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Chemical Research in Chinese Universities ›› 2011, Vol. 27 ›› Issue (4): 578-583.

• Articles • Previous Articles     Next Articles

Development of a Derivatization-free GC-FID Method for Evaluation of Free Fatty Acid Levels in Plasma of Diabetic Nephropathy Patients

HUA Hai-yin1, LIANG Qiong-lin1*, CHEN Jun1, WANG Yi-ming1, LI Ping2 and LUO Guo-an1*   

  1. 1. Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, Ministry of Education, Department of Chemistry, Tsinghua University, Beijing 100084, P. R. China;
    2. Institute of Clinical Medical Science, China-Japan Friendship Hospital, Beijing 100029, P. R. China
  • Received:2010-05-28 Revised:2010-08-10 Online:2011-07-25 Published:2011-06-29
  • Contact: LUO Guo-an;LIANG Qiong-lin E-mail:luoga@mail.tsinghua.edu.cn;liangql@mail.tsinghua.edu.cn
  • Supported by:

    Supported by the National Basic Research Program of China(Nos.2007CB511903, 2005CB523503), the International Co- operation Project of Ministry of Science and Technology of China(No.S2010GR0583) and the National Natural Science Foundation of China(Nos.90709045, 20805026).

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Abstract: Metabolism of free fatty acids(FFAs) is related to several important physiological events and therefore their quantitaion in biological samples arouses extensive interest and efforts. Existing gas chromatography with flame ionization detector(GC-FID) methods for the analysis of FFAs normally require derivatization of them in order to lower boiling points. But this extra procedure tends to induce additional error and it is laborious and time-consuming. A derivatization-free method was therefore established in the present investigation to determine FFAs in human plasma by capillary(GC-FID). After extraction of FFAs from plasma, a highly polar FFAP(free fatty acid in plasma) column was employed to directly quantitate FFAs concentration, free from derivatization reaction. All sample pretreatments were carried out at room temperature, improving recovery of short-chain FFAs. Heptadecanoic acid(C17:0) was employed as internal standard, and the proposed method was validated for recovery, precision, sensitivity, stabi- lity, and linearity. Validation data show that it is suitable for clinical study that has been applied to the evaluation of FFAs levels in plasma of diabetic nephropathy(DN) patients during the course of treatment. Forty-seven patients diagnosed with DN were admitted to the double-blind experiment. Control group(n=17) underwent solely basic treatment and the patients did not show significant change in FFAs concentration during six months of treatment. Experiment group(n=30) was supplied with traditional Chinese medicine besides basic treatment. After six months of medication, their plasma concentration of palmitic acid(C16:0), stearic acid(C18:0) and oleic acid(C18:1n–9) decreased while linolenic acid(C18:3n–3) increased significantly(P<0.05). These four compounds could be served as biomar- kers in the evaluation of drug efficacy, and their quantitation in plasma may provide additional information for   disease progression in DN patients.

Key words: Derivitizaiton-free gas chromatography with flame ionization detector(GC-FID), Free fatty acid(FFA), Diabetic nephropathy(DN)