Rapid small intestinal permeability assay based on riboflavin and lactulose detected by bis-boronic acid appended benzyl viologens.
Rapid small intestinal permeability assay based on riboflavin and lactulose detected by bis-boronic acid appended benzyl viologens.
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基于核黄素和乳糖的快速小肠渗透性测定，由双硼酸附加苄基紫胶检测。
DOI：
10.1016/j.cca.2014.09.031
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发表时间：
2015-01-15
期刊：
影响因子：
--
通讯作者：
Webb DL
中科院分区：
文献类型：
--
作者：
Resendez A;Abdul Halim M;Landhage CM;Hellström PM;Singaram B;Webb DL
虽然有机硼酸是有效的高通量糖传感器，但它们尚未被用于肠道渗透性研究。描述了乳果糖/甘露醇测定的改进，通过该改进，在尿液收集时使用乳果糖/核黄素比率评估小肠通透性。志愿者摄入50毫克核黄素和5克甘露醇或10克乳果糖。收集尿液6小时。用自体荧光法测定核黄素。采用C18固相萃取法去除核黄素。然后使用与荧光团HPTS偶联的1，1 ′-双（2-硼苄基）-4，4 ′-联吡啶鎓（4，4 ′oBBV）测定乳果糖和甘露醇。核黄素与甘露醇在6小时内的时间曲线。尿液中的核黄素回收率为11.1 ± 1.9%（平均值± SEM，n = 7），与甘露醇相似。4，4 ′oBBV与乳果糖有选择性结合，可能涉及果糖和半乳糖部分之间的协同作用。检测和定量下限为90和364 μM。乳果糖测定对其他渗透性探针不敏感（例如，蔗糖、三氯蔗糖），同时耐受葡萄糖或乳糖。该测定可适用于自动化系统。4，4 ′oBBV的稳定性超过4年。通过自体荧光测量的核黄素与用4，4 ′oBBV测量的乳果糖相结合，代表了一种用于快速测量肠道通透性的有用的新化学，具有优异的稳定性、成本和通量优势。
Although organoboronic acids are efficient high-throughput sugar sensors, they have not been pursued for gut permeability studies. A modification of the lactulose/mannitol assay is described by which small intestinal permeability is assessed at the time of urine collection using a lactulose/riboflavin ratio. Volunteers ingested 50 mg riboflavin and either 5 g mannitol or 10 g lactulose. Urine was collected for 6 hrs. Riboflavin was assayed by autofluorescence. Riboflavin was removed by C18 solid phase extraction. Lactulose and mannitol were then assayed using 1,1′-bis(2-boronobenzyl)-4,4′-bipyridinium (4,4′oBBV) coupled to the fluorophore HPTS. The temporal profile over 6 hrs for riboflavin paralleled mannitol. Riboflavin recovery in urine was 11.1 ± 1.9 % (mean ± SEM, n = 7), similar to mannitol. There was selective binding of 4,4′oBBV to lactulose, likely involving cooperativity between the fructose and galactose moieties. Lower limits of detection and quantification were 90 and 364 μM. The lactulose assay was insensitive to other permeability probes (e.g., sucrose, sucralose) while tolerating glucose or lactose. This assay can be adapted to automated systems. Stability of 4,4′oBBV exceeds 4 years. Riboflavin measured by autofluorescence combined with lactulose measured with 4,4′oBBV represents a useful new chemistry for rapid measurement of intestinal permeability with excellent stability, cost and throughput benefits.
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影响因子：
5
作者：
NORTHROP, CA;LUNN, PG;BEHRENS, RH
通讯作者：
BEHRENS, RH
影响因子：
5
作者：
LUNN, PG;NORTHROP, CA;NORTHROP, AJ
通讯作者：
NORTHROP, AJ
影响因子：
3.4
作者：
Subramanian, Veedamali S.;Subramanya, Sandeep B.;Said, Hamid M.
通讯作者：
Said, Hamid M.
影响因子：
3.5
作者：
Camilleri M;Madsen K;Spiller R;Greenwood-Van Meerveld B;Verne GN
通讯作者：
Verne GN
影响因子：
1.8
作者：
Camara, JN;Suri, JT;Singaram, B
通讯作者：
Singaram, B