School of Chinese Materia Medica, Beijing University of Chinese Medicine, No.6, Zhonghuan South Road, Wangjing, Chaoyang district, Beijing 100102, China.
Molecules. 2010 May 7;15(5):3356-65. doi: 10.3390/molecules15053356.
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.
过敏是一种高度复杂的疾病，临床表现从轻度口腔、胃肠道、反复喘息和皮肤症状到危及生命的全身性疾病不等。花生四烯酸、类二十烷酸、组胺、有机酸和缬氨酸的水平被认为与过敏有关，具有多种生理功能。在这项研究中，我们开发了一种 RP-LC/MS 方法，用于分离和定量六种不同的潜在内源性生物标志物，包括白三烯 B(4) (LTB(4))、前列腺素 D(2) (PGD(2))、花生四烯酸 (AA)、组胺 (HI)、乳酸 (LA)和缬氨酸 (VAL)，从卵清蛋白 (OVA) 诱导过敏的大鼠和正常大鼠的血清中，并比较了模型组和对照组之间的差异。分离在 Prevail C18 柱（250mmx4.6mm，5μm）上进行，采用乙腈与 0.1%甲酸（v/v）和 10mM 甲酸铵（用甲酸调至 pH4.0）的梯度洗脱，流速为 0.5mLmin(-1)。该方法经过验证，具有较高的灵敏度、准确性（回收率值为 76.16-92.57%）和精密度（所有化合物的 RSD<10%），线性范围跨越多个数量级。该方法成功应用于大鼠血清，并表明能够指示大鼠体内生物标志物的内源性水平。对生物标志物的分析可以深入了解与相关疾病相关的过敏机制。
