Preparation and its use of derivatization reagent for detecting L-carnitine or D-carnitine
OwnerShuhua Gu, Qingyi Li +1inventors
InventorsShuhua Gu, Qingyi Li, Xuecheng Wang
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Abstract
A preparation method and its use of derivatization reagent for detecting L-carnitine or D-carnitine are provided. The present reagent is stable. It can be used for detecting L-carnitine or D-carnitine accurately and sensitively. That is to say, the reagent is applied to detecting the amount of synthesized or natural L-carnitine and the amount of mixing D-carnitine. The compound reagent is used for determining the chiral isomers of chemicals, biological reagents, health care reagents, cosmetic, body fluids and various foods, which contain L-carnitine or/and D-carnitine, and optical isomers of other chiral amino acids.
Claims14
A method for detecting the content of L-carnitine or D-carnitine in a sample comprising L-carnitine or D-carnitine, wherein the method includes following steps:
- (a) preparing a solution comprising the sample containing a proper amount of L-carnitine (or D-carnitine) and a control solution containing DL-carnitine;
- (b) mixing a proper amount of a derivative reagent and the test sample solution containing L-carnitine (or D-carnitine), and allowing them to react to generate L-carnitine (or D-carnitine) derivatives;
- wherein the derivative reagent has a structure of:
- wherein:
- the carbon atom marked with an asterisk is the chiral carbon atom;
- the compound is chiral compound having pure optical active, the D- or L-compound;
- R represents C1-C6 straight-chain or branched alkyl groups, C6-C10 aryl groups, C2-C6 straight-chain or branched alkenyl or alkynyl groups or C3-C6 cycloalkyl groups;
- and
- X represents a halogen atom;
- (c) subjecting the reacted test sample solution to HPLC to detect and thereafter calculating the content of L-carnitine (or D-carnitine) in the sample.
The method according to claim 1, wherein the method further includes following steps:
- (d) preparing the derivatization reagent solution:
- a D- or L-optical pure compound of formula (I) of claim 1 is dissolved in solvent of claim 6 to form a 0.01˜100 mg/ml solution under the dark conditions, wherein the compound of formula (I) is (+)α-methyl-6-methoxy-2-naphthyl acetyl chloride, and the solvent is acetonitrile;
- the concentration of solution is from 1 to 10 mg/ml;
- (e) preparing a test solution of L-carnitine or D-carnitine, and a control solution of DL-carnitine;
- (f) causing the derivatized reagent of step (d) to mix and react with the test solution and control solution of step (e) respectively in a airtight vessel, in the present of a solvent at 20° C.˜95° C. in a water bath for 20-180 min;
- and
- (g) using high performance liquid chromatography (HPLC) to separate and detect the reacted test solution and control solution, thereafter the content of L-carnitine (or D-carnitine) in test solution is calculated using an external standard method.
Dependencies are inferred from the claim text.