College of Food Science and Engineering, Northwest A&F University, Yangling, China.
PLoS One. 2018 Mar 1;13(3):e0192964. doi: 10.1371/journal.pone.0192964. eCollection 2018.
This study aimed to analyze the bile acid patterns in commercially available oxgall powders used for evaluation of the bile tolerance ability of probiotic bacteria. Qxgall powders purchased from Sigma-Aldrich, Oxoid and BD Difco were dissolved in distilled water, and analyzed. Conjugated bile acids were profiled by ion-pair high-performance liquid chromatography (HPLC), free bile acids were detected as their p-bromophenacyl ester derivatives using reversed-phase HPLC after extraction with acetic ether, and total bile acids were analyzed by enzymatic-colorimetric assay. The results showed that 9 individual bile acids (i.e., taurocholic acid, glycocholic acid, taurodeoxycholic acid, glycodeoxycholic acid, taurochenodeoxycholic acid, glycochenodeoxycholic acid, cholic acid, chenodeoxycholic acid, deoxycholic acid) were present in each of the oxgall powders tested. The content of total bile acid among the three oxgall powders was similar; however, the relative contents of the individual bile acids among these oxgall powders were significantly different (P < 0.001). The oxgall powder from Sigma-Aldrich was closer to human bile in the ratios of glycine-conjugated bile acids to taurine-conjugated bile acids, dihydroxy bile acids to trihydroxy bile acids, and free bile acids to conjugated bile acids than the other powders were. It was concluded that the oxgall powder from Sigma-Aldrich should be used instead of those from Oxoid and BD Difco to evaluate the bile tolerance ability of probiotic bacteria as human bile model.
本研究旨在分析用于评估益生菌胆汁耐受能力的市售牛胆粉中的胆汁酸模式。从 Sigma-Aldrich、Oxoid 和 BD Difco 购买的 Qxgall 粉末溶解在蒸馏水中并进行分析。采用离子对高效液相色谱法(HPLC)对结合型胆汁酸进行分析,游离胆汁酸用乙酸乙酯提取后作为其对溴苯甲酰基酯衍生物,用反相 HPLC 检测,总胆汁酸用酶比色法分析。结果表明,9 种游离型胆汁酸(即牛胆酸、甘氨胆酸、牛脱氧胆酸、甘氨脱氧胆酸、牛磺鹅脱氧胆酸、甘氨鹅脱氧胆酸、胆酸、鹅脱氧胆酸、脱氧胆酸)存在于每种牛胆粉中。三种牛胆粉中的总胆汁酸含量相似,但这些牛胆粉中各胆汁酸的相对含量差异显著(P<0.001)。与其他两种牛胆粉相比,Sigma-Aldrich 牛胆粉中的甘氨酸结合型胆汁酸与牛磺酸结合型胆汁酸、二羟基胆汁酸与三羟基胆汁酸以及游离胆汁酸与结合型胆汁酸的比例更接近人胆汁。因此,建议使用 Sigma-Aldrich 牛胆粉代替 Oxoid 和 BD Difco 牛胆粉作为人胆汁模型来评估益生菌的胆汁耐受能力。