A LC-MS-based method for quantification of biomarkers from serum of allergic rats.
Allergies are highly complex disorders with clinical manifestations ranging from mild oral, gastrointestinal, recurrent wheezing, and cutaneous symptoms to life-threatening systemic conditions. The levels of arachidonic acid, eicosanoids, histamine, organic acids and valine are considered to have a variety of physiological functions in connection with allergies. In this research, we have developed a RP-LC/MS method to separate and quantitate six different potential endogenous biomarkers, including leukotriene B(4) (LTB(4)), prostaglandin D(2) (PGD(2)), arachidonic acid (AA), histamine (HI), lactic acid (LA) and valine (VAL), from serum of rats with ovalbumin (OVA)-induced allergy and normal rats, and the discrepancies between the model group and the control group were compared. The separation was performed on a Prevail C18 column (250 mm x 4.6 mm, 5 microm) with a gradient elution of acetonitrile with 0.1% formic acid (v/v) and 10 mM ammonium formate (adjusted to pH 4.0 with formic acid) at a flow rate of 0.5 mL min(-1) The method was validated and shown to be sensitive, accurate (recovery values 76.16-92.57%) and precise (RSD < 10% for all compounds) with a linear range over several orders of magnitude. The method was successfully applied to rat serum and shown to be indicative of the endogenous levels of biomarkers within the rat body. The analysis of the biomarkers can provide insight into the allergic mechanisms associated with related diseases.
Bai
Yu Jing
YJ
School of Chinese Materia Medica, Beijing University of Chinese Medicine, No.6, Zhonghuan South Road, Wangjing, Chaoyang district, Beijing 100102, China. jingjing8371332@sina.com
Gao
Xiao Yan
XY
Lu
Jian Qiu
JQ
Zhang
Hong Gui
HG
D000818:Animals; D016718:Arachidonic Acid / Q000097:blood; D015415:Biomarkers / Q000097:blood; D002853:Chromatography, Liquid / Q000379:methods; D006632:Histamine / Q000097:blood; D006967:Hypersensitivity / Q000097:blood / Q000175:diagnosis* / Q000209:etiology; D019344:Lactic Acid / Q000097:blood; D007975:Leukotriene B4 / Q000097:blood; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards; D010047:Ovalbumin; D015230:Prostaglandin D2 / Q000097:blood; D051381:Rats; D012680:Sensitivity and Specificity; D014633:Valine / Q000097:blood
D015415:Biomarkers; D007975:Leukotriene B4; D016718:Arachidonic Acid; D019344:Lactic Acid; D006632:Histamine; D010047:Ovalbumin; D014633:Valine; D015230:Prostaglandin D2