气相色谱法同时测定复方庚酸炔诺酮注射液中的苯甲醇和苯甲醛

Simultaneous determination of benzyl alcohol and benzaldehyde in compound norethisterone enanthate injection by gas chromatograph

目的:建立气相色谱法同时测定复方庚酸炔诺酮注射液中苯甲醇与苯甲醛的含量,以控制复方庚酸炔诺酮注射液的质量。方法:采用毛细管柱气相色谱法,用聚乙二醇毛细管色谱柱(DB-WAX,30 m×0.53 mm,1.0μm),进样口温度为200℃,分流比为10∶1,恒压模式,柱压保持在34.5 kPa,初始柱温为50℃,以每分钟5℃的升温速率升至220℃,保持15min,载气为氦气,以FID为检测器,检测器温度为300℃,进样量为1μL。结果:苯甲醇在0.032～40.13 mg.mL-1范围内峰面积与浓度的线性关系良好,r为1.0000(n=7),高、中、低平均加样回收率分别为94.3%、94.3%、94.9%,RSD分别为0.9%、0.7%、2.6%(n=3);苯甲醛在0.001～0.7 mg.mL-1的范围内峰面积与浓度的线性关系良好,r为0.99999(n=7),高、中、低平均加样回收率分别为95.9%、96.4%、99.8%,RSD分别为0.6%、1.2%、3.3%(n=3)。苯甲醇的高、中、低浓度的日间精密度的RSD分别为0.8%、1.1%、1.7%(n=3);苯甲醛的高、中、低浓度的日间精密度的RSD分别为0.7%、0.9%、3.4%(n=3)。苯甲醇与苯甲醛定量限分别为1.9 ng(RSD=2.5%,n=10)和2.3 ng(RSD=4.6%,n=10),检测限分别为0.58 ng和0.69 ng;对照品溶液在96h内保持稳定。测定了4批不同批号的样品,结果苯甲醇的含量比较稳定,分别为1.97%、1.97%、2.02%和2.04%,与处方投样量(2%)保持一致,苯甲醛的含量因生产日期不同有很大的差异,分别为0.09%、0.08%(均为2008年生产)、0.01%、0.01%(均为2010年生产)。根据处方,苯甲醛主要由苯甲醇氧化而得,将结果折算成以样品中苯甲醇的量计,含苯甲醛分别为4.7%、4.3%、0.5%和0.4%。结论:本法操作简单,结果准确,专属性好,可用作控制复方庚酸炔诺酮注射液中苯甲醇的含量,同时可监测该制剂中苯甲醛的量的变化。在复方庚酸炔诺酮注射液贮存过程中,苯甲醛含量随贮存时间不断增加,可能会产生与苯甲醛相关的不良反应,应该纳入到监管检测的范围里来。而其他含苯甲醇的注射液中苯甲醛是否过量也应受到重视和研究。

Objective： To establish a GC method for the simultaneous determination of benzyl alcohol and benzal- dehyde in compound norethisterone enanthate injection. Methods： A capillary gas chromatographic method was car- fled out. An Agilent DB -WAX capillary column（30 m ×0.53 ram,1.0 μm）was used with the injection port tem- perature of 200℃, and the split ratio of 10 ： 1. A constant pressure at 34.5 kPa was set up for the column. The ini- tial column temperature was 50℃ ,to 220 ℃ at 5 ℃ · rain^-1 ramp and hold for 15 rain. The temperature of FID was set at 300 ℃, and the sample volume was set at 1 μL. Results： The peak area and concentration of benzyl alco- hol showed a good linearity within the concentration range of 0. 032 -40. 13 mg·mL^-l（r = 1. 0000,n =7）. The high,medium and low recoveries of benzyl alcohol were 94.3% , 94.3% , and 94.9% （ RSD 0. 9% , 0. 7% and 2.6% , n = 3 ）, respectively ; The peak area and concentration of benzaldehyde showed a good linearity within the concentration range of 0. 001 - 0.7 mg~ mL-1 （ r = 0. 99999, n = 7）. The h;gh, medium and low recoveries of ben- zaldehyde were 95.9% ,96.4% and 99. 8% （ RSD = 0.6%, 1.2% and 3.3%, n = 3 ）, respectively. The inter - day precisions were O. 8% , 1.1% ,and 1.7% （n = 3 ）for hizh. medium and low concentrations of henzvl alcohol re,nee-tively ; The inter - day precisions were 0.7 % , 0.9 % and 3.4 % （ n = 3 ） for high, medium and low concentrations of benzaldehyde ;The LOQs of benzyl alcohol and benzaldehyde were 1.9 ng（ RSD = 2.5%, n = 10）and 2.3 ng（ RSD =4. 6% ,n = 10） ,respectively,while the LODs of benzyl alcohol and benzaldehyde were 0.58 ng and 0. 69 ng,re- speetively. The reference standard solution was stable within 96 hours. The testing result of and In total, four differ- ent batches of samples were tested, in whieh, the test results for benzyl aleohol were more stable - - 1.97%, 1.97% ,2.02%, and 2.04%, accorded with the prescription sampling volume; in comparison, the test results for benzaldehyde were mueh different between eaeh batch due to different produetion dates - - 0. 09% , and 0.08% （manufactured in 2008 ） , as well as 0. 01% , and 0. 01% （manufaetured in 2010）. According to prescriptions, since benzaldehyde was mainly derived from benzyl aleohol in compound norethisterone enanthate injection, and the a- mount of benzaldehyde was recalculated based on the amount of benzyl aleohol in the injection,the amount of ben- zaldehyde was ealeulated as 4.7% ,4.3% ,0.5%, and 0. 4%. Conclusion： The method is simple, aecurate, relia- ble, and can be used for quality eontrol of benzyl alcohol in compound norethisterone enanthate injection. This meth- od ean also be utilized to monitor the content variation of benzaldehyde in eompound norethisterone enanthate injec- tion at the same time. The amount of benzaldehyde may cause adverse drug reactions since it increases continually in the process of storage of compound norethisterone enanthate injection,thus, benzaldehyde test should be included in the inspeetion scope. Researehers and supervising organizations should also pay more attention to the amount of ben- zaldehyde in other injeetions eontaining benzyl alcohol.