屋尘螨变应原制剂外观异常对其质量的影响

目的分析屋尘螨变应原制剂外观异常对其质量的影响。方法依据进口药品注册标准(JS20130104)对1批(T6469)屋尘螨变应原制剂进行全面质量检定,同时与检定合格样品(A3016、A3064及A3110批)进行相应比对,Western blot鉴定主要变应原组分,纳米粒径颗粒分析仪检测制剂中铝佐剂-蛋白吸附物的中值粒径(D50)。选取检定合格样品进行冷冻模拟验证,观察低温冷冻对制剂外观性状的影响。结果 T6469批制剂外观不均一(外观性状异常与正常样品),混悬液浊度有差异,部分样品出现摇不散的贴壁颗粒,外观性状异常样品与正常样品比较,浑浊度降低,沉降速度增快,铝佐剂吸附物液面高度明显低于外观正常样品。经检定,T6469、A3016、A3064及A3110批制剂pH、铝含量、苯酚含量及游离上清活性均在质量标准范围内,在相对分子质量25 000及14 000处均出现特异性条带,但T6469批外观性状异常样品条带变弱。T6469外观性状异常与正常样品之间D50差异有统计学意义(P <0. 05);检定合格样品冻存前后D50差异有统计学意义(P <0. 05)。T6469批外观性状异常样品与正常样品和检定合格样品比较,总生物学活性明显降低,差异有统计学意义(P <0. 05)。检定合格样品放置低温环境至冻结后重现了上述外观异常性状,生物活性与未处理样品相比显著降低(P <0. 05)。结论该批屋尘螨变应原制剂的外观异常可能是由于样品贮存及运输中出现低温冻结,引起铝佐剂-蛋白凝胶颗粒出现聚集,形成较大颗粒,最终导致其生物活性显著降低。

Influence of abnormal appearance on quality of house dust mite allergen products

Objective To analyze the abnormal appearance on quality of house dust mite allergen products. Methods Overall control tests were performed on one batch of house dust mite allergen product(T6469) according to the requirements for license of imported drugs(JS20130104),of which the results were compared with those of qualified samples(Lots A3016,A3064 and A3110). The components of major allergen was identified by Western blot,while the median diameter(D50)of aluminum adjuvant-protein adsorbate in the product by nanoparticle size analyzer. The samples qualified in the control tests were subjected to mimic freezing verification to observe the influence of freezing on appearance of the product. Results The appearance of product of lot T6469 was inhomogeneous,with different turbidity,and adherent particles not dispersed on shaking in partial samples. Compared with those with normal appearance,the turbidity of samples with abnormal appearance decreased,while the sedimentation rate increased,and the liquid level of adjuvant-protein adsorbate decreased. The pH value,aluminum content,phenol content and activity of free supernatant of products of lots T6469,A3016,A3064 and A3110 met the quality standard,while specific bands with relative molecular masses of 25 000 and 14 000 were observed. However,the bands of product of lot T6469,with abnormal appearance,were light in color. The D50 of product of lot T6469 with normal and abnormal appearances showed significant difference(P < 0. 05). Meanwhile, the D50 of samples qualified in control tests showed significant difference before and after storage in frozen(P < 0. 05). The total biological activity of product of lot T6469 with abnormal appearance was significantly lower than those of product with normal appearance and the qualified products as control(P < 0. 05). However,the above-mentioned abnormal features appeared again in the product qualified in control tests after freezing, while the biological activity decreased significantly as compared with that of samples without freezing(P < 0. 05). Conclusion The abnormal appearance of the batch of house dust mite allergen product might be due to the freezing during storage and transport, resulting the formation of large particles and significant decrease of biological activity.