为制备地昔尼尔乳剂并建立其质量浓度的测定方法,以稳定常数和离心稳定性作为乳剂稳定性评价指标,采用单因素试验和正交设计试验法考察处方因素和工艺因素对乳剂稳定性的影响,以筛选出乳剂的最优处方和制备工艺;采用高效液相色谱法(HPLC)测定乳剂的药物质量浓度.结果显示,地昔尼尔乳剂的最优处方为地昔尼尔50 g/L、乳化剂70 mL/L、油相125 mL/L、助乳化剂160 mL/L、稳定剂为3.5 g/L;地昔尼尔乳剂的制备方法为乳化温度50℃,20 000 r/min高速搅拌10 min.建立的HPLC色谱条件为色谱柱X Bridge C18(4.6mm×250 mm,5μm),流动相乙腈与水(6 mL/L三乙胺+2 mL/L甲酸)的体积比为10∶90,流速0.8 mL/min,柱温30℃,紫外检测波长267 nm,进样体积20 μL.在10～100 μg/mL的质量浓度范围内,峰面积与药物浓度呈良好线性关系(r2 =0.999 9,n=7),回归方程为y=148.01x+91.629.精密度试验相对标准偏差均小于2％,在80％、100％和120％3个添加水平时,平均回收率为98.36％,精密度、回收率均符合要求.定量限和检测限分别为10 ng/mL和20 ng/mL,测得3批地昔尼尔乳剂的平均质量浓度为49.29 mg/mL,为标示量的98.57％.结果表明,该处方合理、制备工艺可靠,所建立的高效液相色谱法可用于地昔尼尔乳剂的检测.

The adjuvant activity of GLP was investigated in vitro and in vivo. In vitro experiment, the effects of GLP on chicken peripheral lymphocytes proliferation were compared by MTT assay. The results showed that GLP could significantly enhance lymphocytes proliferation singly or synergistically with ConA. The interferon-gamma (IFN-γ) mRNA levels of chicken peripheral lymphocytes stimulated by GLP synergistically with ConA were measured using fluorescent quantitative PCR. The results showed that GLP could promote interferon-γ mRNA levels in peripheral lymphocytes. In vivo experiment, 175 14-day-old chickens were randomly divided into 7 groups. The chickens except blank control (BC) group were vaccinated with Newcastle disease vaccine, repeated vaccination at 28 days old. At the same time of the first vaccination, the chickens in experimental groups were orally administrated with 5 different doses of GLP respectively, whereas vaccination control (VC) and BC groups were treated with physiological saline, once a day for three successive days. On Day 7, 14, 21 and 28 after the first vaccination, the peripheral lymphocytes proliferation and serum ND antibody titer were determined. The results showed that GLP could significantly promote lymphocyte proliferation and enhance serum antibody titer. The results indicated that GLP may be a novel immunomodulator.