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Efficient liquid chromatogram detection method of L-leucine

A technology of high performance liquid chromatography and detection method, which is applied in the field of high performance liquid phase detection of L-leucine, which can solve the problems of long detection time and high requirements for reagents, achieve fast peak time, shorten detection time, and save costs Effect

Active Publication Date: 2018-09-14
JING JING PHARMA
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Problems solved by technology

The prior art discloses the use of C 18 Chromatographic column, 0.01mmol / L potassium dihydrogen phosphate: mixed mobile phase of acetonitrile and methanol, online derivatization method, but the detection time of this method is long, a sample needs 35min, and gradient elution is required. Reagents for preparing mobile phases have higher requirements

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  • Efficient liquid chromatogram detection method of L-leucine
  • Efficient liquid chromatogram detection method of L-leucine
  • Efficient liquid chromatogram detection method of L-leucine

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Embodiment 1-3

[0045] 1. Chromatographic conditions:

[0046] Chromatographic column: amino bonded silica gel column;

[0047] Mobile phase: the volume ratio of potassium dihydrogen phosphate aqueous solution to acetonitrile is 40:60; (among them, the preparation of potassium dihydrogen phosphate aqueous solution is: take 13.61g of potassium dihydrogen phosphate, add 500ml of water to dissolve, add 5ml of 25-28% ammonia water , add water to dilute to 1000ml, mix well and then adjust the pH value to 4.2±0.02 with phosphoric acid);

[0048] Mobile phase flow rate: 0.7mL / min;

[0049] Detection wavelength: 210nm;

[0050] Chromatographic column temperature: 30°C.

[0051] 2. Experimental steps

[0052] (1) prepare mobile phase, filter, degas;

[0053] (2) adjust the high performance liquid chromatograph, and balance the chromatographic column;

[0054] (3) preparation of reference substance solution and need testing solution, take by weighing 20.0mg of L-leucine reference substance in 50m...

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Abstract

The invention discloses an efficient liquid chromatogram detection method of L-leucine, and relates to the technical field of column chromatographic methods. Chromatographic conditions are that the chromatographic column is an amino-bonded silica gel column, the mobile phase is a monopotassium phosphate aqueous solution and acetonitrile with the volume percentage being 37%:63%-42%:58%, the concentration of the monopotassium phosphate aqueous solution is 0.1-0.2 mol / L, ammonia water is contained in the monopotassium phosphate aqueous solution and accounts for 0.5 vt% of the monopotassium phosphate aqueous solution, and the pH value of the monopotassium phosphate aqueous solution is 4.2+ / -0.02; the flow rate of the mobile phase is 0.7-1.0 mL / min, the detection wavelength is 210 nm, and the column temperature of a chromatographic column is 30 DEG C; efficient liquid chromatogram analysis is conducted, and the L-leucine content in conversion fluid is measured. Accordingly, the L-leucine content can be measured rapidly and accurately, it is ensured that the quality is controllable, the error is reduced, the cost is reduced, operation is easy, the detection time is short, the accuracy ishigh, and the efficiency is improved.

Description

technical field [0001] The invention relates to the technical field of column chromatography, in particular to a high-performance liquid phase detection method for L-leucine. Background technique [0002] Calcium Ketoleucine works in the body by converting the nitrogen of non-essential amino acids into leucine. Leucine is one of the eight essential amino acids for the human body and belongs to branched-chain amino acids. L-leucine is an indispensable raw material for compound amino acid intravenous injection, which plays an active role in maintaining the nutritional needs of critically ill patients and saving their lives. At the same time, as a branched-chain amino acid, because of its fast oxidation rate, the efficiency of decomposing and producing ATP in the body is higher than other amino acids, so it can be used to supplement the energy consumption caused by large amounts of exercise, reduce the fatigue response of the central nervous system caused by it, and reduce hyp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 王阳何连顺李斌水米造吉马静
Owner JING JING PHARMA
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