Alvarez Rafael Herrera, Natal Fabio Luis Nogueira, Ribela Maria Teresa Carvalho Pinto, de Almeida Beatriz Elane, de Oliveira João Ezequiel, Bartolini Paolo
Agency for Agribusiness Technology of São Paulo, 13400-970 Piracicaba, Brazil.
Biotechnology Department, IPEN-CNEN, Cidade Universitária Sao Paulo, 05508-000 São Paulo, Brazil.
J Vet Sci. 2016 Dec 30;17(4):459-465. doi: 10.4142/jvs.2016.17.4.459.
Ovarian stimulation with commercial preparations of equine chorionic gonadotropin (eCG) produces extremely variable responses in domestic animals, ranging from excessive stimulation to practically no stimulation, when applied on the basis of their declared unitage. This study was conducted to analyze four commercial preparations from different manufacturers via reversed-phase HPLC (RP-HPLC) in comparison with a reference preparation and an official International Standard from the World Health Organization. The peaks obtained by this qualitative and quantitative physical-chemical analysis were compared using an bioassay based on the ovarian weight gain of prepubertal female rats. The RP-HPLC data showed one or two peaks close to a main peak (t= 27.9 min), which were related to the bioactivity. Commercial preparations that have this altered peak showed very little or no activity, as demonstrated by rat ovarian weight and in peripubertal gilts induced to ovulate. Overall, these findings indicate that RP-HPLC can be a rapid and reliable tool to reveal changes in the physicochemical profile of commercial eCG that is apparently related to decreased biological activity of this hormone.
使用马绒毛膜促性腺激素(eCG)的商业制剂对家畜进行卵巢刺激时,根据其宣称的单位剂量应用,会产生极其多变的反应,从过度刺激到几乎没有刺激。本研究通过反相高效液相色谱(RP-HPLC)分析了来自不同制造商的四种商业制剂,并与一种参比制剂和世界卫生组织的官方国际标准进行了比较。通过基于青春期前雌性大鼠卵巢增重的生物测定法,对这种定性和定量物理化学分析获得的峰进行了比较。RP-HPLC数据显示,靠近主峰(t = 27.9分钟)有一个或两个峰,这些峰与生物活性相关。如大鼠卵巢重量以及诱导排卵的青春期前后备母猪所示,具有这种改变峰的商业制剂显示出很少或没有活性。总体而言,这些发现表明,RP-HPLC可以作为一种快速可靠的工具,来揭示商业eCG物理化学特征的变化,这种变化显然与该激素生物活性的降低有关。