Department of Oriental Pharmaceutical Sciences, Kyung Hee East-West Pharmaceutical Research Institute, College of Pharmacy, Kyung Hee University, Hoegi-dong, Dongdaemoon-gu, Seoul 130-701, Republic of Korea.
J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Jul 1;930:70-4. doi: 10.1016/j.jchromb.2013.04.033. Epub 2013 Apr 29.
A method for determination of proline-hydroxyproline dipeptide (PHP) was developed using high-performance anion-exchange chromatography-pulsed amperometric detection (HPAEC-PAD). This method resulted in good separation of proline (Pro), hydroxyproline (Hyp), and PHP within 20min using a mobile phase of 1.2mM Ba(OH)2+1.5mM Ba(OAc)2. The linear dynamic ranges and their detection limits (S/N=3) were 1-100 (r(2)=0.9990-0.9999) and 0.05-0.3μM, respectively. Mean recoveries were 91.6-121.3% and 92.2-110.3% for intra- and inter-day assays, respectively. Our HPAEC-PAD method showed clear differences in the corrected PHP levels measured in urine samples from two groups of rats, sham-operated and ovariectomized, without the need for prior acid hydrolysis or sample derivatization.
建立了一种采用高效阴离子交换色谱-脉冲安培检测（HPAEC-PAD）测定脯氨酸-羟脯氨酸二肽（PHP）的方法。该方法在 20min 内使用 1.2mM Ba(OH)2+1.5mM Ba(OAc)2 作为流动相，可实现脯氨酸（Pro）、羟脯氨酸（Hyp）和 PHP 的良好分离。线性动态范围及其检测限（S/N=3）分别为 1-100（r(2)=0.9990-0.9999）和 0.05-0.3μM。对于日内和日间测定，平均回收率分别为 91.6-121.3%和 92.2-110.3%。我们的 HPAEC-PAD 方法无需进行酸水解或样品衍生化，即可在两组大鼠（假手术组和卵巢切除组）的尿液样本中测量到明显不同的校正 PHP 水平。
