目的 建立氣相色譜-質(zhì)譜聯(lián)用（GC/MS）的短鏈脂肪酸（SCFAs）分析方法，分析腸炎小鼠SCFAs代謝情況。方法 鹽酸飽和氯化鈉溶液酸化生物樣本，醋酸乙酯提取SCFAs后衍生化，建立GC/MS方法進(jìn)行生物樣本中SCFAs測定；考察鹽酸溶液提取和鹽酸飽和氯化鈉溶液提取SCFAs總離子流圖譜；考察SCFAs混合物衍生化前后的穩(wěn)定性，糞便、血清、小腸、結(jié)腸、肝臟組織中SCFAs測定方法的線性、精密度及準(zhǔn)確度、提取回收率和基質(zhì)效應(yīng)。運(yùn)用建立的方法對正常及硫酸葡聚糖（DSS）慢性腸炎小鼠血、肝、小腸、結(jié)腸、結(jié)腸內(nèi)容物以及盲腸內(nèi)容物中發(fā)揮重要作用的SCFAs進(jìn)行測定。結(jié)果 用鹽酸飽和氯化鈉溶液進(jìn)行SCFAs提取后，建立的GC/MS法可同時測定甲酸、乙酸、丙酸、異丁酸、丁酸、異戊酸、戊酸、異己酸、己酸、乳酸、琥珀酸共11種SCFAs；方法學(xué)考證顯示，該方法具有較好的準(zhǔn)確度、精密度以及提取回收率；除乙酸外，其余SCFAs基質(zhì)效應(yīng)均較好，不能完全除去乙酸的基質(zhì)效應(yīng)，但其RSD均小于3.8%，說明基質(zhì)的影響效應(yīng)相對穩(wěn)定，對結(jié)果準(zhǔn)確性影響較小。與對照組比較，慢性腸炎小鼠結(jié)腸內(nèi)容物SCFAs蓄積；盲腸內(nèi)容物SCFAs整體下調(diào)，乳酸蓄積，提示腸炎小鼠SCFAs代謝紊亂。結(jié)論 建立了生物樣品中SCFAs提取和檢測方法，并應(yīng)用于慢性腸炎小鼠SCFAs代謝分析，腸炎小鼠SCFAs代謝紊亂。

Objective To analyze the metabolism of short chain fatty acids (SCFAs) in enteritis mice by gas chromatography-mass spectrometry (GC/MS). Method The pretreatment of biological samples contains acidification in sodium chloride solution saturated with hydrochloric acid, ethyl acetate extraction, and derivatization. GC/MS method is established to quantify SCFAs in biological samples. Total ion chromatograms of SCFAs of fecal samples extracted by hydrochloric acid solution and hydrochloric acid saturated sodium chloride solution were observed. The stability of the SCFAs mixture before and after the derivatization, the linearity, precision, accuracy, recovery, and the matrix effect of SCFAs determination in feces, sera, small intestine, colon, and liver tissues were investigated. The established method was used to determine the important SCFAs in the blood, liver, small intestine, colon, colon content, and the content of cecum in normal and DSS chronic enteritis mice. Results The method could quantify 11 different kinds of SCFAs including formic acid, acetic acid, propionic acid, isobutyric acid, butyric acid, isovalerate, valerate, ISO caproic acid, hexanoic acid, lactic acid, and succinic acid in biological samples extracted by hydrochloric acid saturated sodium chloride solution with great accuracy, precision, and extraction recovery. Except for acetic acid, the other SCFAs matrix effects were all good, and the matrix effect of acetic acid could not be completely removed, but the RSD was less than 3.8%, indicating that the effect of matrix was relatively stable and had little effect on the accuracy of the results. The method has been used to analyze SCFAs in mouse intestinal contents, liver, small intestinal, and colon. Compared with control group, colitis mouse showed SCFAs metabolic disorder caused by SCFAs accumulation in colon content and SCFAs reduction except for lactic acid in cecum content. Conclusion The SCFAs extraction and quantification method has been established and applied to analyze SCFAs in colitis mouse, and colitis mouse showed SCFAs metabolic disorder.

國家自然科學(xué)基金資助項(xiàng)目（81430091）