School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, People's Republic of China.
Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, Qingdao 266003, People's Republic of China.
Anal Methods. 2022 Jun 9;14(22):2227-2234. doi: 10.1039/d2ay00700b.
A sensitive and robust ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established for the first time to simultaneously quantify marine neutral neoagaro-oligosaccharides (NAOS) and agar-oligosaccharides (AOS) with different degrees of polymerization (DP) in Hanks' balanced salt solution (HBSS). The separation was achieved on a BEH amide column using a mobile phase of acetonitrile-10 mmol L ammonium acetate (58 : 42, v/v) with an isocratic elution program. The total analysis time was 3.5 min. The mass spectra were acquired in the multiple reaction monitoring (MRM) pattern by using a heated-electrospray ionization (H-ESI) source operating in the positive ionization mode. The linear range was 40-20 000 nmol L. The accuracy and precision ranged from 91.5 to 110.0% and 0.9 to 10.4%, respectively. The extraction recovery was consistent and reproducible. The stability was within 90.3-110.8%. The matrix effect, carryover, and dilution integrity were all satisfactory. Moreover, the validated method was successfully applied to the intestinal transport study by using the Caco-2 cell monolayer . The results revealed that neoagarobiose, neoagarotetraose, neoagarohexaose, agarotriose, agaropentose, and agaroheptose were transported by a paracellular pathway.
首次建立了一种灵敏、稳健的超高效液相色谱-串联质谱（UHPLC-MS/MS）方法，用于同时定量测定 Hank's 平衡盐溶液（HBSS）中不同聚合度（DP）的海洋中性新琼寡糖（NAOS）和琼脂寡糖（AOS）。采用乙腈-10 mmol L 乙酸铵（58：42，v/v）作为流动相，在 BEH 酰胺柱上进行分离，采用等度洗脱程序。总分析时间为 3.5 分钟。通过使用正离子模式下工作的加热电喷雾电离（H-ESI）源，在多重反应监测（MRM）模式下获取质谱。线性范围为 40-20000 nmol L。准确度和精密度范围分别为 91.5-110.0%和 0.9-10.4%。提取回收率一致且重现性好。稳定性在 90.3-110.8%之间。基质效应、残留和稀释完整性均令人满意。此外，该经过验证的方法成功应用于 Caco-2 细胞单层的肠道转运研究。结果表明，新琼二糖、新琼四糖、新琼六糖、琼脂三糖、琼脂五糖和琼脂七糖通过细胞旁途径转运。
