IntroductionCannabidiol (CBD) has a variety of pharmacological effects including antiepileptic, antispasmodic, anxiolytic and anti-inflammatory among other pharmacological effects. However, since CBD is a terpene-phenolic compound, its clinical application is limited by its poor water solubility, low stability, and low bioavailability.MethodsIn this study, we used several strategies to address the above problems. Hydrochloric acid was used to modify zein to improve the molecular flexibility. Flexible zein nanoparticles (FZP-CBD) loaded with CBD was prepared to improve the stability and bioavailability of CBD. The parameters were evaluated in terms of morphology, particle size (PS), polydispersity index (PDI), zeta potential (ZP), entrapment efficiency (EE%), loading capacity (LC%), and storage stability. Simulated gastrointestinal fluid release experiment and bioavailability assay were applied in the evaluation.ResultsThe simulated gastrointestinal fluid experiment showed that the release rates of FZP-CBD and natural zein nanoparticles (NZP-CBD) loaded with CBD were 3.57% and 89.88%, respectively, after digestion with gastric fluid for 2 h, 92.12% and 92.56%, respectively, after intestinal fluid digestion for 2 h. Compared with NZP-CBD, the Cmax of FZP-CBD at 3 different doses of CBD was increased by 1.7, 1.3 and 1.5 times respectively, and AUC0-t was increased by 1.4, 1.1 and 1.7 times respectively, bioavailability (F) was increased by 135.9%, 114.9%, 169.6% respectively.DiscussionThe experimental results showed that FZP-CBD could protect most of the CBD from being released in the stomach, and then control its release in the intestines, promote the absorption of CBD in the small intestine, and increase the bioavailability of CBD. Therefore, FZP-CBD could improve the utilization value of CBD and provide a new idea for the application of CBD in medicine and pharmacy.
质量控制是中药现代化和国际化的关键.充分认识中药质量控制研究中的关键科学问题对于更好地理解中药至关重要.本文系统讨论了中药质量控制中的关键科学问题如质控指标和替代对照品,为符合中医特色的中药质量控制研究提供指导.
Objective: To establish a GC-FID method to determine the content of 17 phthalic ester plasticizers in adsorbed tetanus vaccine. Methods: The protein was precipitated with acetonitrile, and the layers were separated by salting out with sodium chloride. The supernatant was transferred and analyzed by GC-FID with external standard method. Agilent HP-5（30 m×0.32 mm×0.25 μm） capillary column was adopted. The temperature program: Initial temperature of 50 ℃ was maintained for 1 min, then was increased to 230 ℃ at the rate of 20 ℃·min -1 and was maintained for 3 min. The temperature was increased to 300 ℃ at a rate of 5 ℃·min -1 and maintained for 5 minutes. Column flow rate was 1 mL·min -1 . The temperatures of detector and inlet were 280 ℃ and 250 ℃ respectively. The linear velocity of the carrier gas（nitrogen） was set to 19.148 cm·s -1 . The injection volume was 1 μL and the split mode was splitless. The sampling mode: automatic injection. Results: The 17 plasticizers showed good linear relationship in the range of 0.5-20 μg·mL -1 with a correlation coefficient more than 0.99. At three different concentrations(15, 10, 5 μg·mL -1 ) of spiked samples, the recovery rates of 17 plasticizers ranged from 83.6% to 111.2%, with RSD between 0.39% and 3.9%. No plasticizers were detected in 5 batches of adsorbed tetanus vaccine by this method. Conclusion: The method has good specificity, high accuracy and precision, good solution stability, and high durability. It can be used to determine contents of plasticizers in adsorbed tetanus vaccine drugs.
Objective: To establish an HPLC method for simultaneous determination of three isomers in captopril API and tablets. Methods: A Chiralpak IG chiral column（250 mm×4.6 mm, 5 μm） was adopted. The mobile phase A was n-hexane-trifluoroacetic acid（100∶0.1） and the mobile phase B was anhydrous ethanol-trifluoroacetic acid（100∶0.1） with gradient elution. The flow rate was 1.0 mL·min -1 . The temperature of the column was set at 35 ℃, and the wavelength was set at 215 nm. The injection volume was 10 μL. Results: The method had a good specificity, and no interferences from the excipients were found. Captopril and its three isomers were well separated. The average recoveries were 99.6%, 102.1% and 93.6%, with RSD of 1.7%, 1.4% and 2.0%, respectively. The RSD of repeatability were all less than 2.0%. The test solution was stable after 24 h. The quantification limits of diastereoisomer Ⅰ,enantiomer and impurity F were 0.91,0.99 and 3.23 μg·mL -1 ,respectively. The detection limits were 0.27, 0.30 and 0.97 μg·mL -1 , respectively. The calibration curve showed good linearity over the range of 1.82-45.51 μg·mL -1 of diastereoisomer Ⅰ, 0.99-39.71 μg·mL -1 of enantiomer, 3.23-40.33 μg·mL -1 of impurity F（r was 0.999 0-1.000 0）, respectively. The contents of impurity F in three batches of captopril were 0.04%,0.04% and 0.03%, respectively. The contents of impurity F in three batches of captopril tablets were 0.03%, 0.03% and 0.03%, respectively. No diastereoisomer Ⅰ or enantiomer were detected in captopril and tablets. Conclusion: The methodology validation results showed that the method is suitable for the determination of three isomers in captopril and tablets.
Objective: To establish an HPLC-ELSD method for the simultaneous determination of Polysorbate（Tween） 20 and Polyethylene Glycol（PEG） 4000 in Paliperidone palmitate injection. Methods: The separation was performed on a TSK gel 3000 PWxl（7.8 mm×300 mm） column with 20 mmol·L -1 ammonium acetate aqueous solution and acetonitrile（60∶40） as the mobile phase by isocratic elution. And the flow rate was 0.6 mL·min -1 . The column temperature was 25 ℃ and the sampling volume was 20 μL. The ELSD was maintained at 50 ℃ throughout the determination. The nebulizer gas flow rate was 2.0 L·min -1 （0 ℃,1 atm）. Results: PEG4000 showed good linearity within the range of 0.3-0.75 mg·mL -1 （r=0.999 4）. Tween 20 showed good linearity within the range of 0.12-0.3 mg·mL -1 （r=0.999 6）. The minimum detection limit of PEG4000 and Tween 20 were 0.02 mg·mL -1 ,0.05 mg·mL -1 , respectively. The minimum quantitative limit of PEG4000 and Tween 20 were 0.03 mg·mL -1 , 0.07 mg·mL -1 , respectively. The repeatability RSD of PEG4000 and Tween 20 were 2.3% and 1.9%, respectively. The average recoveries of PEG4000 was 98.7% and the RSD was 0.90%（n=6）. The average recovery of Tween 20 was 96.3% and the RSD was 0.80%（n=6）. Conclusion: The method for simultaneous determination of Tween 20 and PEG4000 in Paliperidone palmitate injection by using HPLC-ELSD is precise and suitable.
Objective: To establish an HPLC method and chemical pattern recognition model of poeny leaves from Henan province, providing quality evaluation of poeny leaves. Methods:Analysis was performed on an Agilent C 18 column（250 mm×4.6 mm, 5 μm） with acetonitrile-0.1% formic acid for gradient elution. The flow rate was 1.0 mL·min -1 , the detection wavelength was set at 270 nm, and the column temperature was 35 ℃. “Similarity Evaluation System of Fingerprint of Tradition Chinese Medicine（2012 edition）” was adopted to establish reference fingerprint and to calculate the similarities of 21 batches of poeny leaves. Combining with principal component analysis and cluster analysis, the overall quality of poeny leaves was evaluated. Results:There were 18 common characteristic peaks in the fingerprints of the 21 batches of samples, 5 peaks were identified as gallic acid, ethylgallate, paeoniflorin, ellagic acid and luteoloside. Similarities of 21 batches of samples were between 0.907-0.979. Samples were clustered into 3 types by principal component analysis which consistented with cluster analysis result. Conclusion:The method combining HPLC fingerprint with chemical pattern recognition is simple and reliable, which can quickly evaluate the quality and consistency of different resources of poeny leaves from Henan province, and is suitable for the evaluation of the overall quality evaluation of poeny leaves.
Objective: To establish PCR identification technology of Poria cocos（Schw.） Wolf, a Chinese medicinal material of the fungus, and identify the authenticity of Poria cocos（Schw.） Wolf from the perspective of molecular biology. Methods: The classic CTAB method was improved and used to extract the genomic DNA from the samples, specific primers were designed with the internal transcription spacer of Poria cocos（Schw.） Wolf as the target sequence, and PCR reaction system was optimized. PCR identification method of Poria cocos（Schw.） Wolf was established. At the same time, the target band specifically amplified was cloned and sequenced to identify its accuracy. Finally, the method was verification. Results: The extracted genomic DNA was of high concentration, good purity, and good integrity. The results of agarose gel electrophoresis showed that the genuine Poria cocos（Schw.） Wolf could appear a single bright specific amplified band at 199 bp, but the counterfeit could not appear amplified band. The specificity of the primer was 100%. It was cloned and the target fragment was clear and bright after the electrophoresis pattern. And the cloned fragment had 100% similarity with sequence of Poria cocos（Schw.） Wolf registered in Genbank database. When the operator didn’t know the authenticity of the samples, the samples were tested, and the position of the target bands were consistent with the expectation. The sensitivity test results showed that the lower limit of DNA detection was up to 1 ng·μL -1 . Three laboratory staff were selected for testing the authenticity of Poria cocos（Schw.） Wolf, and the results were the same and consistent with expectations. The established PCR identification method showed good specificity, high accuracy and repeatability. Conclusion: PCR identification technology provides a simple, reliable, specific and highly accurate identification method for the authenticity of Poria cocos（Schw.） Wolf. Its standard reference materials can be successfully stored by using molecular cloning technology and the sequencing results are accurate and reliable.
Based on the published scientific literatures, the research progress and application of reciprocating holder apparatus are described. Reciprocating holder apparatus with flexible and specific holders can meet the requirements of research and development for transdermal delivery system, extended-release preparation, ocular implants, intravaginal rings, drug eluting stents and other special dosage forms or combination products. Reciprocating holder apparatus has a broad application prospect and may provide scientific support for the in vitro-in vivo correlation research.
Dissolution test is an important way to evaluate the quality of oral solid dosage forms. The dissolution and absorption of drugs in gastrointestinal tract are continuous and dynamic progress. Due to the lack of chemical components and fluid dynamics conditions which are similar to in vivo enviroment, the traditional dissolution methods are extremely limited in the evaluation of drug in vivo. Therefore, in vitro non-biological dynamic dissolution methods based on physiological state of gastrointestinal tract have been constructed, including multi-compartment dissolution models, dissolution-absorption systems, and in vitro digestion models. These novel in vitro dissolution models could be supplements or substitutes of traditional dissolution models, so that researchers can comprehensively understand the drug dissolution behavior in vivo. This paper reviewed recent advances of non-compendial dissolution models, and summarized the applicable scopes and relative merits, aiming to provide a reference for the evaluation of dissolution degree of oral drugs.
Objective: To establish and validate an ELISA method to measure the titer of botulinum antitoxin type A. Methods: Botulinum toxoid type A was used as coated antigen coated enzyme label plate, and diluted serum samples were added after plate blocking. The horseradish peroxidase labeled rabbit anti-horse IgG（IgG-HRP） was subsequently used as the enzyme-labeled antibody. Tetramethyl benzidine（TMB） was added for 10-15 min for color development, and the reaction was terminated by 2 mol·L -1 of sulfuric acid. Finally, the A 450 nm value was read on the enzyme marker. The logarithm of concentration（X） of the sample concentration of standard curve was taken as the horizontal coordinate and the A 450 nm value（Y） was taken as the vertical coordinate to draw the standard curve. The serum sample concentration could be obtained by substituting A 450 nm value of serum samples to be tested into the curve equation, and the titer of the serum samples could be obtained by multiplying the dilution ratio. Results: Under the optimized conditions, the specificity of the method was high, there was no cross reaction with B, C, D, E and F type of antitoxin serum except botulinum antitoxin type A. In the linear range of 50-400 mIU·mL -1 , the linear relationship was high with r＞0.990 0. The accuracy was between 80% and 120% and the RSD of precision（repeatability and intermediate precision） was less than 20%. The titers for 20 batches of botulinum antitoxin type A serum were determined by ELISA in the range of 525-2 450 IU·mL -1 , and the neutralization test was in the range of 675-2 400 IU·mL -1 . The coefficient of correlation r=0.906 0（P＜0.001）, which was significantly positive correlated between the two methods. Conclusion: The established ELISA method could acquire stable results under laboratory conditions, which provides a simple detection method for botulinum antitoxin type A in vitro.
Objective: To develop a SPE-isotope dilution GC-MS/MS method for the determination of the residues of 18 polycyclic aromatic hydrocarbons（PAHs） in root and rhizome herbs. Methods: Using isotope as internal standard, the sample was extracted by acetonitrile and purified by solid phase extraction on C 18 cartridges. GC-MS/MS method was used for the assay. The chromatographic column was DB-17ms（30 m ×0.25 mm, 0.25 μm） with temperature programming and MRM detection. Results: The calibration curves for the 18 kinds of typical PAHs were linear in the range of 2-50 ng·mL -1 . The average recovery rate was in the range of 63.8%-118.0%, with RSD 0.45%-19.0%（n=3）. The LOQs were 0.5-1 μg·kg -1 . Conclusion: This method is specific, sensitive, and can be used for residue detection of 18 typical kinds of PAHs in root and rhizome herbs.
Objective: To establish an ultra-high performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS） method for the simultaneous determination of twenty chemical components（arecoline hydrobromide, adenosine cyclophosphate, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, liquiritin, benzoylmesaconine, benzoylaconitine, isoliquiritin, benzoylhypacoitine, liquiritigenin, isoliquiritigenin, glycyrrhizic acid, 6-gingerol, honokiol, costunolide, dehydrocostus lactone, magnolol, betulinic acid）in Shipi powder. Methods: The separation was performed over 40.0 min on a Shim-pack GIST C 18 column（100 mm×2.1 mm, 2.0 μm） with a mobile phase consisting of 0.1% formic acid in water and acetonitrile with gradient elution（0-12 min, 2%B→14%B;12-24 min, 14%B→51%B;24-26 min, 51%B→70%B;26-36 min, 70%B;36-37 min, 70%B→2%B;37-40 min, 2%B） at a flow rate of 0.3 mL·min -1 . The column temperature was 35 ℃ and the injection volume was 1 μL. The detection was carried out by electrospray ionization（ESI）. Positive and negative electrospray ionizations were performed in multiple reaction monitoring（MRM） mode. Results: All of the analytes showed good linearity（r≥0.997 6） in the tested ranges. The precision, repeatability and stability of the method were good for the twenty chemical components. The average recoveries were in the range of 93.4%-108.7% with relative standard deviations RSD≤5.0%. The contents of arecoline, adenosine cyclophosphate, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, liquiritin, benzoylmesaconine, benzoylaconitine, isoliquiritin, benzoylhypacoitine, liquiritigenin, isoliquiritigenin, glycyrrhizic acid, 6-gingerol, honokiol, costunolide, dehydrocostus lactone, magnolol, betulinic acid in 15 bantches of in Shipi powder were 7 190.26-13 726.74, 90.11-301.93, 9.65-71.13, 102.88-710.98, 40.51-179.73, 10.90-71.22, 3 424.43-27 337.63, 508.34-1 276.14, 108.27-192.65, 95.74-378.92, 565.19-1 018.53, 106.23-623.42, 3.53-16.47, 6 104.50-15 933.93, 1 047.58-1 750.15, 537.91-2 582.74, 332.83-581.28, 348.22-760.81, 350.65-2 189.73, 2.06-39.22 μg·g -1 . Conclusion: The established method is accurate, stable and reproducible, and can provide a research method for the quality control of Shipi powder.
Objective: To investigate the difference of contents of extract, astilbin and engeletin among different cross sections color of Smilacis Glabrae Rhizoma, and to analyze the correlation between color and component content. Methods: 26 Batches of Smilacis Glabrae Rhizoma with different cross sections were collected, and their color was observed by naked eye and numbered according to light reddish brown and off-white cross sections respectively. The color value of Smilacis Glabrae Rhizoma powder was determined by color difference meter, its moisture was determined by oven drying method, the content of extract was determined by alcohol hot soaking method, and the contents of asxinobin and flavarin were determined by HPLC. Cluster analysis, partial least square method and correlation analysis were used to sort out and analyze the data. Results: The color of the chromatic meter measuring value well represented the actual medicinal material color. Partial least squares analysis results showed that the color value a* was the main parameters affecting the classification. Correlation analysis showed that the contents of extract, astilbin and engeletin were significantly positively correlated with color values a * and b * （P＜0.01）, and significantly negatively correlated with color values L * and Eab * （P＜0.01）. Conclusion: There is a very significant correlation between the content of the extract astirin and engeletin of Smilacis Glabrae Rhizoma, and the color of different sections. The difference in the content of components between the two section colors is mainly reflected in the higher content of the light red-brown section than the light white section, which provides a new way for the quality of Smilacis Glabrae Rhizoma.
Objective: To establish an UPLC method for simultaneous determination of 5-hydroxymethyl furfuraldehyde（5-HMF）, cimicifugoside, calycosin-7-glucoside, cimicifugin, 4’-O-beta-glucopyranosyl-5-O-methylvisamminol, calycosin, sec-O-glucosylhamaudol, 6-gingerol, schisandrin and fraxinellone in Guben Kangmin decoction, so as to analyze the variation of the 10 ingredients after compatibility. Methods: The samples were separated on an ACQUITY UPLC BEH C 18 column（2.1 mm×100 mm, 1.7 μm） by gradient elution with acetonitrile-0.1% formic acid aqueous solution at a flow rate of 0.25 mL·min -1 . The detection wavelength was 254 nm. The column temperature was controlled at 30 ℃. Results:5-HMF,cimicifugoside, calycosin-7-glucoside, cimicifugin, 4’-O-beta-glucopyranosyl-5-O-methyl-visamminol, calycosin, sec-O-glucosylhamaudol, 6-gingerol, schisandrin and fraxinellone exhibited good linearity（r~2=0.999 5-1.000）among the ranges of 156.00-10000, 12.50-1600, 3.12-400, 12.50-1600, 6.25-800, 0.78-100, 3.90-500, 20.50-1600, 0.58-75 and 12.50-200 ng, respectively. The average recoveries（RSD＜3%, n=6） were between 98.3%-102.3%. After the compatibility, the contents of 6-gingerol, calycosin-7-glucoside, 5-HMF, schisandrin and fraxinellone increased significantly（P＜0.05 or P＜0.01）, while there was no significant change in the contents of cimicifugoside, cimicifugin, 4’-O-beta-glucopyranosyl-5-O-methyl-visamminol, calycosin and sec-O-glucosylhamaudol. Conclusion: The method can be used for the quality control of Guben Kangmin decoction and its preparations, and can provide scientific basis for the clinical compatibility and application of Guben Kangmin decoction.
Objective: To study the metabolites and pharmacokinetics of three main pharmacodynamic components of Xiaoer Fengye Kechuanping mixture in rats. Methods: UHPLC-ESI TOF MS and ACQUITY UPLC? HSS T3（100 mm×2.1 mm, 1.8 μm） column with a 0.1% formic acid water and acetonitrile as mobile phase were used. Gradient elution was applied and the flow rate was 0.3 mL·min -1 . ESI with positive and negative ions pattern was used. After intragastric administration, serum, bile, urine, and feces of rats were collected for analyzing. The metabolites were identified by analyzing the mass-to-charge ratios, retention times, and fragments of ions. Meanwhile, UHPLC-MS/MS was used in pharmacokinetic study. Gliclazide were used as internal standard. UPLC? BEH C 18 （50 mm×2.1 mm, 1.7 μm） column was used. The mobile phase of 0.1 % formic acid water and acetonitrile was gradient eluted and the plasma samples were precipitated with methanol. The ESI and MRM were used for quantification. The selected monitoring ions were ephedrine m/z 166.2→148.1, glycyrrhetinic acid m/z 471.2→453.2, p-coumaric acid m/z 162.9→119.1, and internal standard m/z 322→170. Xiaoer Fengye Kechuanping mixture was gavaged by 2, 5 and 10 mL·kg -1 , respectively. DAS 3.1 software was used to calculate the pharmacokinetic parameters after the plasma concentrations of three components were determined. Results: 3 prototypes and 10 metabolites which have Ⅰ phase（demethylation, oxidation） metabolism and Ⅱ phase（methylation, esterification and glucuronic acid sulfate） metabolism were identified. Meanwhile, the specificity of the three components in plasma met the requirements without interference, and all linear relationships were good（r＞0.995 0）, the extraction recovery and matrix effect were within the requested range. The stability of different conditions met the requirements. All three components fit the two-compartment model. Conclusion: The three main efficacy components of Xiaoer Fengye Kechuanping mixture are identified by the UHPLC-ESI TOF MS. The p-coumaric acid mainly metabolizeds in Ⅱ phases. Meanwhile, the plasma concentration of three components in rats are analyzed by UHPLC-MS/MS to calculate pharmacokinetic parameters. It is found that the absorption rate of glycyrrhetinic acid is slower than that of other two components. This paper studied the processes of Xiaoer Fengye Kechuanping mixture in the body and provide theoretical basis for exploring the pharmacodynamic material basis.
EGFR-TKIs（epidermal growth factor receptor-tyrosine kinase inhibitor） has been widely used in lung cancer targeted therapy. Monitoring drug concentration has extensive significance for guiding clinical medication. Through the database search for the methods of determining lung cancer targeted drugs concentration in blood, urine, and cerebrospinal fluid, current domestic and foreign biological sample drug detection methods are summarized and analyzed.Present domestic and foreign lung cancer targeted drug measurement saremainly blood drug concentration monitoring. Drug concentration measurementsin the urine or cerebrospinal fluid are rare.HPLC-MS/MS method is mainly used in drug concentration monitoring and the analysis conditions are different. Besides, related research in China is insufficient. This paper can provide ideas for researchers before developing quantitative methods and selecting detectors.
Objective: To verify the slope ratio method for determination of anticoagulant activity of leeches in laboratory and to investigate. The feasibility of slope ratio method for quality control of leech anticoagulant activity. Methods: Seven batches of leeches and Japanese medical leeches were used to prepare the extracts with different concentrations in parallel. The anticoagulant activity was determined by slope ratio method after the activated partial thromboplastin time（APTT） was measured. Specificity, linearity range, precision, accuracy and durability were systematically investigated in a laboratory. Results: The slope ratio method was used to measure the anticoagulant activity of leeches. The activity of leech extract decreased after heating at 100 ℃. The experiment was not established after adding low molecular weight heparin. The anticoagulant activity of Japanese medical leeches was determined by leeches as the control material, which indicated that the method had better specificity and selectivity. Accuracy inspection: the relative deviations of the 5 relative level values of 64%, 80%, 100%, 125% and 156% were all less than ±20%, and the logarithm of the theoretical value of potency（horizontal coordinate） and the logarithm of the corresponding value of potency（vertical coordinate） showed an obvious linear relationship, with the correlation coefficient greater than 0.95. The slope is between 0.95 and 1.05, indicating good accuracy of the method. The geometric coefficient of variation（GCV, %） of each relative potency level was less than 20%. RSDs of many batches of decoction pieces were all less than 20%, indicating that the precision of this method was good. Linearity and range test: the method could be used to accurately determine 64% to 156% relative to the control medicinal materials, and the linear range meets the requirements. Durability test: leech extract could be placed for 24 h and heated at 37 ℃ for 1 h without any change in activity, indicating that the durability of this method was good. Conclusion: The slope ratio method is precise, accurate and reproducible in the determination of anticoagulant activity of leeches, which is suitable for the quality control of leeches.
Objective: To research the molecular structure and formation route of the degrade products of bacteriostatic agents in compound clotrimazole cream, and to provide evidence for improvement of preparation formulation. Methods: This work, which included the determination of bacteriostatic agents and their degrade products and the compatibility study between paraben bacteriostatic agent and excipient potassium hydroxide, was carried out by using HPLC to measure the content of bacteriostatic agents and their degrade products and by using UPLC-HRMS to characterize the structure of the degrade impurity. HPLC was applied with a Phenomenex Prodigy ODS3 C 18 column（150 mm×4.6 mm, 5 μm）, and gradient elution with 0.02 mol·L -1 ammonium acetate solution（A）-methanol（B） under the condition of detection wavelength 257 nm and column temperature 35 ℃. UPLC-Q-Orbitrap HRMS was applied with a YMC C 18 column（250 mm×4.6 mm, 5 μm）, and gradient elution with 0.1% formic acid in water（A）-0.1% formic acid in acetonitrile（B） under the MS condition of HESI source and negative ion mode. Results: Through compatibility test simulating cream process conditions and HRMS analysis, it was clarified that excipient potassium hydroxide promoted the more serious degradation of paraben antibacterial agents, and the incompatibility of the above two components was the cause of multiple batches of antibacterial agents that did not meet the requirement（＜80% of the prescription amount）. Conclusion: The selection of potassium hydroxide as an excipient in the prescription of compound clotrimazole cream by individual company has the risk of unqualified antibacterial content. It is recommended that the related company optimizes the cream prescriptions and strives to reduce the safety risks caused by the excipient-excipient incompatibility.
Objective: To develop a method for simultaneous determination of the plasma drug concentrations of tegacyclin and amikacin in infection in intensive care unit（ICU） patients with carbapenem resistant enterobacteriaceae（CRE） using LC-MS/MS, and apply it to the therapeutic drug monitoring（TDM）. Methods: NaOH solution and methanol solution with internal standards of minocycline and kanamycin were added into plasma samples. The resulting solution was then extracted with the mixed solution of ethyl acetate and methylene chloride. The supernatant was dried with nitrogen and re-dissolved in methanol. Determination was carried out with a waters Cortec C 18 （100 mm×4.6 mm, 2.7 μm） chromatographic column. Mobile phase A was methanol, mobile phase B was formic acid/ammonium formate aqueous solution, the flow rate was 0.3 mL·min -1 , and the injection volume was 2 μL. Electrospray spray ion source, positive ion scanning and multiple reaction monitoring mode were used. Results: The linear range and quantitative limit of tegacyclin and amikacin both met the requirements. The inter-and intra-day precision and matrix effect were all ＜15%. Samples of ten patients treated with tegacyclin（50 mg, q12h） and amikacin（0.8 g, qd） were collected. The steady-state trough concentrations of tegacyclin and amikacin in ten ICU patients with CRE infection were（83.86±84.56） ng·mL -1 and（83.86±84.56） ng·mL -1 , respectively. There were large individual differences caused by the two antibiotics. Conclusion: This method is rapid, sensitive, specific and reproducible. It can be used for clinical TDM of ICU patients with CRE infection.
Objective:To establish a method for simultaneous determination of 37 kinds of hair dyes such as p-phenylenediamine in oxidized hair dye products with quantitative analysis of multi-components by single marker（QAMS）, and verify its applicability and accuracy in the quality control of hair dye products. Methods:The analytes were separated by high performance liquid chromatography（HPLC）. TechMate AQUA-STⅢ（250 mm×4.6 mm, 5 μm） column was used and a mixture of the ammonium acetate solution containing 4% acetonitrile and acetonitrile was used as mobile phase for gradient elution. The determination was carried out at 235 nm and 280nm by diode array detector. m-aminophenol was used as the internal standard to determine the relative correction factors(f si ) of the 36 other hair dyes. Contents of the dyes in sample were calculated by their f si . Results: Good linearity with correlation coefficients greater than 0. 999 was obtained for all the analytes. Recoveries of the method were from 81.7% to 114.8% at three concentration levels, with the reproducibility RSD were less than 5.0%（n=6）. The contents of hair dyes in 6 batches of oxidative hair dye products measuredby QAMS method were not significantly different from those using the standard curve method（SCM）. The relative average deviation（RAD） were less than 4%. Conclusion:The established QAMS method with simple operation, accurate results and saving reference materials, which can quantitatively analyze 37 kinds of hair dyes in oxidized hair dye products at the same time. It can provide reference for quality evaluation and risk monitoring of oxidized hair dye products.
