HPLC法测定肝素钠注射液中6种抑菌剂

目的 建立高效液相色谱（HPLC）法同时测定肝素钠注射液中苯酚、苯甲醇、三氯叔丁醇、羟苯甲酯、羟苯乙酯、羟苯丙酯6种抑菌剂。方法 采用Agilent Zorbax SB-C₁₈（4.6 mm×250 mm,5 μm）色谱柱,流动相A为水,流动相B为甲醇,梯度洗脱,流速为1.0 mL·min^-1,检测波长为220 nm及256 nm,柱温30 ℃。结果 苯酚、苯甲醇、羟苯甲酯、羟苯乙酯、三氯叔丁醇、羟苯丙酯分别在25.00~400.06 μg·mL^-1（r=1.000 0）、104.49~1 671.88 μg·mL^-1（r=0.999 9）、30.65~490.4 μg·mL^-1（r=0.999 9）、5.36~85.81 μg·mL^-1（r=0.999 9）、49.60~793.54 μg·mL^-1（r=0.999 9）、5.03~80.45 μg·mL^-1（r=1.000 0）内线性关系良好,平均回收率分别为100.3%、99.7%、99.8%、100.0%、99.4%和100.1%,RSD分别为1.0%、1.2%、1.3%、1.4%、1.4%和1.4%（n=9）。结论 方法准确,重现性好,结果可靠,可用于肝素钠注射液中6种抑菌剂的测定。     

HPLC同时测定双氯芬酸钠滴眼液中3种抑菌剂的含量

目的 建立HPLC同时测定双氯芬酸钠滴眼液中羟苯乙酯、硫柳汞和苯扎氯铵含量的方法。方法 用十八烷基键合硅胶为填充剂,以1%三乙胺溶液（磷酸调节pH值至3.0）为流动相A,以甲醇为流动相B,进行梯度洗脱;流速1.0 mL·min^-1,柱温40℃,检测波长254 nm。结果 羟苯乙酯在20.58~205.8 μg·mL^-1、硫柳汞在8.242~82.42 μg·mL^-1、苯扎氯铵n-C₁₂H₂₅取代物在12.88~128.8 μg·mL^-1、苯扎氯铵n-C₁₄H₂₉取代物在6.624~66.24 μg·mL^-1内线性良好（r≥0.999 8）,平均回收率为99.3%~102.5%（n=9）。结论 该方法简单、准确、重复性好,可用于控制双氯芬酸钠滴眼液中羟苯乙酯、硫柳汞和苯扎氯铵的含量。     

依诺沙星滴眼液抑菌剂考察

目的 建立测定依诺沙星滴眼液中抑菌剂硫柳汞、羟苯乙酯和苯扎溴铵的HPLC含量测定方法,并对这3种抑菌剂的稳定性进行考察。方法 选用C₁₈柱,以1%三乙胺溶液（用磷酸调节pH值至3.0）为流动相A,以甲醇为流动相B,进行梯度洗脱;检测波长苯扎溴铵为218 nm,羟苯乙酯和硫柳汞为262 nm,进样量为20 μL。同时考察抑菌剂的稳定性。结果 羟苯乙酯、硫柳汞和苯扎溴铵之间的分离度分别为19.7,48.7。硫柳汞在9.1~151.4 μg·mL^-1、羟苯乙酯在2.1~41.2 μg·mL^-1、苯扎溴铵在21.8~218.0 μg·mL^-1内均呈现良好的线性关系（r=0.999,1.000,0.999）,其检出限分别为1.4,1.2,13.1 ng。在高温条件下,硫柳汞和羟苯乙酯含量均略有下降;在紫外光照条件下,硫柳汞含量下降明显;苯扎溴铵几乎不受高温和紫外光照的影响。结论 该方法灵敏度高,能够有效评估滴眼液中抑菌剂的含量。     

HPLC法同时测定吡诺克辛钠滴眼液中不同抑菌剂的含量

目的：建立同时测定吡诺克辛钠滴眼液中不同抑菌剂（硫柳汞钠、羟苯甲酯、羟苯乙酯、羟苯丙酯）含量的HPLC法。方法：采用C18色谱柱（4.6 mm×150 mm,5 μm）,流动相为0.005 mol·L^-1的醋酸铵溶液（每1000 mL中含三乙胺10 mL,用冰醋酸调节pH值至5.0±0.5）-乙腈（70：30）,流速为1.0 mL·min^-1,检测波长为256 nm,柱温为30℃。结果：硫柳汞钠、羟苯甲酯、羟苯乙酯和羟苯丙酯在各自的检测质量浓度范围内线性关系良好,r为0.9993~1.0000,检测限分别为2.3、0.4、0.7、1.1 ng;4种抑菌剂的平均回收率为100.4%~102.7%（RSD≤1.4,n=9）。结论：本文建立的方法结果准确可靠,可作为吡诺克辛钠滴眼液中不同抑菌剂的质量控制方法。     

HPLC法检测醋酸奥曲肽注射剂中5种抑菌剂

目的：检测醋酸奥曲肽注射剂中是否添加苯酚、苯甲醇、三氯叔丁醇、对羟基苯甲酸甲酯及对羟基苯甲酸丙酯5种抑菌剂。方法：采用C18（L）（4.6 mm×250 mm,5 μm）色谱柱,流动相A为水,流动相B为甲醇,梯度洗脱,流速为1.0 mL·min^-1,柱温30℃,检测波长为220 nm及256 nm。结果：苯酚、苯甲醇、三氯叔丁醇、对羟基苯甲酸甲酯及对羟基苯甲酸丙酯各峰均分离良好;苯酚在1.29~258.60 μg·mL^-1范围内线性关系良好（r=1.000）,平均回收率为100.0%（n=9）;苯甲醇在2.72~544.20 μg·mL^-1范围内线性关系良好（r=0.9997）,平均回收率为100.3%（n=9）;三氯叔丁醇在9.95~1990.00 μg·mL^-1范围内线性关系良好（r=1.000）,平均回收率为100.9%（n=9）;对羟基苯甲酸甲酯在0.27~53.80 μg·mL^-1范围内线性关系良好（r=1.000）,平均回收率为100.5%（n=9）;对羟基苯甲酸丙酯在0.26~52.00 μg·mL^-1范围内线性关系良好（r=1.000）,平均回收率为99.4%（n=9）;91批醋酸奥曲肽注射剂中均未检出5种抑菌剂。结论：该方法准确、灵敏、简便,可用于醋酸奥曲肽注射剂中苯酚、苯甲醇、三氯叔丁醇、对羟基苯甲酸甲酯及对羟基苯甲酸丙酯5种抑菌剂的检查。     

HPLC法测定黄连上清片中连翘酯苷A的含量

目的 采用高效液相色谱法测定黄连上清片中连翘酯苷A的含量。方法 色谱柱：VP-ODSC₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相：乙腈-0.4%冰醋酸溶液（15:85）,流速1.0 mL·min^-1,检测波长330 nm。结果 连翘酯苷A在10.24 ~81.92 µg·mL^-1范围内与峰面积线性关系良好(r=0.999 8),精密度试验RSD为0.85%,重复性试验RSD为1.14%,平均回收率为97.91%,RSD为1.55%(n=6)。结论 该方法操作简单,精密度良好,结果准确可靠,适用于黄连上清片中连翘酯苷A的含量测定。     

HPLC法测定化妆品原料2-羟乙基脲及杂质尿素的含量

目的 建立化妆品原料2-羟乙基脲的高效液相色谱（high performance liquid chromatography,HPLC）含量测定方法,并检测其杂质尿素的含量,以实现对2-羟乙基脲的质量控制。方法采用Capcell PAK ADME C_{18色谱柱（4.6 mm×250 mm,5 μm）;以水为流动相;柱温30℃;流速为1.0 mL·min^-1;进样量10 μL;检测波长194 nm。结果 2-羟乙基脲和尿素分别在1.168～116.8 μg·mL^-1（r=0.999 7）和1.032～206.5 μg·mL^-1（r=0.9992）范围内浓度与峰面积有良好的线性关系,平均回收率分别为98.0%和97.1%,检出限分别为0.12 μg·mL^-1和0.21 μg·mL^-1,定量限分别为0.36 μg·mL^-1和0.52 μg·mL^-1。结论 本方法准确度高,重复性好,检测灵敏度符合检测要求,可用于化妆品原料2-羟乙基脲及其杂质尿素的含量测定方法。}     

HPLC同时测定盐酸萘甲唑啉滴鼻液中9种抑菌剂

目的 建立HPLC同时测定盐酸萘甲唑啉滴鼻液中4类9种常见抑菌剂硫柳汞、羟苯乙酯、苯甲酸钠、羟苯甲酯、羟苯丙酯、羟苯丁酯、山梨酸钾、苯扎氯铵和苯扎溴铵的含量。方法 采用Ultimate XB-C₁₈（4.6 nm×250 nm,5 μm）色谱柱;以乙腈（A）-0.1 mol·L^-1磷酸二氢钠（磷酸调pH 3.7）为流动相,梯度洗脱,流速1.0 mL·min^-1,柱温35℃,检测波长222 nm,样品温度5℃。结果 9种抑菌剂可完全分离,平均回收率为98.2%~100.6%,RSD为0.5%~1.5%。30批样品中检出硫柳汞、羟苯乙酯、苯扎溴铵和苯甲酸钠4种抑菌剂。结论 该法可用于盐酸萘甲唑啉滴鼻液中抑菌剂的质量控制。     

HPLC法测定大青龙汤颗粒剂中盐酸麻黄碱和盐酸伪麻黄碱的含量

摘 要 目的:建立HPLC测定大青龙汤颗粒剂中盐酸麻黄碱和盐酸伪麻黄碱含量的方法,为该制剂的质量控制提供依据。方法: 采用Agilent Zorbax SB-C₁₈色谱柱(150 mm×4.6 mm,5 μm),以乙腈-含0.3%三乙胺的0.02 mol·L^-1磷酸二氢钾溶液（用磷酸调节至pH=3.0）(5∶95)为流动相,流速:1.0 ml·min^-1,波长：207 nm。结果:盐酸麻黄碱在64～640 μg·ml^-1浓度范围内线性关系良好（r=0.999 6）,平均回收率为101.66%,RSD为2.59%(n=6);盐酸伪麻黄碱在31.8～318 μg·ml^-1浓度范围内呈现良好的线性关系（r=0.999 7）,平均回收率为99.70%,RSD为1.58%(n=6)。结论:该法简便易行,结果准确,可用于大青龙汤颗粒剂的质量控制。     

RP-HPLC法测定注射用羟喜树碱脂质体含量

目的 建立测定注射用羟喜树碱脂质体含量的方法。方法 采用反相高效液相色谱法。色谱系统为ODS柱,以甲醇-水(60∶40)为流动相,检测波长为266 nm。结果 本品在5.886～10.930μg·ml^-1范围内呈线性关系,回归方程为:A=6.438×10⁴C-2.184×10⁴,r=0.9998。回收率为98.9%,RSD%为0.38(n=9)。结论 本法可测定注射用羟喜树碱脂质体中羟喜树碱的含量,方法快速、简便、准确。     

依诺沙星滴眼液中防腐剂含量的测定

目的建立依诺沙星滴眼液中常用防腐剂苯扎溴铵、苯扎氯铵和羟苯乙酯的HPLC含量测定方法。方法采用C18柱,苯扎溴铵和苯扎氯铵以0.005mol/L的醋酸铵溶液(每1L中含三乙胺10mL,用冰醋酸调节pH值至5.0±0.5)-乙腈(40∶60)为流动相,检测波长214nm,进样量为50μL;羟苯乙酯以0.005mol/L的醋酸铵溶液(每1L中含三乙胺10mL,用冰醋酸调节pH值至5.0±0.5)-乙腈(50∶50)为流动相,检测波长256nm,进样量20μL。结果苯扎溴铵、苯扎氯铵和羟苯乙酯在各自线性范围内呈良好的线性关系,r分别为0.9998、0.9998、0.9999;平均回收率分别为100.6%、100.0%、100.2%(n=9)。结论该方法简单、准确,适用于依诺沙星滴眼液中三种常用防腐剂苯扎溴铵、苯扎氯铵和羟苯乙酯的含量测定。     

差示分光光度法测定苯扎溴铵消毒液中主药的含量

目的　建立苯扎溴铵消毒液中苯扎溴铵和亚硝酸钠含量的测定方法。方法　取浓度相同的两份苯扎溴铵消毒液 ,一份加入氨基磺酸铵和 1mol·L-1盐酸作为参比液 ,在 35 4 6nm测定亚硝酸钠的ΔA。取参比液 ,在波长2 6 2 2nm直接测定苯扎溴铵的含量。结果　亚硝酸钠在 0 4～ 2 5 0mg·ml-1浓度范围内 ,线性良好。RSD =0 3%。苯扎溴铵在 0 1～ 3 5 0mg·ml-1浓度范围内 ,线性良好 ,RSD为 1 0 %。结论　该法操作简便 ,灵敏 ,结果满意。     

苯扎溴铵选择性电极的研制

苯扎溴铵原名新洁尔灭属季铵盐类药物,是一种阳离子表面活性杀菌、消毒剂。我国药典采用萃取—容量滴定法测定,操作较麻烦,终点不易判明。近期英、美药典收载的为与苯扎溴铵有相似结构和性质的苯扎氯铵(benzalkonium chloride),他们采用碘酸钾滴定法测定含量,亦有操作麻烦、终点不易判明的问题。本文研制了能直接响应苯扎溴铵离     

苯扎氯铵对大鼠结肠神经及平滑肌的影响

目的研究苯扎氯铵对大鼠结肠神经及平滑肌的影响。方法8~9周龄SD大鼠50只,随机分为两组。氯胺酮麻醉下开腹,实验组用0.1%苯扎氯铵处理大鼠降结肠浆膜40min,温盐水冲洗后关腹,对照组用生理盐水代替苯扎氯铵。分别于术后1、2、3、4、8周取处理段结肠行组织学检查,反转录-聚合酶链反应(RT-PCR)检测乙酰胆碱酯酶(AchE)、巢蛋白(nestin)、乙酰胆碱M3受体(Chrm3)的表达,循环水浴药理学检测平滑肌收缩功能变化。结果苯扎氯铵处理后1周,大鼠结肠神经节细胞明显减少、空泡变,3周后完全消失;结肠平滑肌对乙酰胆碱敏感性增强。结论采用0.1%苯扎氯铵可使大鼠结肠产生去神经作用,结肠平滑肌对乙酰胆碱敏感性增强。     

苯扎溴铵选择性电极的研制

The PVC membrane selective electrode for benzalkonium bromide was studied. It is based on the use of benzalkonium tetraphenylborate ion pair complex as the active material. The PVC membrane electrode showed Nernstian response over the concentration range from 10^-3 mol/L to 5×10^-6mol/L with a slope of 53～57 mV/pC, and the detection limit was found to be 3×10^-6 mol/L. The electrode showed rapid response and good reproducibility. The selectivity coefficients of 16 substances were determined. Using direct potentiometric determination, the recovery of pure benzalkonium bromide was 101.8%. The present method is more rapid and simpler than the volumetric method given in the Chinese Pharmacopoeia.     

Cell viability score (CVS) as a good indicator of critical concentration of benzalkonium chloride for toxicity in cultured ocular surface cell lines

Cytotoxicity of benzalkonium chloride (BAK) is a major factor affecting drug cytotoxicity. This study aimed to determine the critical concentration of BAK for cultured ocular cells, using SIRC (rabbit corneal epithelium), BCE C/D-1b (bovine corneal epithelial cells), RC-1 (rabbit corneal epithelium), and Chang (human conjunctival cells). Cell viability was determined following the exposure of cells to 11 concentrations of BAK for 10, 30, or 60 min using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and neutral red assays, and the cell viability score (CVS) was used to evaluate comprehensively the toxicity of BAK. The CVS system consists of two values. The CVS50 was determined by the number of measurements for viability ⩾50% of control. The CVS40/80 was calculated as follows: CVS40/80 = (number of measurements for viability values >80%) − (number of measurements for viability values <40%). Both %CVS50 and %CVS40/80 decreased with concentrations of BAK. When BAK concentrations were 0.01% or higher, %CVS50 and %CVS40/80 became 0 and less than −90, respectively. Meanwhile, when BAK concentrations were 0.001% or lower, %CVS50 became 100. In the case of %CVS40/80, when the BAK concentrations were 0.002% or lower, the values reached 75 or more, and when 0.0005% or lower, the %CVS40/80 value reached 100. Accordingly, BAK induced very low cytotoxicity in the cultured ocular cell lines at concentrations of 0.002% or lower. The concentration-dependency confirmed that the CVS score is useful for expressing drug cytotoxicity in a simple and comprehensive manner.     

紫外分光光度法测定苯扎溴铵滴眼液的含量

目的：建立苯扎溴铵滴眼液中苯扎溴铵的含量测定方法。方法采用紫外分光光度法测定含量，以262．0nm处为测定波长测定吸收度。结果在0．1～0．6mg／ml之间呈良好线性关系，相关系数v=0．9998，平均回收率为99．7％，RSD为0．45％。结论该法简便、准确，重现性好，可用于苯扎溴铵滴眼液的定量测定。     

Comparison of the ocular surface changes following the use of two different prostaglandin F2α analogues containing benzalkonium chloride or polyquad in rabbit eyes

Objective: The aim of this study is to compare the effect of prostaglandin analogues preserved with either 0.015% or 0.001% benzalkoium chloride (BAK); or 0.001% polyquad (PQ) on the ocular surface of rabbit eyes.

Methods: Forty white rabbits were randomized to receive four-times daily instillation of either 0.0015% tafluprost (TF) preserved with 0.001% BAK (TF-BAK); 0.004% travoprost (TR) with 0.015% BAK (TR-BAK) or 0.001% PQ (TR-PQ); or preservative-free artificial tears in one eye for a 4-week period. Tear samples collected from the 40 rabbits were analyzed by enzyme-linked immunosorbent assays (ELISA) to identify the presence of inflammatory cytokines: interleukin (IL)-1β and IL-6 on day 14. Subsequently, harvested cornea and bulbar conjunctiva were evaluated using light and transmission electron microscopy (TEM).

Results: IL-6 was significantly increased in TF-BAK and TR-BAK groups compared to controls and TR-PQ group (p?=?0.005); however, IL-1β level was not significantly different among four groups (p?=?0.360). Rabbits treated with TR-BAK showed decreased goblet cell density of bulbar conjunctiva and increased pyknotic change and vacuolization of corneal epithelial cells on light microscopy; similar change occurred but was less severe in TF-BAK group. The TR-PQ group showed similar results as the controls. The destruction of the microvillar architecture of bulbar conjunctiva and cornea was most prominent in the TR-BAK group.

Conclusions: Preservatives included in the anti-glaucoma eye-drops showed different ocular surface changes according to the concentration and type in the rabbits. Prostaglandin analogues preserved with higher level of BAK may cause more harmful effects on the ocular surface than PQ-preserved medications.     

Incompatibility of indometacin and benzalkonium in eye drops due to ion-pair formation

Eye drop formulations containing indometacin and benzalkonium salt often show an opalescence as a result of ion-pair formation. The insoluble ion pair was isolated and characterized by infrared and ultraviolet spectroscopy and thin layer chromatography. This ion pair is also responsible for the bad assay results of benzalkonium using the ion-pair extraction method with bromothymol blue as counterion. The distribution     

苯扎溴铵溶液防治皮肤创伤愈合瘢痕的临床观察

目的:观察0.1%苯扎溴铵溶液对皮肤创伤愈合瘢痕的防治疗效。方法:将974例创伤患者随机分为治疗组及对照组各487例,用0.5%的络合碘溶液消毒皮肤,治疗组用3%过氧化氢溶液、0.9%氯化钠注射液、0.1%苯扎溴铵溶液依次清创伤口,缝合后用0.1%苯扎溴铵溶液湿敷包扎伤口,每2 d一次,至伤口愈合,对照组用3%过氧化氢溶液和0.9%氯化钠注射液清创伤口,缝合后常规包扎,观察两组远期皮肤愈合瘢痕的形成。结果:皮肤创伤愈合12月后两组的温哥华瘢痕量表评分差异显著,治疗组低于对照组的评分。结论:0.1%苯扎溴铵溶液对皮肤创伤愈合后的瘢痕形成有防治疗效。