Abstract��
OBJECTIVE To develop an HPLC-MS/MS method coupled with EZ:faastTM amino acid analysis kits for the determination of glutamine in rat plasma for the pharmacokinetic study in rats.METHODS The plasma samples were prepared by EZ:faastTM amino acid analysis kits and then analyzed with API 3000 HPLC-MS/MS system. The analytical column was Phenomenex EZ:faast 4 �� AAA-MS(4 micron, 3.00 mm��250 mm) and the column temperature was 20 ��. The mobile phase was composed of 0.2% formic acid aqueous solution(containing 5 mmol��L-1 ammonium acetate)-methanol and eluted gradiently at a flow rate of 0.4 mL��min-1. The detection was performed with multiple reactions monitoring(MRM) using positive electrospray ionization(ESI). RESULTS The calibration curve for glutamine was linear over the concentration range of 3.003-150.2 ��g��mL-1(r=0.996 6). The lower limit of quantification was 3.003 ��g��mL-1. The inter- and intra-day RSDs were less than 15% and the accuracy was within 85%-115%. The extraction recoveries were around 90%, and glutamine was proved to be stable under different circumstances.CONCLUSION This method is specific, sensitive, and accurate, and the sample preparation procedure is simple, rapid, and suitable for the pharmacokinetic study of glutamine in rats.
WANG Ting,
JIANG Xue-Hua,
WANG Ling-*
.HPLC-MS/MS Method for Determination of Glutamine in Rat Plasma and Application to Pharmacokinetic Study[J] Chinese Pharmaceutical Journal, 2015,V50(3): 253-257
��
[1]
GENG J, BI J X, ZENG A P, et al. Application of hybrid cybernetic model in simulating myeloma cell culture co-consuming glucose and glutamine with mixed consumption patterns. Proc Biochem, 2013, 48(5): 955-964. [2] MANSOUR A, MOHAJERI-TEHRANI M R, QORBANI M, et al. Effect of glutamine supplementation on cardiovascular risk factors in patients with type 2 diabetes. Nutrition, 2015,31(1):119-126.[3] LI N, LEWIS P, SAMUELSON D, et al. Glutamine regulates Caco-2 cell tight junction proteins. Am J Physiol Gastrointest Liver Physiol, 2004, 287(3): 726-733.[4] NEWSHOLME P. Why is L-glutamine metabolism important to cells of the immune system in health, postinjury, surgery or infection. J Nutrition, 2001, 131(9): 2515-2522.[5] ASRANI V, CHANG W K, DONG Z, et al. Glutamine supplementation in acute pancreatitis: A meta-analysis of randomized controlled trials. Pancreatology, 2013, 13(5): 468-474.[6] MADL J E, DUNCAN C G, STANHILL J E, et al. Oxidative stress and redistribution of glutamine synthetase in california sea lionswith domoic acid toxicosis. Jcomppathol, 2014, 150(2): 306-315.[7] WERNERMAN J. Role of glutamine supplementation in critically ill patients. Curr Opin Anesthesiol, 2008, 21(2): 155-159.[8] ARCHIBALD R M. The enzymatic determination of glutamine. J Biol Chem, 1944, 154(3): 643-656.[9] MOHYI EL-DIN A R, DAVID M G.Anenzymicmicro-method for determination of glutamine and asparagine in blood. Anal Bio Chem,1963, 6(2): 144-152. ZIEGLER T R, BENFELL K, SMITH R J, et al. Safety and metabolic effects of L-glutamine administration in humans. J Parenter Enteral Nutr, 1990, 14(4 suppl): 137-146. TANG J L, ZHOU S W, YING Y,et al. Experimental evaluation of the bioequivalence of compound glutamine granules. Chin Pharmacol Bull(�й�ҩ��ѧͨ��), 2005, 21(12): 1509-1513. JIA S S, LI Z N, ZHAO Y,et al.HPLC Determination and application of glutamine content in rats intestines mucous membrane tissues.Lab Med Clin(����ҽѧ���ٴ�), 2011,8(1): 16-17. ZHU M W, XIAO L Y, LIU L,et al.The pharmacokinetics study of glutamine granules in healthy volunteers.Chin J Clin Nutr(�й��ٴ�Ӫ����־), 2003,11(2): 88-91. LUO X Y, LI Y Q, ZOU X L, et al.Determination of glutamic acid and glutamine in serum by HPLC with 2,4-dinitrochlorobenzene derivatization.Mod Prev Med(�ִ�Ԥ��ҽѧ), 2008,35(10): 1900-1902. LIU F N, LI J S, CHEN Y M, et al. Plasma glutamine level monitored with high performance liquid chromatography for patient infusing glutamine after small bowel transplantion. Chin J Chromatogr(ɫ��),1997,15(5): 445-446. LIANG J W, CHEN L, CAO L P, et al. The determination of GLN in plasma of burn patients by RP-HPLC.Hebei Med J(�ӱ�ҽҩ), 2003,25(8): 566-567. TEERLINK T, HENNEKES M W T, VAN LEEUWEN P A M, et al. Rapid determination of glutamine in biological samples by high-performance liquid chromatography. Clinchimacta, 1993, 218(2): 159-168. VAN DE MERBEL N C. Quantitative determination of endogenous compounds in biological samples using chromatographic techniques. TrAC Trends Anal Chem, 2008, 27(10): 924-933. ZHANG X, XIE X Q, LIU T L,et al.Quantative determination and bioequivalence assesement of the drugs as endogenous compounds.Chin J New Drugs(�й���ҩ��־), 2011,20(22): 2221-2228. European Medicines Agency. Committee for medicinal products for human use guideline on the investigation of bioequivalence(draft) .2008: 13. HUANG J H, HUANG X H, CHEN Z Y,et al.Dose conversion among different animals and healthy volunteers in pharmacological study.Chin J Clin Pharmacol Ther(�й��ٴ�ҩ��ѧ������ѧ), 2004,9(9): 1069-1072.