[논문]A Convenient HPLC/ELSD Method for the Quantitative Analysis of Betaine in Lycium chinense

Lee, Sang-Myung; Park, Chae-Kyu; Cho, Byung-Goo; Cho, Kyoung-Shim; Min, Byung-Sun; Bae, Ki-Hwan

A Convenient HPLC/ELSD Method for the Quantitative Analysis of Betaine in Lycium chinense 원문보기

Natural product sciences, v.17 no.2, 2011년, pp.104 - 107

Lee, Sang-Myung (Korea Ginseng Corp. Central Research Institute) , Park, Chae-Kyu (Korea Ginseng Corp. Central Research Institute) , Cho, Byung-Goo (Korea Ginseng Corp. Central Research Institute) , Cho, Kyoung-Shim (Korea Ginseng Corp. Central Research Institute) , Min, Byung-Sun (College of Pharmacy, Catholic University of Daegu) , Bae, Ki-Hwan (College of Pharmacy, Chungnam National University)

Abstract ▼ AI-Helper

In order to facilitate the quality control of betaine from the fruits of Lycium chinense, we have developed a rapid and simple method for quantitative determination. Determination was achieved on a Discovery C18 column with an isocratic solvent system of 0.32% perfluoropentanoic acid aqueous-acetonitrile at a flow-rate of 0.5 mL/min and detected an ELSD. The method was reproducible with intra- and inter-day variations of less than 6% (R.S.D). The recoveries were in the range of 90.01~100.05%. The method turned out to be fast and simple, furthermore, to have a good selectivity and sensitivity for the quantity determination of betaine in the fruits of L.chinense.

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Each standard was analyzed three times. This repeatability was chosen in order to determine the accuracy of the method. For this, three concentrations (0.

제안 방법

The recovery test for improving the accuracy of this method will be described with the next procedure. After adding the standards which correspond to 60, 100 and 140% of the amounts of betaine in the L. chinense fruit powder, the sample solutions were prepared according to the method in section - Preparation of sample and standard solutions. The quantity of each analyte was subsequently obtained from the corresponding calibration curve.
This repeatability was chosen in order to determine the accuracy of the method. For this, three concentrations (0.25, 0.5, 1 g/100 mL) and three replicates of each concentration were sampled for the repeatability test. The limits of detection (LOD) and quantification (LOQ) for betaine was determined at the standard deviation of the y-intercepts of the regression lines to the slope of the calibration curves ratio (s/S) of 3.
Identification by MS/MS detector − The Waters ACQUITY UPLC with a TQ Detector system was used to confirm the method specificity.
Method Validation − The identification of betaine was carried out by comparing the retention times of the peak in the standard with the corresponding ones in the sample solutions measured through the HPLC-ELSD method, and the quantitative values of betaine measured through the HPLC-ELSD method and the UPLC-MS/MS method.

대상 데이터

The standard material of betaine (> 95%) was purchased from ChromaDex (CA. USA), perfluoropentanoic acid (97%) from Sigma-Aldrich (WI, USA) and acetonitrile (HPLC grade) was obtained from J. T. Baker (NJ, USA).
chinense were a contracted cultivation by Korea Ginseng Corporation (Daejon, Korea). The voucher specimens were deposited in the specimen custody freeze warehouse of KGC Central Research Institute (Daejon, Korea). The standard material of betaine (> 95%) was purchased from ChromaDex (CA.

성능/효과

Therefore, this method showed great reliability on obtaining accurate results. All the results above indicated that this analysis method was rapid, precise, accurate, and sensitive for quantitative determination of betaine from fruits of L. chinense.
From the results, the R.S.Ds of the overall recoveries were < 5% and the recovery rates were at 90.01~100.05% which showed that the method was high accuracy.
2). In particular, the quantitative values by the ELSD and the MS/MS detector were almost identical with an inaccuarcy of 3.79 %. Thus, the analysis of the specificity of betaine was confirmed.

참고문헌 (13)

Barak, A.J., Harriet, C., Beckenhauer, H.C., Matti, J., and Dean, J.T., Dietary Betaine Promotes Generation of Hepatic SAdenosylmethionine and Protects the Liver from Ethanol-Induced Fatty Infiltration. Alcohol. Clin. Experiment. Res. 17, 552-555 (1993).
Funayama, S., Yoshida, K., Konno, C., and Hikino, H., Structure of kukoamine A, a hypotensive principle of Lycium chinense root barks1. Tetrahedron Lett. 21, 1355-1356 (1980).

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Gahl, W.A., Bernardini, I., and Chen, S., The effect of oral betaine on vertebral body bone density in pyridoxine-nonresponsive homocystinuria. J. Inherit. Metabol. Dis. 11, 291-298 (1988).
Junnila, M., Barak, A.J., Beakenhauer, H.C., and Rahko, T., Betaine reduces hepatic lipidosis induced by carbon tetrachloride in Sprague- Dawley rats. Veter. Human Toxicol. 40, 263-266 (1998).
Kikuchi, N., Matsuno, K., and Miki, K., Separation and determination of betaine in an oriental medicine by liquid chromatography. Analytica Chimica Acta 283, 338-343 (1993).

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Murakami, T., Nagamura, Y., and Hirano, K., The Recovering Effect of Betaine on Carbon Tetrachloride-Induced Liver Injury. J. Nutrit. Sci. Vitaminol. 44, 249-255 (1998).
Mar, M.H., Ridky, T.W., Garner, S.C., and Ziesel, S.H., A method for the determination of betaine in tissues using high performance liquid chromatography. J. Nutrit. Biochem. 6, 392-398 (1995).
Noguchi, M., Mochida, K., Shingu, T., Kozuka, M., and Fujitani, K., Uber die Bestandteile der chinesischen Droge "Ti-ku-'pi."I. Isolierung und Konstitution von Lyciumamid, einem neuen Dipeptid. Chem. Pharm. Bull. 32, 3584-3587 (1984).

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Selhub, J., Homocysteine metabolism. Ann. Rev. Nutrit. 19, 217-246 (1999).
Shin, Y.G., Cho, K.H., Kim, J.M., Park, M.K., and Park, J.H., Determination of betaine in Lycium chinense fruits by liquid chromatography-electrospray ionization mass spectrometry. J. Chromatograph. A 857, 331-335 (1999).
Sannai, A., Fujimori, T., and Kato, K., Neutral Volatile Components of "Kukoshi" (Lycium chinense M.). Agricul. Biol. Chem. 47, 2397-2399, (1983).
Wendel, U. and Bremer, H.J., Betaine in the treatment of homocystinuria due to 5,10-methylenetetrahydrofolate reductase deficiency. Eur. J. Pediatrics 142, 147-150 (1984).
Yang, L.L., Yen, K.Y., Kiso, Y., and Kikino, H., Antihepatotoxic actions of formosan plant drugs. J. Ethnopharmacol. 19, 103-110 (1987).

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A Convenient HPLC/ELSD Method for the Quantitative Analysis of Betaine in Lycium chinense 원문보기

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A Convenient HPLC/ELSD Method for the Quantitative Analysis of Betaine in Lycium chinense 원문보기

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