HPLC波长切换法测定氨咖黄敏胶囊4个成分的含量

摘 要 目的： 建立HPLC波长切换法同时测定氨咖黄敏胶囊中4个成分的含量。方法: 采用Agilent ZORBAX SB C₁₈色谱柱（250 mm×4.6 mm,5 μm）,以乙腈（A）-甲醇（B）-磷酸二氢铵溶液（取0.1 mol·L^-1磷酸二氢铵溶液1 000 ml ,加磷酸1 ml,混匀）（C）为流动相,梯度洗脱,流速1.0 ml·min^-1,柱温 35℃,变换波长时间为（0～9 min ：225 nm;9～38 min ：450 nm）。结果: 采用HPLC波长切换法测定氨咖黄敏胶囊4个成分的含量,线性范围分别为：对乙酰氨基酚24.680～394.900 μg·ml^-1（r=0.999 9）,马来酸氯苯那敏0.201～3.214 μg·ml^-1（r=0.999 9）,咖啡因1.129～18.070 μg·ml^-1（r=0.999 9）,胆红素0.010～0.165 μg·ml^-1（r=0.999 8）;平均回收率分别为：99.25% (RSD=0.46%), 99.29% (RSD=0.32%),99.49% (RSD=0.48%)及99.75% (RSD=0.55%)（n=6）。结论:该法简单,灵敏,准确,重复性好,可用于氨咖黄敏胶囊的含量测定。     

HPLC波长切换法同时测定茵栀祛黄胶囊中7种成分的含量

摘 要 目的：建立HPLC波长切换法同时测定茵栀祛黄胶囊中栀子苷、甘草苷、芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚的含量。方法: 采用Waters Sunfire C18（250 mm×4.6 mm,5 μm）色谱柱;流动相：甲醇（A） 0.05%磷酸溶液（B）（梯度洗脱）,流速为1.0 ml·min^-1 ,柱温为25℃,检测波长(0~15 min：在238 nm波长下检测栀子苷和甘草苷;15~50 min：在254 nm波长下检测芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚),进样量为10 μl。结果: 栀子苷、甘草苷、芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚的线性范围分别为0.13～3.18 μg·ml^-1（r=0.999 8）、0.19～4.84 μg·ml^-1（r=0.999 7）、0.28～7.02 μg·ml^-1（r=0.999 9）、0.13～3.16 μg·ml^-1（r=0.999 9）、0.61～15.27 μg·ml^-1（r=0.999 9）、0.32～8.03 μg·ml^-1（r=0.999 9）、0.39～9.81 μg·ml^-1（r=0.999 9）;平均加样回收率分别为98.84%（RSD=0.74%）、99.34%（RSD=0.86%）、99.54%（RSD=0.30%）、99.56%（RSD=0.80%）、99.85%（RSD=0.41%）、99.57%（RSD=0.70%）、99.64%（RSD=0.30%）（n=9）。结论: 本文建立的HPLC含量测定方法,具有操作简便、专属性高、重复性良好、结果准确可靠的特点,可用于茵栀祛黄胶囊的质量控制。     

HPLC法测定吲哚美辛呋喃唑酮栓中两组分的含量

摘 要 目的：建立同时测定吲哚美辛呋喃唑酮栓中两组分含量的HPLC法。方法: 采用XTerra RP18色谱柱(250 mm×4.6 mm,5 μm),以甲醇为流动相A,0.01 mol·K^-1磷酸二氢钾溶液 三乙胺(100〖KG*9〗∶〖KG-*2〗0.02)为流动相B,梯度洗脱,流速1.0 ml·min^-1,柱温30 ℃,检测波长263 nm,进样量10 μl。结果: 吲哚美辛呋喃唑酮栓中呋喃唑酮和吲哚美辛的分离度符合要求;线性范围分别为5.12~81.87 μg·ml^-1 (r=1.000 0)和3.78~60.45 μg·ml^-1 (r=1.000 0);平均回收率分别为99.6% (RSD=1.5%,n=6)和100.3% (RSD=1.6%,n=6);定量限分别为0.02 μg·ml^-1和0.05 μg·ml^-1。结论:该法经方法学验证,适用于吲哚美辛呋喃唑酮栓的质量评价。     

HPLC法同时测定蒲药灌肠液中3个有效成分的含量

摘 要 目的： 建立蒲药灌肠液中香蒲新苷、异鼠李素-3-O-新橙皮苷和延胡索乙素的HPLC含量测定方法。方法: 采用ZORBAX SB-C₁₈色谱柱（250 mm×4.6 mm,5 μm）,以乙腈-0.1%磷酸（三乙胺调节pH至6.0）为流动相,梯度洗脱程序,流速:1.0 ml·min^-1,检测波长为254 nm（0～14 min）和281 nm（14～25 min）,柱温:30 ℃。结果: 香蒲新苷、异鼠李素-3-O-新橙皮苷和延胡索乙素的线性范围分别为19.840～198.400 μg·ml^-1（r=0.999 6）、20.520～205.200 μg·ml^-1（r=0.999 8）和10.040～100.400 μg·ml^-1（r=0.999 7）,回收率分别为98.8%、98.6%和98.9%,RSD分别为1.4%、1.6%和1.3%（n=6）。结论: 该方法灵敏度高,专属性强,可用于蒲药灌肠液的质量控制。     

化妆品中三种紫外吸收剂检测方法的建立

摘 要 目的：建立高效液相色谱法同时分析化妆品中亚苄基樟脑磺酸、樟脑苯扎铵甲基硫酸、4-甲基苄亚基樟脑这3种紫外吸收剂的方法。方法: 化妆品（水基、乳液类）经甲醇-乙腈-醋酸铵水溶液-四氢呋喃（30∶35∶30∶5,v/v）或者（膏霜、粉质类）经乙腈-甲醇-四氢呋喃（30∶20∶50, v/v）涡旋、超声提取后检测;采用Thermo scientific C18（250 mm×4.6 mm,5 μm）色谱柱,以甲醇、乙腈、0.02 mol·K^-1醋酸铵水溶液（冰醋酸调至pH 5.15）为流动相,梯度洗脱,柱温：35℃,流速：1.0 ml·min^-1 ,DAD 检测器,检测波长288 nm、300 nm。结果: 亚苄基樟脑磺酸、樟脑苯扎铵甲基硫酸、4 甲基苄亚基樟脑分别在12.26～98.06 μg·ml^-1（r=0.999 9）、9.31～74.48 μg·ml^-1（r=0.999 9）、6.86～54.88 μg·ml^-1（r=0.999 9）范围内呈良好的线性关系,检出限及定量限范围分别为4.6,6.4,1.6 ng和17.4,15.9,5.5 ng,平均回收率分别为99.2%（RSD=1.58%）、99.8%（RSD=2.38%）、99.2%（RSD=2.03%）（n=36）。结论:本方法简便快捷,重复性良好,结果准确可靠,适用于化妆品中紫外吸收剂的含量测定。     

GC-MS法同时测定麝香保心丸中6种活性成分的含量

摘 要 目的：建立GC-MS法同时测定麝香保心丸中龙脑、异龙脑、肉桂醛、肉桂酸、麝香酮、苯甲酸苄酯6种活性成分的含量。方法: 采用气相色谱 质谱联用(GC MS)法,色谱柱为DB 5(30 m×0.25 mm, 0.1 μm)毛细管柱;程序升温：70℃恒温2 min,以5℃·min^-1)的速率升温至150℃,维持4 min,以20℃·min^-1)的速率升温至260℃,维持3 min;检测质荷比范围：10～425。结果: 龙脑、异龙脑、麝香酮、苯甲酸苄酯、肉桂醛、肉桂酸分别在0.022～22.000 μg·ml^-1(r=0.999 6),0.024～24.000 μg·ml^-1(r=0.999 6),0.028～28.000 μg·ml^-1 (r=0.999 8),0.034～34.000 μg·ml^-1 (r=0.999 6),0.040～40.000 μg·ml^-1 (r=0.999 7),0.050～50.000 μg·ml^-1 (r=0.999 9)范围内与峰面积呈良好的线性关系。龙脑、异龙脑、肉桂醛、肉桂酸、麝香酮、苯甲酸苄酯的平均加样回收率分别为97.20%,97.40%,97.53%,99.60%,98.78%和98.27%,RSD分别为0.89%,1.18%,1.52%,1.49%,0.79%和1.74%(n=6)。结论:该方法准确、有效、重复性好,可用于麝香保心丸多成分的质量控制研究。     

多波长HPLC法同时测定皮肤康洗液中5种有效成分的含量

摘 要 目的：建立多波长切换高效液相色谱法同时测定皮肤康洗液中芍药苷、甘草苷、甘草酸铵、大黄素和蛇床子素5种有效成分的含量。方法: 色谱柱为 Diamonsil C18柱(200 mm×4.6 mm,5 μm);流动相为乙腈 0.1%盐酸水溶液,梯度洗脱;流速为1.5 ml·min^-1;柱温为30℃;检测波长为232 nm(0～6 min,检测芍药苷)、277 nm(6～10 min,检测甘草苷)、254 nm(10 min以后,检测甘草酸铵、大黄素、蛇床子素）。结果: 芍药苷、甘草苷、甘草酸铵、大黄素和蛇床子素的线性范围分别为28.20～282.0 μg·ml^-1（r=0.999 7）、12.150～121.500 μg·ml^-1（r=0.998 8）、13.420～134.200 μg·ml^-1（r=0.999 5）、0.047～0.466 μg·ml^-1（r=0.999 9）、2.380～23.800 μg·ml^-1（r=0.999 9）,平均加样回收率分别为98.49%,99.00%,98.38%,97.36%,97.70%,RSD分别为0.71%,0.62%,0.85%,0.92%,0.78%（n=6）。结论:该方法简便、准确、灵敏度高、重复性好,可用于同时测定皮肤康洗液中上述5个有效成分的含量。     

HPLC法同时测定大败毒胶囊中绿原酸、咖啡酸和阿魏酸的含量

摘 要 目的: 建立HPLC法同时检测大败毒胶囊中绿原酸、咖啡酸和阿魏酸的测定方法。方法: 采用Agilent C₁₈（250 mm×4.6 mm,5 μm）色谱柱;乙腈-0.4%磷酸溶液（15∶85）为流动相,流速：1.0 ml·min^-1;检测波长：324 nm。柱温:30℃,进样量：10 μl。结果:线性范围：绿原酸5.42～32.43 μg·ml^-1(r=0.999 8),咖啡酸2.63～26.32 μg·ml^-1(r=0.999 4),阿魏酸1.44～14.44 μg·ml^-1(r=0.999 9);平均回收率：绿原酸99.98%,RSD=0.2%(n=6),咖啡酸99.31%,RSD=0.4%(n=6),阿魏酸99.48%,RSD=0.8%(n=6)。结论:该方法操作简便、准确,重复性好,可用于测定大败毒胶囊中绿原酸、咖啡酸和阿魏酸的含量。     

HPLC法测定枳实消痞丸中芸香柚皮苷、柠檬苦素、和厚朴酚及厚朴酚含量

摘 要 目的： 建立高效液相色谱法同时测定枳实消痞丸中芸香柚皮苷、柠檬苦素、和厚朴酚及厚朴酚含量的分析方法。方法: 色谱柱为岛津Shim-pack VP-ODS C18柱（250 mm×4.6 mm,5 μm）,流动相A为乙腈 甲醇(1∶2),流动相B为水,梯度洗脱,流速为1.0 ml·min^-1,柱温为30℃,进样量为20 μl;芸香柚皮苷检测波长为λ1=283 nm,柠檬苦素检测波长为λ2=210 nm,和厚朴酚、厚朴酚检测波长为λ3=294 nm。结果: 芸香柚皮苷、柠檬苦素、和厚朴酚、厚朴酚分别在5.26～105.20μg·ml^-1（r=0.999 8）、7.65～153.00 μg·ml^-1（r=0.999 4）、6.21～124.20 μg·ml^-1（r=0.999 3）、6.45～129.00 μg·ml^-1（r=0.999 6）范围内线性关系良好,平均加样回收率(n=6)分别为99.00％（RSD=0.77％）、98.17％（RSD=1.19％）、98.78％（RSD=0.86％）、97.90％（RSD=0.99%）。结论:该方法简便、准确、可靠,可用于枳实消痞丸的质量控制。     

HPLC法测定黄连上清片中4种成分的含量

摘 要 目的： 提高黄连上清片质量标准,建立同时测定绿原酸、栀子苷、黄芩苷、盐酸小檗碱4种成分含量的方法。方法: 采用高效液相色谱法,色谱柱为DiamonsilTM C₁₈(250 mm×4.6 mm,5 μm),流动相为二元梯度系统,其中溶剂A为乙腈,溶剂B为0.3%磷酸水溶液,流速为1.0 ml·min^-1,检测波长为238 nm,柱温30℃,进样量为10 μl。结果: 绿原酸、栀子苷、黄芩苷、盐酸小檗碱的线性范围分别为8.11～81.10 μg·ml^-1(r=0.999 7)、13.08～130.80 μg·ml^-1(r=0.999 7)、10.76～107.60 μg·ml^-1(r=0.999 8)、7.92～79.20 μg·ml^-1(r=0.999 8)范围内呈良好的线性关系,平均加样回收率分别为99.19%(RSD=0.9%)、98.44%(RSD=1.1%)、99.12%(RSD=1.0%)、99.18%(RSD=1.1%)(n=9)。结论:该方法简便、准确、重复性好,可用于黄连上清片的质量控制。     

氯卡麻滴鼻剂中氯霉素、盐酸麻黄碱和硫酸卡那霉素的含量测定

目的建立氯卡麻滴鼻剂中主药成分氯霉素、盐酸麻黄碱和硫酸卡那霉素的含量测定法,以便更好的控制其质量。方法高效液相色谱法,测氯霉素和盐酸麻黄碱,L-column C18色谱柱(250mm×4.6mm,5μm),流动相为0.02mol/L磷酸二氢钾溶液-甲醇(60:40)为流动相;检测波长254nm。流速1.0ml/min,柱温35℃,进样量为10μl;紫外分光光度法,测硫酸卡那霉素含量,测定波长530nm。结果氯霉素其检测质量浓度在25μg/ml～100μg/ml范围内与峰面积积分值的线性关系良好(r=0.9999),平均回收率为100.46%,RSD=1.2%(n=9);盐酸麻黄碱其检测质量浓度在100μg/ml～400μg/ml范围内与峰面积积分值的线性关系良好(r=0.9992),平均回收率为99.50%,RSD=0.49%(n=9);硫酸卡那霉素其检测质量浓度在0.1mg/ml～0.3mg/ml范围内与紫外吸收度的线性关系良好(r=0.9999),平均回收率为99.50%,RSD=0.97%(n=9)。结论本法简便、准确、重现性好,可用于氯卡麻滴鼻剂中各主成分含量测定。     

Kanamycin induced ototoxicity in the laboratory rat

Two comparative experiments on the behavioural, audiometric and histological effects of kanamycin-induced cochleotoxicity in the Wistar rat are reported.In the first experiment kanamycin 400–1,500 mg/kg/day was injected subcutaneously for 20 days and the morphological damage to the organ of Corti assessed. In the second experiment the progression of damage to the organ of Corti was examined in animals given kanamycin 700 mg/kg/day for 4–20 days. Behavioural audiometric studies of threshold shift were undertaken throughout both experiments.In the first study, all the animals were killed after a recovery period of 20 days from the last injection, i.e., day 40, and in the second study groups of animals were killed at 4-day intervals between days 4 and 20 of dosing.One cochlea from each animal was critical point-dried, dissected to expose the organ of Corti and examined by scanning electron microscopy. A vertical section through the contralateral cochlea was examined by light microscopy. The results of the morphological examination of the cochleas were collated with the behavioural audiometry.The morphological damage to the organ of Corti followed a stereotyped pattern of degeneration, the extent of which appeared to be determined both by the number and concentration of the kanamycin administrations. The collateral audiometric examinations indicated that extensive damage had taken place before a shift in the behavioural auditory threshold could be detected by observation of the Preyer reflex.     

Kanamycin priming for audiogenic seizures in mice

It is well established that mice can be made to suffer epileptic-like seizures when subjected to certain kinds of loud noises. Some strains of mice are genetically susceptible while others can be made susceptible by using acoustic priming techniques. It has been suggested that priming mice with a loud bell sound at a critical development age causes hair cell damage which is a necessary part of the audiogenic seizure syndrome. Since ototoxic drugs also cause such damage, non-susceptible BALB/C mice were given kanamycin sulphate 400 mg/kg daily from age 5 days to age 21 days and then tested for audiogenic seizures at age 28 days. Eighteen of the twenty mice which survived the kanamycin treatment displayed full epileptic-like seizures when subjected to loud sounds at age 28 days. Histological surface preparations of the organ of corti from these mice revealed extensive outer hair cell damage in all turns of the cochlea while the inner hair cells and a few outer hair cells in the apical turn appeared undisturbed. All of the cochlea nerve fibres appeared normal in the histological preparations. The relationship of these results to the phenomenon of recruitment is discussed.     

Kanamycin acetyltransferase gene from kanamycin-producingStreptomyces kanamyceticus IFO 13414

A kanamycin producer,Streptomyces kanamyceticus IFO 13414 is highly resistant to kanamycin. Cloning of the kanamycin resistance genes inS. lividans 1326 with pIJ702 gave several kanamycin resistant transformants. Two transformants,S. lividans SNUS 90041 andS. lividans SNUS 91051 showed similar resistance patterns to various aminoglycoside antibiotics. Gene mapping experiments revealed that plasmids pSJ5030 and pSJ2131 isolated from the transformants have common resistant gene fragments. Subcloning of pSJ5030 gave a 1.8 Kb gene fragment which showed resistance to kanamycin. Cell free extracts ofS. lividans SNUS 90041,S. lividans SNUS 91051 and subclone aS. lividans SNUS 91064 showed kanamycin acetyltransferase activity. The detailed gene map is included.     

比浊法测定硫酸卡那霉素注射液的效价

目的　建立测定硫酸卡那霉素效价的方法。方法　采用比浊法,并与管碟法相比较。结果　比浊法线性范围为2～1 0U·ml-1 ,线性方程:A =1 .1 0 4 0 - 1 .2 6 5 9logC(r=0 .992 5 ) ;平均回收率为97.1 1 %(n =4 )。结论　比浊法与管碟法的测定结果无显著性差异,可作为硫酸卡那霉素的效价测定方法之一,且更快捷、方便。     

染料比色法测定硫酸卡那霉素滴眼液中主药的含量

目的:建立以染料比色法测定硫酸卡那霉素滴眼液中主药含量的方法。方法:依据卡那霉素与溴甲酚绿在pH值为6.0时的磷酸盐缓冲液中反应生成离子对化合物的光谱特征,于432nm波长处测定吸收度并计算含量。结果:卡那霉素检测浓度在18.0～72.0μg/ml范围内与吸收度呈良好的线性关系(r=0.9998),平均回收率为99.3%(RSD=1.20%)。结论:本方法快速、简便,可用于该制剂生产过程中的快速检验。     

旋光法测定硫酸卡那霉素滴眼液的含量

目的：测定硫酸卡那霉素滴眼液的含量。方法：采用旋光法测定。结果：平均回收率99.75%(RSD=0.69,n=4)。结论：旋光法简便,可靠,可作为该制剂的质量控制方法之一。     

硫酸卡那霉素注射液中有关物质测定方法研究

目的采用高效液相色谱-蒸发光散射检测器(HPLC-ELSD)法测定硫酸卡那霉素注射液中有关物质。方法采用ZorbaxSB-C18柱(250 mm×4.6 mm,5μm),以0.2 mol/L三氟醋酸溶液-甲醇(95∶5)为流动相,流速为1.0 mL/min,柱温为室温,用蒸发光散射检测器检测(漂移管温度为80℃,载气压力为1.0 bar)。结果以对照溶液质量浓度的对数值与相应峰面积的对数值计算回归方程,其相关系数(r)均大于0.999,二者线性关系良好。结论高效液相色谱-蒸发光散射检测器法简便、准确、专属、灵敏。     

Viability of Human Melanocytes HEMa-LP Exposed to Amikacin and Kanamycin

Aminoglycosides, such as amikacin and kanamycin, are powerful broad-spectrum antibiotics used for the treatment of many bacterial infections. The widely used aminoglycosides have the unfortunate side effects of targeting sensory hair cells of the inner ear, so that treatment often results in permanent hair cell loss. The aim of the study was to evaluate the influence of incubation time and drug concentration on viability of melanocytes cultured in the presence of amikacin or kanamycin. The normal human melanocytes HEMa-LP and the different concentrations of amikacin (0.075, 0.75 and 7.5 mmol/l) and kanamycin (0.06, 0.6 and 6.0 mmol/l), were used. The estimations were performed after 24, 48 and 72 h. The observed decrease in melanocytes viability may be an explanation for the mechanisms involved in aminoglycosides toxicity on pigmented tissues during high-dose and/or long-term therapy.     

柱前衍生化HPLC检测硫酸卡那霉素的方法研究

目的 采用柱前衍生化高效液相色谱法建立了快速定性、定量分析食品中的硫酸卡那霉素的方法。方法 首先茚三酮-吡啶溶液作为衍生化试剂考察了衍生化反应条件。包括反应溶液的酸碱度、温度和反应时间,并建立了色谱分离条件。衍生化条件为在pH为5．8条件下,75℃水浴中反应20min。色谱条件为,色谱柱：150×4．6mm Kramasd C18;流动相：甲醇-水（40：60）;流速：O．5mL·min^-1;紫外检测波长390nm;进样量：20μL。结果 硫酸卡那霉素的浓度在2．24×10^-5mol·L^-1～1．12×10^-4mol·L^-1之内,浓度与峰面积呈现良好的线性关系,建立的方法线性良好,样品回收率在95％～98~%之间。结论 本方法快速、灵敏,适合日常样品检测的需要。