Sensitive determination of itraconazole and its active metabolite in human plasma by column-switching high-performance liquid chromatography with ultraviolet detection

A simple and sensitive column-switching high-performance liquid chromatographic method for the simultaneous determination of itraconazole (ITZ) and its active metabolite, hydroxyitraconazole (HIT) in human plasma is described. ITZ, HIT, and an internal standard, R051012, were extracted from 1 mL of alkalinized plasma sample using n-heptane-chloroform (60:40, vol/vol). The extract was injected onto column I (TSK precolumn BSA-ODS/S, 5 microm, 10 x 4.6 mm ID) for clean-up and column II (Develosil C8-5 column, 5 microm, 150 x 4.6 mm ID) for separation. The mobile phase consisted of phosphate buffer-acetonitrile (68:32 vol/vol, pH 6.0) for clean-up and phosphate buffer-acetonitrile (35:65 vol/vol, pH 6.0) for separation. The peaks were monitored with an ultraviolet detector set at a wavelength of 263 nm, and total time for chromatographic separation was about 24 minutes. The validated concentration ranges of this method were 3 to 500 ng/mL for ITZ and 3 to 1000 ng/mL for HIT. Mean recoveries were 59.7% for ITZ and 72.8% for HIT. Intraday and interday coefficients of variation were less than 4.6% and 5.0% for ITZ, and 4.6% and 4.9% for HIT at the different concentrations. The limit of quantification was 3 ng/mL for both ITZ and HIT. This method was suitable for therapeutic drug monitoring of ITZ and HIT, and was applied to pharmacokinetic studies in human volunteers.     

反相高效液相色谱法测定人血浆中二甲双胍质量浓度

目的建立测定人血浆中二甲双胍质量浓度的方法方法反相高效液相色谱（RP—HPLC）法。色谱柱为EclipseXDB—C8柱（150mm×4．6mm，5μm），以乙腈-甲醇-0．02mol／LKH2PO4。缓冲液（10：34：56，KH2PO4。缓冲液用磷酸调节pH值至3．4）为流动相，流速为1．0mL／min，柱温为室温，检测波长233nm。结果二甲双胍与内标（西咪替丁）的保留时间分别为5．14min和6．46min，二甲双胍检测质量浓度线性范围为17．68～1767．58ng／mL（r=0．9999），方法平均回收率大于94．51％，日内和日间精密度RSD〈8．22％，最低定量限为17．68ng／mL。结论RP—HPLC法操作简便，结果准确、灵敏，适用于二甲双胍血药浓度的测定和药代动力学研究。     

高效液相色谱-荧光分析法测定血浆中替米沙坦浓度

目的建立高效液相色谱-荧光分析（HPLC—FLD）测定人血浆中替米沙坦质量浓度的方法。方法色谱柱为Agilent HC C18柱（250mm×4．6mm，5μm），柱温30％，流动相为磷酸二氢钾缓冲液（磷酸调节pH值为3．7）-甲醇（29：71），流速1．0mL／min，荧光激发波长为300nm，发射波长为366nm。结果替米沙坦和萘普生（内标）的保留时间分别为5．7min和4．4min；替米沙坦质量浓度范围是2．42．484ng／mL（r=0．9997），最低定量质量浓度为2．42ng／mL。结论该法操作简便、选择性好、灵敏度高，可用于替米沙坦的血药浓度测定和药代动力学研究。     

高效液相色谱法测定抗心绞痛新药雷诺嗪的含量

目的建立测定雷诺嗪含量的高效液相色谱法。方法色谱柱为十八烷基硅烷键合硅胶柱(250 mm×4.6 mm,5μm),流动相为乙腈-加0.1 mL三乙胺的0.05 mol/L磷酸二氢钾溶液(50∶50),流速1.0 mL/min,进样量10μL。结果雷诺嗪质量浓度在0.049 2～0.492 g/L范围内与峰面积线性关系良好(r=0.999 3)。结论所用方法简便、快速、准确、专属性强,能有效控制雷诺嗪的含量。     

Determination of donepezil hydrochloride in human plasma and pharmaceutical formulations by HPLC with fluorescence detection

A sensitive, isocratic reversed-phase high performance liquid chromatographic method involving fluorescence detection was developed for the determination of donepezil hydrochloride in tablets and in human plasma. Pindolol was used as an internal standard. Good chromatographic separation was achieved by using an analytical column C18. The system operated at room temperature using a mobile phase consisting of methanol, phosphate buffer (0.02 mol L?1) and triethyl amine (pH 3.5) (55 : 45 : 0.5, V/V/V) at a flow rate 0.9 mL?1 min. The analyte and internal standard were extracted from human plasma via liquid-liquid extraction. The proposed method was validated for sensitivity, selectivity, linearity, accuracy and precision. The calibration curve was linear over the range of 5-2000 ng mL?1 of donepezil with detection limit of 1.5 ng mL?1. Intra- and inter-day relative standard deviations were less than 2.5%. The method was found to be suitable for quality control of donepezil hydrochloride in bulk drug as well as in human plasma.     

阿奇霉素有关物质测定方法的优化分析

目的采用色谱分析法对阿奇霉素相关物质的进行测定,探讨测定方法的优化。方法采用相色谱系统仪器、岛津Class-10Avp系统完成测定,色谱柱为SHISEIDO CAPCELL PAK C18 MG,规格为5μm,250mm×4.6mm。流动相为磷盐酸缓冲液,以0.05mol/L的磷酸氢二钾溶液,以浓度为20%的磷酸调节,直至pH值为8.2;色谱纯试剂乙腈为45∶55;色谱柱温度为30℃;流速为每分钟10mL;检测波长为210nm;进样量为50μL。结果溶液浓度为10~1000μg/mL的范围内的线性关系,结果表明线性关系良好。方法的最低检出限为100ng,r=0.9999。结论本次研究所采用的测定方法能够有效测定阿奇霉素中的相关物质,具有良好的分离性与浓度峰面线性关系。     

在体微透析结合HPLC测定小鼠纹状体细胞外尿嘧啶含量

目的利用在体微透析结合高效液相色谱法建立检测小鼠纹状体细胞外尿嘧啶含量的方法。方法色谱柱为反向VertiSep GES-C18(150 mm×4.6 mm,5μm),流动相为乙腈-0.02 mol/L磷酸二氢钠(2:98),调pH值为3～4,流速为0.8 mL/min,检测波长为260 nm。柱温:0～5℃。结果尿嘧啶的保留时间约为4.5 min,且在0.05～1.0μg/mL范围内线性关系良好(R2=0.998 4)。结论该方法准确、灵敏、方便,重复性好,可用于脑微透析样品中尿嘧啶的含量测定。     

Simultaneous high-performance liquid chromatographic determination of olanzapine and lamotrigine in plasma of bipolar patients

An original method based on the use of high-performance liquid chromatography with both coulometric and diode array detection has been developed for the therapeutic drug monitoring of patients with bipolar disorders being treated with olanzapine and lamotrigine. Chromatographic separation was achieved on a reversed-phase C8 column (150 x 4.6 mm internal diameter, 5 microm) using a mobile phase composed of methanol (27%) and a 50.0 mmol/L, pH 3.5 phosphate buffer (73%). For the analysis of olanzapine and its main metabolite, N-desmethylolanzapine, a coulometric detector was used, with electrode 1 set at -200 mV and electrode 2 at +500 mV. Lamotrigine was determined using a diode array detection at 220 nm. The two detectors were connected in series. For the analysis of biological samples, a clean-up procedure was implemented by means of solid-phase extraction using phenyl cartridges and eluting the analytes with methanol; only a small volume of plasma (150 microL) was needed to analyze both olanzapine and lamotrigine. Linear responses were obtained between 0.1 and 50.0 ng mL(-1) for olanzapine, 0.1 and 25.0 ng mL(-1) for N-desmethylolanzapine, and between 0.25 and 10.0 microg mL(-1) for lamotrigine. The extraction yield values were always higher than 90% for all the analytes, with precision (expressed as relative standard deviation values) lower than 3.4%. The method was applied successfully to some human plasma samples drawn from bipolar patients undergoing combined therapy with the two drugs. Satisfactory values for accuracy were obtained, with mean recovery higher than 91%. Thus, the method appears suitable for the investigation of the chemical-clinical correlations in patients receiving therapy with olanzapine and lamotrigine.     

清脑宁颗粒质量标准研究

目的建立清脑宁颗粒的质量控制方法。方法采用薄层色谱（TLC）法对清脑宁颗粒中赤芍、当归、川芎、陈皮进行定性鉴别;采用高效液相色谱（HPLC）法测定清脑宁颗粒中芍药苷含量,色谱柱为岛津C18柱（250mm×4.6mm,5μm）,流动相为甲醇-0.05mol/L磷酸二氢钾（30∶70）,流速为1.0mL/min,检测波长为230nm。结果TLC法能鉴别出赤芍、当归、川芎、陈皮;芍药苷质量浓度在12～120μg/mL范围内与峰面积呈良好的线性关系,r^2=0.9998（n=6）。结论建立的方法简便可行,专属性强,可用于清脑宁颗粒的质量控制。     

Reversed-phase high performance liquid chromatographic determination of nifedipine in human plasma

A reversed-phase high performance liquid chromatographic method is presented by which the calcium channel blocker, nifedipine (NFP), may be measured in human serum using 17-alpha-ethinylestradiol as an internal standard. A mobile phase of phosphate buffer (0.01 M, pH 6.1)/methanol/acetonitrile (20:35:45) passed through a muBondapak C-18 column at 1.0 ml/min produced symmetrical bands for nifedipine and internal standard. A rapid and simple chloroform extraction of NFP from 1 ml serum samples proved to be efficient and reproducible. Recovery over a concentration range of 5-100 ng/ml was 92.3 +/- 5.1% (mean +/- SD, n = 6). Ultraviolet absorbance detection at 235 nm was sensitive to serum NFP concentrations of 5 ng/ml. Between-day coefficients of variation at 100 and 5 ng/ml were 4.0 and 11.4%, respectively (n = 10). Serum concentration data from NFP-treated subjects are presented.     

RP-HPLC内标法测定大鼠血浆中盐酸氟桂利嗪的含量

目的建立RP-HPLC内标法测定大鼠血浆中盐酸氟桂利嗪含量的方法。方法色谱柱：Shimadzu ODS C18柱（150mm×4.6mm,5μm）;流动相：甲醇-磷酸二氢钾（0.01mol/L,pH=3.5）（68：32）;流速：1.0mL/min;检测波长：254nm;柱温：30℃;进样量：20μL;内标：对羟基苯甲酸丙酯。结果盐酸氟桂利嗪在0.01～0.12μg/mL范围内线性关系良好,r=0.9997（n=7）,信噪比（S/N）〉3时,最低检测浓度为3.48×10-3μg/mL;平均回收率为89.83%,RSD值为2.76%。结论本方法简便、灵敏,回收率和精密度高,可用于盐酸氟桂利嗪新剂型研究中血药浓度的测定及其药代动力学研究。     

高效液相色谱法测定甲硝唑漱口液中甲硝唑含量

目的建立测定甲硝唑漱口液中甲硝唑含量的高效液相色谱法。方法采用Ultimate C18色谱柱(250 mm×4.6 mm,5μm),以甲醇-0.075 mol/L磷酸二氢钾溶液(13∶87)为流动相,流速为1.0 mL/min,检测波长为274 nm。结果甲硝唑质量浓度在50～200μg/mL范围内与峰面积线性关系良好(r=1.000 0,n=5),平均回收率为99.19%,RSD=0.53%(n=6)。结论该方法简便、快速、准确,可用于甲硝唑漱口液的甲硝唑含量测定。     

高效液相色谱法测定盐酸左旋多巴异戊酯的含量

目的建立测定盐酸左旋多巴异戊酯含量的高效液相色谱法。方法色谱柱采用UltimateTMC18柱（250mm×4．6mm5μm）,流动相为0．05mol／L磷酸二氢钾（磷酸调pH=3．5）-乙腈（70：30）,流速为1．0mL／rain,检测波长为280mm,进样量为20μL,柱温为30℃。结果盐酸左旋多巴异戊酯质量浓度在16．1—322．0μg／mL范围内与峰面积线性关系良好（r=0．9999）,平均回收率为98．41％,RSD为1．85％（n=6）。结论该方法准确、简便,精密度高、专属性强,可用于盐酸左旋多巴异戊酯的含量测定。     

RP-HPLC法测定血竭骨刺康胶囊中血竭素的含量

目的建立测定血竭骨刺康胶囊中血竭素含量的方法。方法采用RP-HPLC法。色谱柱为kromasilCls（4．6mm×250mm,5μm）,流动相：乙腈（A）-0．05mol／L磷酸二氢钾（B）,梯度洗脱（0-20min35％-60％A）,流速1．0mL／min,检测波长为440am,柱温：30％。结果血竭素高氯酸盐在0．408-2．040μg内与峰面积呈良好的线性关系,相关系数为0．9998;平均回收率为96．5％,RSD=1．3（n=5）。结论本方法可用于血竭骨刺康胶囊的质量控制。     

An improved sensitive assay for simultaneous determination of plasma haloperidol and reduced haloperidol levels by liquid chromatography using a coulometric detector

A simple, highly sensitive and specific high-performance liquid chromatographic method that uses a coulometric detector for the simultaneous assay of haloperidol and reduced haloperidol in human plasma has been developed, using bromoperidol as the internal standard. A reversed phase C8 5-microns column (25 x 0.46 cm) and a mobile phase of phosphate buffer (pH 7.0), acetonitrile, and methanol (40:20:40 vol/vol) are used for separating the analytes. The analytes are extracted from alkalinized plasma using a mixture of pentane and isopropanol (95:5 vol/vol) and purified by back extraction into a perchloric acid solution. Teflon tubes with screw caps are used throughout the extraction work. The compounds are oxidized at a potential of +0.90 V against a Ag/AgCl reference electrode. The detection limit of the assay is 50 ng/L using 1 mL of plasma. The average interassay coefficient of variation for samples of concentration 1-40 micrograms/L is approximately 8%. Possible drug interferences in the assay have been studied. The absolute recovery of the method is approximately 80%. The assay has been validated by quantitating 150 schizophrenic patients' samples.     

Measurement of duloxetine in blood using high-performance liquid chromatography with spectrophotometric detection and column switching

A method using high-performance liquid chromatography (HPLC) with column switching and ultraviolet (UV) spectroscopy was developed for the determination of duloxetine in human plasma. After centrifugation and addition of venlafaxine as internal standard, plasma samples were injected into the HPLC system and precleaned on a column (10 x 4.0 mm) filled with cyanopropyl (CN)-modified silica of 20 microm particle size, with use of 8% (vol/vol) acetonitrile in deionized water as eluent. Duloxetine was eluted and separated on a LiChrospher 100 CN (5-microm particle size; column size, 250 x 4.6 mm I.D.) using acetonitrile-water-potassium dihydrogenphosphate trihydrate buffer (pH, 6.4; 50:50 vol/vol) and detected at 218 nm. Duloxetine could be analyzed within 30 minutes. The limit of quantification was 5 ng/mL. At duloxetine concentrations up to 138 ng/mL that resulted from therapeutic doses of 30 to 120 mg per day, the interassay reproducibility of quality control samples was better than 12%. The method was found to be robust and stable. With the exception of chlorprothixene and desmethylclomipramine, other drugs that may be used as comedication were not found to exhibit retention times similar to duloxetine. In serum samples from 37 patients treated with 30 to 120 mg per day for at least 7 days, the mean steady state serum concentration of duloxetine was 40 ng/mL, the median was 37 ng/mL, and the 25th and 75th percentiles were 22 and 55 ng/mL, respectively. At 60, 90, and 120 mg/day, the mean +/- SD serum concentrations were 33+/-22.0, 43+/-22.2, and 48+/-17.0 ng/mL, respectively. There was a statistically significant correlation (P<0.05, r2=0.26) between prescribed daily doses and serum concentrations of duloxetine. In patients without or with comedication with other drugs, such as inhibitors of cytochrome P450 2D6 (eg, metoprolol or propranolol), serum concentrations of duloxetine were not significantly different. HPLC with column switching and ultraviolet detection as described here is suitable for pharmacokinetic studies and therapeutic drug monitoring of duloxetine.     

高效液相色谱法测定布洛芬片中布洛芬含量

目的建立高效液相色谱法测定布洛芬片中布洛芬的含量。方法色谱柱为Zorbax—Extend C18柱（150mm×4．6mm，5μm），甲醇-0．01m01／L磷酸二氢钾一磷酸（700：300：0．1）为流动相，流速为1．0mL／min，检测波长为263nm。结果布洛芬质量浓度线性范围是100～800μg／mL，r=0．9999，其低、中、高3个量级的平均回收率分别为99．4％，98．8％，100．2％，RSD分别为0．8％，0．7％，1．0％。结论该法测定布洛芬中布洛芬的含量，结果可靠、简便、准确。     

苍术中非法添加硫酸阿托品及氢溴酸东莨菪碱的检测方法

目的建立中药苍术中非法添加硫酸阿托品及氢溴酸东莨菪碱的定性定量检测方法。方法采用薄层色谱法定性、高效液相色谱法定量;色谱柱为phenomenex Luna C18柱（200 mm×4.6 mm,5μm）,流动相为0.07 mol/L磷酸钠溶液（含0.017 5 mol/L十二烷基硫酸钠,用磷酸调节pH至6.0-乙腈（70∶30）,流速为1.0 mL/min,柱温为30℃,检测波长为216 nm。结果硫酸阿托品及氢溴酸东莨菪碱进样量分别在1.97～9.85μg和2.03～10.15μg的范围内与峰面积积分值线性关系良好,平均加样回收率分别为99.67%和99.77%,RSD为0.12%和0.19%（n=6）。结论该方法简便、快速、准确,可用于检测苍术中非法添加的硫酸阿托品及氢溴酸东莨菪碱。     

复方氯己定达克罗宁乳膏含量测定方法研究

目的建立反相高效液相层析(RP-HPLC)法同时测定复方氯己定达克罗宁乳膏中两种成分含量的方法,用于制定复方氯己定达克罗宁乳膏的质量标准。方法采用C18(250 mm×4.6 mm,5μm)色谱柱,流动相,乙腈∶0.05moL/L磷酸二氢钾溶液(含0.07%庚烷磺酸钠,0.4%三乙胺,用磷酸调pH至3.0),流速1.0 mL/min,检测波长284 nm,柱温25℃。结果盐酸氯己定、盐酸达克罗宁的线性范围分别为0.100 03～2.000 60μg和0.015 18～0.303 60μg,相关系数均为0.999 99,回收率(n=9)分别为99.83%和99.89%,相对标准偏差(RSD)分别为0.41%和0.14%。结论该方法简便、准确,可作为控制复方氯己定达克罗宁乳膏中盐酸达克罗宁、盐酸氯己定含量的质量标准。     

高效液相色谱法测定麻氟滴鼻液中两组分的含量

目的建立测定麻氟滴鼻液中盐酸麻黄碱和地塞米松磷酸钠含量的方法。方法采用高效液相色谱（HPLC）法，色谱柱为Hypersil BDS C18柱（250mm×4．6mm，5μm），流动相为0.025mol／L磷酸二氢钾（加入0．5％三乙胺，用磷酸调节pH值至3．5）-甲醇-乙腈（52：38．4：9．6），流速为0．9mL／min，检测波长为257nm，柱温为40℃。结果盐酸麻黄碱质量浓度在100～600μg／mL范围内与峰面积线性关系良好（r=0．9998），平均回收率为100．54％，RSD=1．0％（n=6）；地塞米松磷酸钠质量浓度在10～60μg／mL范围内与峰面积线性关系良好（r=0．9998），平均回收率为100．20％，RSD=1．2％（n=6）。结论HPLC法专属性好，操作简便、快速，结果准确，可用于麻氟滴鼻液的质量控制。