LC-MS/MS法测定人血浆中复方磺胺甲噁唑的浓度及其临床应用

摘要：目的:建立LC-MS/MS法同时测定人血浆中磺胺甲噁唑（SMZ）和甲氧苄啶（TMP）浓度的方法。方法:以甲苯磺丁脲为内标,血浆样品经含内标的甲醇溶液沉淀后,采用LC-MS/MS法测定,流动相A为水（含0.1%甲酸）,流动相B为乙腈（含0.1%甲酸）,梯度洗脱,采用电喷雾离子阱正离子模式,以多离子反应监测的方式进行正离子扫描,监测SMZ m/z 254.093→156.000,TMP m/z 291.144→230.000,内标m/z 271.242→154.900。结果:血浆中SMZ在2～500μg·ml-1（r=0.996 3）,TMP在0.2～50μg·ml-1（r=0.993 0）范围内线性关系良好;SMZ和TMP的日内和日间RSD≤10.55%,SMZ和TMP方法准确度分别为86.33%～97.38%,89.08%～97.17%;样本在室温放置8 h、4℃放置24 h、冰冻放置7 d和反复冻融3次等条件下均稳定,RSD≤8.91%。临床样本监测结果显示,SMZ峰浓度均值为（119.14±53.9）4μg·ml-1,TMP峰浓度均值为（7.49±3.60）μg·ml-1。结论:本方法可快速、准确、灵敏监测人血浆中复方磺胺甲噁唑的浓度,可用于临床样本中复方磺胺甲噁唑血药浓度的测定。     

系数倍率法同时测定复方磺胺甲噁唑片中磺胺甲噁唑和甲氧苄啶的含量

目的建立复方磺胺甲噁唑片中磺胺甲噁唑（SMZ）和甲氧苄啶（TMP）的含量测定方法。方法采用系数倍率法,分别在235nm和257nm处测定吸光度,从而测定复方磺胺甲噁唑片中SMZ和TMP的含量。结果 SMZ的线性范围为6～16μg.mL-1（r=0.9994）,平均回收率为98.07%,RSD为0.65%（n=6）;TMP的线性范围为1.2～3.2μg.mL-1（r=0.9987）,平均回收率为98.91%,RSD为0.66%（n=6）。结论该法简便易行、重现性好、结果可靠,可用于该制剂的质量控制。     

RP-HPLC同时测定小儿复方磺胺甲噁唑片中两组分的含量

目的建立同时测定小儿复方磺胺甲噁唑片中磺胺甲噁唑(SMZ)和甲氧苄啶(TMP)的含量的反相高效液相色谱法.方法采用Kromasil C18色谱柱,以0.025 mol/L磷酸溶液(用20%氢氧化钠调节pH 3.0±0.5)-乙腈(75∶25)为流动相,流速:1.0 ml/min,检测波长:240 nm.结果 SMZ在20～200 μg/ml范围内,峰面积与其浓度线性关系良好(r=0.9997),加样回收率为100.1%;TMP在 4～40 μg/ml范围内,峰面积与其浓度线性关系良好(r=0.9999),加样回收率为96.0%.结论本法操作简便、专属性强.     

RP-HPLC同时测定小儿复方磺胺甲噁唑片中两组分的含量

目的　建立同时测定小儿复方磺胺甲噁唑片中磺胺甲噁唑(SMZ)和甲氧苄啶(TMP)的含量的反相高效液相色谱法。方法　采用KromasilC18色谱柱,以0. 025mol/L磷酸溶液(用20%氢氧化钠调节pH 3. 0±0. 5) 乙腈(75∶25)为流动相,流速: 1. 0ml/min,检测波长: 240nm。结果　SMZ在20~200μg/ml范围内,峰面积与其浓度线性关系良好(r=0. 9997),加样回收率为100. 1%;TMP在4~40μg/ml范围内,峰面积与其浓度线性关系良好(r=0. 9999),加样回收率为96. 0%。结论　本法操作简便、专属性强。     

HPLC法测定复方磺胺甲噁唑片中SMZ及TMP的含量

目的建立同时测定SMZ和TMP含量的HPLC法.方法 ODS柱,以0.03 mol@L1H3PO4(含0.2%三乙胺)-甲醇(80:20)为流动相,流速为1.2 ml@min,以咖啡因为内标,检测波长为240 nm.结果测得平均回收率SMZ为99.7%,RSD=0.56%;TMP为98.3%,RSD=0.42%.结论本法简便,重现性好,结果准确,可作为该片剂质量控制的方法.     

高效液相色谱法测定复方磺胺甲噁唑混悬剂的含量

目的　采用HPLC法测定复方磺胺甲唑混悬剂中磺胺甲唑 (SMZ)和甲氧苄啶 (TMP)的含量。方法　采用Zor baxSB C18色谱柱 ;以磷酸盐缓冲液 乙腈 (75∶2 5)为流动相 ;流量为 1 0ml/min ;检测波长为 2 3 0nm ;用外标法测定。结果　平均回收率 :SMZ为 99 68% ,RSD为 0 2 6% ;TMP为 10 0 6% ,RSD为 0 76%。结论　该方法简便、准确、可靠 ,适用于复方磺胺甲唑混悬剂的定量检测。     

HPLC同时测定复方磺胺甲噁唑片的两个主药含量

目的建立复方磺胺甲噁唑片中磺胺甲噁唑(sohamethoxazole,SMZ)和甲氧苄啶(trimethoprim,TMP)的HPLC测定方法。方法采用高效掖相色谱法,色谱柱:waters C18(4μm,3.9mm×150mm)柱,流动相为乙腈-磷酸缓冲液(pH6.8±0.1),(12∶88),检测波长为240nm。结果在优化的色谱条件下,SMZ和TMP完全分离,片剂辅料不干扰测定,SMZ在20～100μg/mL范围内呈良好线性关系(r=0.9999),TMP在4～20μg/mL范围内呈良好线性关系(r=0.9999)。ASD<1.5%。结论该法结果准确、简便、快速、专属性强,重复性好,敏感度高,适用于复方磺胺甲噁唑片的含量测定。     

双波长分光光度法测定复方磺胺甲异噁唑混悬剂两主药的含量

采用双波长分光光度法测定复方磺胺甲基异噁唑混悬剂中两主药的含量,相关系数0.9999,平均回收率和精密度分别为SMZ 99.6%,RSD=0.38%;TMP 98.3%,RSD=0.22%.本法操作简便、快速、结果准确、精密度高.     

HPLC法测定复方磺胺甲噁唑颗粒剂的含量

目的:采用HPLC法同时测定复方磺胺甲噁唑颗粒剂中磺胺甲噁唑(SMZ)和甲氧苄啶(TMP)的含量。方法:色谱柱为ODS C_(18)柱,4.6x150mm,流动相为甲醇-磷酸盐缓冲液(pH5.90)(20:80),检测波长为240nm。结果:线性范围分别为:SMZ 20～181μg·ml~(-1),(r=0.9999);TMP 4～38μg·ml~(-1),(r=0.9999)。平均回收率分别为:SMZ100.5％(RSD=0.29％);TMP 100.3％(RSD=0.51％)。结论:本法分离度好,快速,简便,可同时测定该品中的两种组分。     

复方磺胺甲噁唑片的胶束薄层色谱分析

以十二烷基硫酸钠 (SDS)水溶液为展开剂、聚酰胺薄膜为固定相 ,对复方磺胺甲唑片(复方新诺明 )中两组分进行定量分析 .磺胺甲基异唑 (SMZ)和甲氧苄氨嘧啶 (TMP)分别在4 0～ 12 0mmo/L和 3 5～ 10 5mmol/L范围内吸收峰面积和浓度呈良好的线性关系 .相关系数分别为 0 9995和 0 9998,回收率分别为 10 1 4%和 99 5 % ,RSD(% )分别为 0 5 8和 1 6 0 .     

双波长倍增差示法测定复方新诺明片含量

分别测定标准溶液和样品溶液在265及275 nm处的吸收度,根据标准溶液的吸收系数可同时计算出样品中 SMZ 和 TMP 的含量。SMZ 平均回收率与变异系数分别为100.8%,0.95%;TMP 为99.16%,0.82%。方法简便、快速。结果令人满意。     

北京地区2350株急性腹泻病原菌药物敏感性测定

应用WHO推荐的K-B法对1988～1990年分离的2350株急性腹泻病原菌做了9种抗菌药物敏感性测定,并对不同年度的志贺氏菌和沙门氏菌的敏感率及志贺氏菌属不同菌群敏感率进行了分析。 结果表明,志贺氏菌属,沙门氏菌属、弧菌属和病原性大肠杆菌对诺氟沙星和依诺沙星的敏感率分别为97％和96.8％;志贺氏菌属对吡哌酸和庆大霉素的敏感率分别为96.1％和85.6％。沙门氏菌对头孢噻肟的敏感率为100％。弧菌属对新诺明、复方新诺明和氯霉素的敏感率为85.4％、86％和85.7％,而志贺氏菌、沙门氏菌、气单孢菌属相病原性大肠杆菌对新诺明、复方新诺明和氯霉素敏感率很低,只有19.3％、28.3％和49.4%。沙门氏菌属、弧菌属和气单孢菌属对氨苄青霉素的敏感率也很低,分别为43％、32.6％和24.4％。 志贺氏菌属对氯霉素和庆大霉素的敏感率及沙门氏菌属对新诺明、复方新诺明、氨苄青霉素,庆大霉素和诺氟沙星的敏感率年度间差异都非常显著(P<0.005)。志贺氏菌属的四个菌群对吡哌酸、诺氟沙星和依诺沙星的敏感率均无明显差异,而对新诺明和复方新诺明敏感率差异非常显著(P<0.005),其中A、C群敏感率高于B、D群,D群最低。对氯霉素、氨苄青霉素和庆大霉素的敏感率在群间差别也非常显著(P<0.005),D群最高,B群较低。     

HPLC法测定增效联磺制剂中三组分的含量

目的：建立反相高效液相色谱法测定增效联磺制剂中磺胺甲噁唑、磺胺嘧啶与甲氧苄啶三组分的含量。方法：采用Spherisob ODS色谱柱,0．05mol／L磷酸二氢钠-甲醇(75：25)为流动,230nm为检测波长。结果：方法平均回收率：磺胺甲噁唑100．1％,RSD=0．20％(n=6);磺胺嘧啶100．9％,RSD=0．42％(n=6);甲氧苄啶99．6％,RSD0．69％(n=6)。结论：方法简便、快捷、准确,是增效联磺制剂质量控制的理想方法。     

Reproductive Toxicity of Sulfamethazine in Swiss CD-1 Mice during Continuous Breeding

Sulfamethazine (SMZ) was evaluated for reproductive toxicityin Swiss CD-1 mice using a continuous breeding protocol. SMZwas administered in the diet at 0, 0.25, 0.5, or 1% (w/w), whichrepresented an average daily intake of 0, 313, 625, or 1250mg SMZ/kg/day, respectively. Exposure of F0 male and femalemice to 1% SMZ for 126 days resulted in a significant decreasein the mean number of live pups per litter and the number oflitters produced (task 2); the percentage pups born alive to1% SMZ females showed a nonsignificant decrease versus controlfemales. The effects on fertility were rapid to onset (1 to4 weeks) and cumulative in nature. F0 male and female body weightswere slightly depressed from 3 weeks to the end of the study.The crossover mating trial (task 3) revealed that the adverseeffect on ferility involved both treated partners in that littersize decreased when either 1% SMZ males were bred to controlfemales or 1% SMZ females were mated with control males. Afterapproximately 155 days of exposure of F0 mice to 1% SMZ, theterminal body weight of 1% SMZ females was significantly decreasedand that of 1% SMZ males showed a nonsignificant decrease. Inaddition, the liver weight to body weight ratio of the maleswas increased. Further, the prostate and seminal vesicle weightto body weight ratios were decreased in 1% SMZ males relativeto control males. No treatment-related gross or histopathologicallesions were noted for the pituitary or reproductive organsof either sex. Sperm assessment indicated no significant differencein the epididymal sperm concentration or percentage motile orabnormal sperm. In conclusion, SMZ was found to be a reproductivetoxicant in the male and female Swiss CD-1 mouse, albeit atrelatively high dietary intake (1250 mg/kg/day), and in thepresence of mild systemic toxicity.     

正交投影分光光度法测定泻痢停片中TMP和SMZ的含量

目的：建立泻痢停片中TMP和SMZ的含量测定方法。方法：通过正交投影算法消除混合物光谱中干扰组分的光谱,直接测定泻痢停片中的TMP和SMZ的含量。结果：TMP和SMZ的平均回收率分别为99．91％和99．63oA,RSD分别为0．14％和0.61％,样品测定结果与标准方法比较,经t检验（n=6,P〉0．05）表明二者无显著性差异。结论：正交投影分光光度法测定泻痢停片中TMP和SMZ的含量,简便、准确、实用。     

非相关成分参比一倍率导数光谱法测定制剂中甲氧苄啶和磺胺甲(口恶)唑

提出了非相关成分参比一倍率导数光谱法用于测定混合组分体系的基本原理和实验方法。本法可用于待测成分和非相关成分导数光谱严重重叠且待测成分含量较低时的不经分离直接测定。试用本法消除了甲氧苄啶(TMP)和磺胺甲嚼唑(SMZ)及附加剂之间的相互干扰,从而测定了部分制剂中的TMP:用“零交法”测定了SMZ,平均回收率分别为:102.5±1.63%(CV)和100.3±0.99%(CV)。     

Trimethoprim/sulfamethoxazole does not affect the steady-state disposition of indinavir.

This study evaluates the safety and potential pharmacokinetic interaction between indinavir and trimethoprim/sulfamethoxazole (TMP/SMZ). In a randomized, three-period crossover fashion, 12 healthy adults received 1 week of indinavir sulfate 400 mg orally every 6 hours with placebo, TMP 160 mg/SMZ 800 mg orally every 12 hours with placebo, and indinavir sulfate with TMP/SMZ. Plasma indinavir, SMZ, and TMP concentrations were determined after the last dose of each treatment period. Concomitant administration resulted in a 17% decrease in geometric mean trough plasma indinavir concentrations (p = 0.032), an 18% increase in geometric mean AUC0-12 h and Cmax TMP values (p = 0.031 and 0.030, respectively), and a 5% increase in geometric mean AUC0-12 h SMZ values (p = 0.039). None of these effects was considered clinically significant. The combination of indinavir sulfate and TMP/SMZ is generally well tolerated, with no clinically significant pharmacokinetic interaction being noted.     

HPLC法测定复方磺胺甲噁唑分散片两种成分的含量

目的建立测定复方磺胺甲噁唑分散片两种成分的高效液相色谱方法。方法采用HPLC法,kromasil C18柱（4.6×150mm,5μm）;以水-乙腈-三乙胺（800∶200∶1）（用醋酸调pH至5.9）为流动相;检测波长：240nm;流速：1.0mL.min-1;柱温：30℃。结果磺胺甲噁唑进样量在60～600μg.mL-1范围内呈良好的线性,r=0.9999;平均回收率为97.97%,RSD=0.88%;甲氧苄啶进样量在16～160μg.mL-1范围内呈良好的线性,r=0.9999;平均回收率为97.48%,RSD=0.75%。结论本方法操作简便,结果准确可靠,可用于复方磺胺甲噁唑分散片的质量控制。     

Induction of acetylating capacity with complete Freund's adjuvant and hydrocortisone in the rabbit

This study was undertaken to investigate whether modifications in the reticuloendothelial system (RES) activity induce changes in the extent of sulfamethazine (SMZ) acetylation. Five days prior to any RES stimulation or inhibition, pharmacokinetic parameters of SMZ and N-acetyl-SMZ were determined in albino rabbits following a SMZ dose of 20 mg/kg iv. Blood and urine samples were collected and analyzed spectrophotometrically. During the actual test period, one group of rabbits served as control, receiving 0.75 ml of a 0.9% NaCl solution per kg ip on day 0 and then one to seven doses of SMZ over the next 43 days. The effect of RES stimulation was assessed in a second group of rabbits receiving 0.75 ml of complete Freund's adjuvant (CFA) per kg on day 0 followed by seven iv injections of 20 mg of SMZ per kg over the next 43 days. Three groups of rabbits received known RES depressants: colloidal carbon (CC), 32 mg/kg iv; hydrocortisone (HC), 150 mg/kg iv for 10 days; cyclophosphamide (CTX), 15-75 mg/kg iv for 12 days. The administration of each of these compounds was accompanied or followed by one to seven in injections of 20 mg of SMZ per kg. The SMZ metabolic rate constant (km) was not affected in the control, CC, or CTX groups. CFA increased the SMZ km by 60 and 135% in fast and slow acetylators, respectively (p less than 0.05), and HC increased SMZ km by 39% (p less than 0.05). It is concluded that the acetylating capacity of rabbits may be induced by CFA and high doses of HC. Consequently, drug acetylation and RES activity appear to be independent.     

Effects of Ball-Milling and Cryomilling on Sulfamerazine Polymorphs: A Quantitative Study

The effects of ball-milling and cryomilling on sulfamerazine forms I and II (SMZ FI, FII) were investigated using X-ray powder diffraction, infrared and near-infrared (NIR) spectroscopy. Cryomilling resulted in a complete amorphization of both polymorphs. Milling at room temperature gave mixtures of amorphous SMZ (FA) and FII. Calibration models were developed for the quantitative analysis of binary (FI/FII, FI/FA, and FII/FA) and ternary (FI/FII/FA) mixtures using NIR spectroscopy combined with partial least-squares (PLS) regression. The PLS models for binary (0%–100%), ternary (0%–100%), and low-level (0%–10%) binary mixtures had root-mean-square errors of prediction of ≤1.8%, ≤5.1%, and ≤0.80%, respectively. The calibration models were used to obtain a detailed quantitative picture of solid-state transformations during milling and any subsequent recrystallizations. FA prepared by cryomilling FI for less than 60 min recrystallized to mixtures of FI and FII, whereas samples milled for more than 60 min crystallized to pure FII. The effect of comilling SMZ with stoichiometric amounts of additives was investigated. SMZ formed amorphous materials with oxalic, dl-tartaric, and citric acids that were more stable toward recrystallization than FA. Amorphous SMZ/oxalic acid was found to recrystallize to a 2:1 cocrystal during storage. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 103:1766–1778, 2014