University of Florida, College of Veterinary Medicine, Department of Physiological Sciences, Florida Racing Laboratory, Gainesville, FL 32610-0117, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Mar 15;889-890:130-7. doi: 10.1016/j.jchromb.2012.02.008. Epub 2012 Feb 13.
We describe a validated, rapid, sensitive, and specific UHPLC-MS/MS method to detect and quantify glycopyrrolate in 0.5 mL of horse urine. Further, we investigated the elimination of glycopyrrolate in urine after both intravenous and oral administration of clinically relevant doses to Thoroughbred horses. Quantification was performed by weighted, linear regression analysis using a deuterated analogue of glycopyrrolate as internal standard (IS). The method was characterized by a linear range of 5-2500 pg/mL, a lower limit of quantification of 5 pg/mL and a limit of detection of 1 pg/mL. The intra and inter-batch imprecisions were <10% RSD and accuracy of the method ranged between 94 and 104%. Glycopyrrolate remained detectable in urine samples collected through 168 h after intravenous administration and through 24h after oral administration. Analytical method validation requirements for linearity, specificity, precision, accuracy, stability, dilution integrity, matrix effect, and ruggedness have been fulfilled. The urine method described in this report is simple and efficient and is the first reported method with sufficient sensitivity, accuracy, and precision to regulate the use of glycopyrrolate in urine samples collected more than one day after dosing of horses. Urine to plasma glycopyrrolate concentration ratios were calculated and were approximately 100:1 in samples collected from 24h through the end of sample collection.
我们描述了一种经过验证的、快速、灵敏和特异的 UHPLC-MS/MS 方法，用于检测和定量 0.5mL 马尿中的格隆溴铵。此外，我们还研究了在给纯种马静脉内和口服给予临床相关剂量后，格隆溴铵在尿液中的消除情况。通过使用格隆溴铵的氘代类似物作为内标（IS）进行加权线性回归分析来进行定量。该方法的线性范围为 5-2500pg/mL，定量下限为 5pg/mL，检测限为 1pg/mL。批内和批间精密度均<10%RSD，方法的准确度在 94%至 104%之间。静脉内给药后 168 小时内和口服给药后 24 小时内仍可在尿液样本中检测到格隆溴铵。已满足线性、特异性、精密度、准确度、稳定性、稀释完整性、基质效应和耐用性的分析方法验证要求。本报告中描述的尿液方法简单高效，是第一个具有足够灵敏度、准确度和精密度的方法，可用于监管在给马用药后一天以上采集的尿液样本中格隆溴铵的使用。计算了尿液与血浆中格隆溴铵浓度比，在 24 小时至样品采集结束时的样本中约为 100:1。
