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OBJECTIVE： To establish UPLC fingerprint of Fortunella margarita， and to conduct its cluster analysis and principal component analysis. METHODS： UPLC method was adopted. The determination was performed on Waters Acquity UPLC BEH C18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution （gradient elution） at the flow rate of 0.3 mL/min. The detection wavelength was set at 330 nm， and sample size was 2 μL. Using fortunellin as reference， UPLC fingerprints of 8 batches of F. margarita were determined. The similarity of 8 batches of samples was evaluated by TCM Chromatographic Fingerprint Similarity Evaluation System（2012 edition） to confirm common peak. Cluster analysis and principal component analysis were performed by using SPSS 24.0 software. RESULTS： There were 24 common peaks in UPLC fingerprints of 8 batches of sample，the similarity of which was higher than 0.97. Cluster analysis showed that 8 batches of samples were clustered into 2 categories. S1， S2， S3， S4， S6， S7 and S8 were clustered into one category； S5 was clustered into the other category. By principal component analysis， the accumulative contribution rate of three main components was 81.366％. CONCLUSIONS： Established UPLC fingerprint， the results of cluster analysis and principal component analysis can provide reference for quality control of F. margarita.
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OBJECTIVE： To optimize and improve the quality standard for Citrus reticulata formula granules. METHODS： Totally 13 batches of C. Reticulata formula granules from 4 different manufacturers were used as trial samples， and qualitative identification of hesperidin and nobiletin in the samples were carried out by TLC according to the method of 2015 edition of Chinese Pharmacopoeia （part Ⅳ）. The quantitative analysis of naringin， hesperidin， hesperetin， nobiletin and tangeretin in C. reticulatae formula granules were conducted by UPLC[The determination was performed on Waters Acquity UPLC BEH C18 column with mobile phase consisted of acetonitrile-0.2％ phosphoric acid aqueous solution （gradient elution）. The detection wavelength was set at 283 nm， and sample size was 3 μL]. RESULTS： The results of TLC showed that in the chromatograms of samples， same color spots were shown in the corresponding positions of the chromatogram of reference substance. The results of UPLC showed， that the linear range of naringin， hesperidin， hesperetin， nobiletin and tangeretin were 0.64-6.44， 15.78-157.80， 0.17-1.66， 2.08-20.85 and 2.04-20.43 μg/mL， respectively （all r≥0.999 2）； the limits of detection were 0.03， 0.33， 0.10， 0.20 and 0.06 μg/mL； the limits of quantitation were 0.07， 1.34， 0.20， 0.60 and 0.22 μg/mL. The average recoveries were 99.4％， 99.6％， 99.7％， 99.7％ and 99.7％ （n＝9）； RSDs of precision （n＝6）， stability （n＝7） and reproducibility （n＝6） tests were all≤2.03％； naringin was detected in only 3 batches of samples from one manufacturer （the content ranged from 0.067 3 to 0.069.6 mg/g）， while the other 4 components were detected in 13 batches of samples （the contents of them ranged 0.646 5-1.728 0， 0.102 6-0.290 5， 0.023 1-0.689 8， 0.018 2-0.270 7 mg/g）. CONCLUSIONS： In this study， the quality standard of C. reticulata formula granules was improved by qualitative and quantitative methods， and the contents of hesperidin， hesperetin， nobiletin and tangeretin were not less than 0.60， 0.10， 0.02 and 0.01 mg/g， respectively.
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ObjectiveTo establish the proper review rules for the microscopic screening of urine samples tested by automatic urinalysis work station which is composed of LabUMat urine dry chemical analyzer and Urised urine sedimental analyzer.Methods The paired comparison was used to analyze the results tested by microscopy and Urised.A total of 2015 random urine samples were enrolled to establish and validate review rules.All the samples were collected from the inpatients and ontpatients of General Hospital of the People's Liberation Army from May to November 2011 and tested by urinalysis work station.2015 urine samples were firstly tested by urinalysis work station,including both urine dry chemical analyzer and urine sediments analyzer.Then each urine sample was examined microscopically by two technicians-in-charge using double-blind method.The average results from the two technicians were used as review results.Compared with review results,we set up the review rules and evaluated the Irue positive rate,false positive rate,true negative rate,false negative rate (omission diagnostic rate) and review rate.According to different test methods by automatic urinalysis work station,four microscopic review protocols were defined:( 1 ) Protocol 1:based on chemistry results only,microscopy review was performed when any of WBC,RBC,PRO and NIT was positive; (2) Protocol 2:based on urine sedimental analysis only,microscopy review was performed when any of WBC,RBC and CAST count was over upper limit of the reference range ; (3) Protocol 3:if any of BLD vs.RBC,LEU vs.WBC was different between two systems,or quantitative results had two or more than two gradient differences,microscopy review was performed; (4) Protocol 4:if any of BLD vs.RBC,LEU vs.WBC was different between two systems,or CAST was over upper limit of the reference range,or alam appeared,microscopic review was performed.300 randomly selected urine samples were tested to validate the review rules.Omission diagnostic rate and review rate were used to evaluate the rules.Results According to our review rules,the positive samples rate was 41.14％ (829/2015) and the negative rate was 58.86％ ( 1186/2015 ) ; Positive samples were composed of RBC (50.30％),WBC (53.32％) and CAST (3.74％).The review rates of four protocols were 42.93％ (865/2015),39.70％ (810/2015),29.58％(596/2015),18.91％ (381/2015 ),respectively.The false negative rates (omission diagnostic rates) were 6.36％ (128/2015),4.42％ (89/2015),1.34％ (27/2015)and 1.04％ (21/2015)respectively.Protocol 4 was selected as an ideal plan.Additional 300 urine samples were tested using protocol 4 in order to confirm the review rule.The review rate,consistency rate,true positive rate,false positive rate,true negative rate,omission diagnostic rate were 19.67％ (59/300),91.67％ (275/300),35.67％ (107/300),7.67％(23/300),56.00％ (168/300),0.67％ (2/300),respectively.After image review revised,the review rate was 8.67％ (26/300).ConclusionThe review rules established by our research for Urinalysis Work Station can find the abnormal urine samples effectively and exactly and can reduce the workload significantly.(Chin J Lab Med,2012,35:810-814)