Solution preparation
[0040] Diluent: 42% methanol aqueous solution, add 5mL triethanolamine to 1L methanol aqueous solution.
[0041] Reference substance solution: Accurately weigh approximately 10 mg of raltitrexed and its enantiome...
Embodiment 2
[0065] Example 2 failure case
[0066]The applicant has tried a variety of mobile phases of common composition, such as methanol aqueous solution and acetonitrile aqueous solution of different concentrations, but no obvious effect has been seen. Taking the 42% methanol aqueous solution (compared with Example 1 without adding triethanolamine, and other chromatographic parameters are consistent) as an example, the chromatographic peaks of raltitrexed and enantiomers are always co-eluted together, cannot be separated. Usually, when encountering this situation, technicians will consider whether the proportion of methanol is too high, and try to separate the two by reducing the proportion of methanol. After many attempts by the applicant, when using 36% methanol aqueous solution for elution, although there is a certain sign of separation between the two, the peak spread is very serious, the retention time is also significantly prolonged, and the number of theoretical plates and ta...
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