Qualitative and quantitative analysis of the major components in Qinghao Biejia decoction by UPLC-Orbitrap Fusion-MS/MS and UPLC-QQQ-MS/MS and evaluation of their antibacterial activities.

OBJECTIVE: In the present study, the chemical components of Qinghao Biejia decoction (QBD) were qualitatively and quantitatively analyzed using UPLC-Orbitrap Fusion-MS/MS and UPLC-QQQ-MS/MS techniques, followed by identification of each component's origin and evaluation of the antibacterial activity of QBD and its components. METHODS: High-resolution mass spectrometry was used to obtain information on the precise molecular weight, retention time, and fragmentation ion peaks of the compounds used to identify the components of QBD and establish a method for their quantification. In vitro assays including determination of the minimal inhibitory concentration and growth curves were used to assess the antibacterial activity of QBD and its components. RESULTS: A total of 39 components, including fatty acids, phenolic acids, amino acids, flavonoids, coumarins, terpenoids, and alkaloids, were identified by UPLC-Orbitrap Fusion-MS/MS. A high-performance analytical method was also established to quantify 12 components of QBD. The content of mangiferin was relatively high (estimated to be 814 µg/g). The results of the antibacterial assays indicated that mangiferin exhibits antibacterial effects against two strains causing respiratory tract infections. CONCLUSIONS: The present study suggests that mangiferin may serve as a natural compound which shows high antibacterial activity. The results can aid the discovery and analysis of the active antimicrobial components present in QBD and further provide a reference for quality assessment of multi-component herbal prescriptions.

Detection of Exogenous γ-Hydroxybutyric Acid in Rat Blood Exosomes.

OBJECTIVES: To find a method to distinguish exogenous gamma-hydroxybutyrate (GHB) from endogenous GHB by establishing ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) based on exosome for quantitative detection of GHB in the rat blood. METHODS: Adult male SD rats were divided into 1 h, 5 h, 10 h administration group and control group. After 1 h, 5 h and 10 h of single precursor of GHB gamma-butyrolactone (GBL) intraperitoneal injection in administration groups, 5 mL blood was collected from the abdominal aorta. Meanwhile, the control group was given a same dose of normal saline, and 5 mL blood was collected at 1 h. Among the 5 mL blood, 0.5 mL was directly detected by HPLC-MS after pretreatment, and exosomes were extracted from the remaining blood by differential centrifugation and detected. RESULTS: The concentration of GHB in the control group was (87.36±33.48) ng/mL, and the concentration with administration at 1 h, 5 h and 10 h was (110 400.00±1 766.35) ng/mL, (1 479.00±687.01) ng/mL and (133.60±12.17) ng/mL, respectively. The results of exosome detection showed that no peak GHB signal was detected in the control group and the 10 h administration group, and the concentrations of GHB at 1 h and 5 h administration groups were (91.47±33.44) ng/mL and (49.43±7.05) ng/mL, respectively. CONCLUSIONS: GHB was detected in blood exosome by UPLC-MS, which indicated that exogenous GHB could be detected in plasma exosomes, while endogenous GHB could not be detected, suggesting that this method may be used as a basis to determine whether there is exogenous drug intake.

MiR-223/PAX6 Axis Regulates Glioblastoma Stem Cell Proliferation and the Chemo Resistance to TMZ via Regulating PI3K/Akt Pathway.

Chemotherapy is a standard strategy for glioma, while chemoresistance remains a major therapeutic challenge in current clinical practice. Our present study was aimed to determine whether inhibition of the miR-223/paired box 6 (PAX6) pathway could increase the sensitivity of glioma to Temozolomide. An elevated level of miR-223 was observed in glioma tissues. Exogenous miR-223 promoted cell survival when exposed to Temozolomide (TMZ), while miR-223 inhibition could reverse this process. The RNA and protein levels of PAX6 were significantly decreased by exogenous miR-223, and the 3'-untranslated region of PAX6 was shown to be a target of miR-223. Besides, it has also been reported that PI3K/Akt signaling pathway is pivotal to regulate glioma growth and proliferation. In the present study, we revealed that miR-223/PAX6 axis regulated the growth, invasion, and chemo resistance of glioblastoma stem cells to TMZ via regulating PI3K/Akt signaling pathway, which present a novel potential therapy for intervention of glioblastoma. Taken together, our findings shed new light on the miR-223/PAX6 pathway in glioma and this pathway might modulate the sensitivity of glioma to TMZ via regulating PI3K/Akt signaling pathway. J. Cell. Biochem. 118: 3452-3461, 2017. © 2017 Wiley Periodicals, Inc.

Gallbladder carcinosarcoma with a poor prognosis: A case report.

BACKGROUND: Carcinosarcoma of the gallbladder is a rare malignant tumor with a very poor prognosis. To date, only approximately 100 patients have been reported in the English literature. The prognosis of this tumor type is poor, the preoperative diagnosis is difficult, and there is a possibility of a misdiagnosis. We present an unsuccessful case of carcinosarcoma of the gallbladder with a preoperative misdiagnosis and rapid early postoperative recurrence. Therefore, we have a deeper understanding of the poor prognosis of gallbladder carcinosarcoma (GBC) patients. CASE SUMMARY: The patient is a 65-year-old male. He was admitted to the hospital because of right upper abdomen distending pain and discomfort for half a month. Abdominal magnetic resonance imaging revealed a polycystic mass in the right lobe of the liver and the fossa of the gallbladder. After admission, the patient was diagnosed with a liver abscess, which was treated by abscess puncture drainage. Obviously, this treatment was unsuccessful. Hepatectomy and cholecystectomy were performed one month after the puncture. Postoperative pathologic examination revealed carcinosarcoma of the gallbladder, and the resected specimen contained two tumor components. One month after surgery, the patient's tumor recurred in situ and started to compress the duodenum, resulting in duodenal obstruction and bleeding. The treatment was not effective. The patient died of gastrointestinal hemorrhage and hypovolemic shock. CONCLUSION: Carcinosarcoma of the gallbladder is a rare malignant tumor that is easily misdiagnosed preoperatively and has a poor prognosis.

Relation between proteinuria and acute kidney injury in patients with severe burns.

INTRODUCTION: Proteinuria in burn patients is common, and may be associated with acute kidney injury (AKI) and adverse outcomes. We evaluated the incidences, outcomes, characteristics and determinants of proteinuria and its influence on AKI and outcomes in burn patients. METHODS: This retrospective study was carried out in a hospital's burn department. The study population consisted of patients with burn injuries admitted during a five-year period. Positive urine dipstick readings were defined as mild (± or 1+) or heavy (≥ 2+) proteinuria, and AKI was diagnosed and staged according to the Risk, Injury, Failure, Loss, End Stage (RIFLE) classification system. Patient characteristics, management and outcomes were evaluated for associations with proteinuria using nonparametric tests, chi-square (χ(2)) tests and binary logistic regression. RESULTS: Of the patients admitted to the burn unit during the study period (n = 2,497), 865 (34.64%) were classified as having proteinuria. In the patients whose total burn surface areas (TBSA) were > 30% (n = 396), 271 patients (68.43%) had proteinuria and 152 of these patients (56.09%) met AKI criteria. No patients without proteinuria developed AKI. Intensive care unit (ICU) mortality rates were 0.8%, 16.67% and 30.77% (P < 0.001) in the groups with no, mild and heavy proteinuria, respectively. Logistic regression analysis identified proteinuria (OR 4.48; 95% CI, 2.824 to 7.108; P < 0.001) and sequential organ failure assessment (OR 1.383; 95% CI, 1.267 to 1.509; P < 0.001) as risk factors for AKI. CONCLUSIONS: We observed a high prevalence of proteinuria in patients with severe burns (> 30% TBSA). Severely burned patients with proteinuria had a high risk of developing AKI and a poor prognosis for survival. This suggests that proteinuria should be used for identifying burn patients at risk of developing AKI.

[Amino acids 395-416 in DNA binding domain of STAT4 is involved in IL-12-induced nuclear import of STAT4].

The purpose of the present study is to explore the mechanism of IL-12-induced nuclear import of signal transducer and activator of transcription 4 (STAT4). Assayed by analyses of homology alignment of STATs, amino acids 395-416 in DNA binding domain was found to be a potential dimer-specific nuclear localization signal (dsNLS) of STAT4. Therefore, several plasmids were constructed. Wild-type STAT4 was inserted into the SalI and BamHI sites of pEGFP-C1 for the construction of plasmid pEGFP-STAT4. The DNA fragment of STAT4 with the deletion of amino acids 395-416 was amplified by RCR and introduced into the SalI and BamHI sites of pEGFP-C1 which was named pEGFP-STAT4-Del. Classic NLS DNA sequence of SV40 T antigen was inserted into the XhoI and HindIII sites of pEGFP-C1. This plasmid was named as pEGFP-NLS and used as a positive control. Plasmid pEGFP-NLS-STAT4-Del was constructed by inserting STAT4-Del into SalI and BamHI sites of pEGFP-NLS. These plasmids were transiently transfected into Caski cells, respectively. The results showed that, after these transfected cells were stimulated by IL-12, wild type STAT4 existed in the cytoplasm at 0 min, and was predominantly localized to the nucleus at 45 min, and distributed in both cytoplasm and nucleus at 60 min, suggesting that STAT4 translocates from cytoplasm into nucleus and finally re-entries into the cytoplasm during the stimulation of IL-12. However, deletion mutant of STAT4 was arrested in cytoplasm during the IL-12 stimulation. Leptomycin B, which specifically blocks protein export from nucleus into cytoplasm, was used to further demonstrate whether STAT4-Del is transferred into nucleus even with stimulation of IL-12. After the transfected cells were pre-treated by leptomycin B, the wild type STAT4 was mainly localized in nucleus after the IL-12 stimulation, suggesting that STAT4 was translocated from cytoplasm into nucleus by the stimulation of IL-12. On the other hand, the deletion mutant of STAT4 distributed in cytoplasm throughout, implying that the mutant STAT4 lacking of amino acids 395-416 cannot move into nucleus. Furthermore, the insertion of classic NLS into EGFP-STAT4-Del restored nuclear import of STAT4-Del. These results suggest the amino acids 395-416 is a dsNLS mediating IL-12-stimulated nuclear import of activated STAT4.

Fatal poisoning by accidental ingestion of the "heartbreak grass" (Gelsemium elegans) verified by toxicological and medico-legal analyses.

We present a case of fatal poisoning from accidental ingestion of Gelsemium elegans (G. elegans), a rarely toxic plant. A 41-year-old man was found dead, at his home, 6 h after drinking homemade herbal liqueur during lunch. Autopsy and routine toxicological analyses identified neither significant pathological findings nor routine poisons. However, a local botanist revealed that the homemade herbal liqueur contained G. elegans, a poisonous plant specific to Asia. To ascertain whether the decedent had ingested G. elegans, we performed liquid chromatography-mass spectrometry (LC-MS) and found two alkaloids (gelsemine and koumine) in his blood, gastric contents, as well as the suspected herbal liqueur. The cause of death was therefore confirmed to be G. elegans poisoning. Case reports of fatal poisoning due to ingestion of G. elegans are quite rare in English. Therefore, the present case broadens the scope on the possibility of death due to ingestion of G. elegans for forensic pathologists and toxicologists.

[Qualitative analysis of the main chemical constituents of shuanghuanglian injection powder and their origin by HPLC-ESI/MS/MS spectrometry].

This paper is to report the analysis of the main chemical constituents of Shuanghuanglian injection powder and determination of their origin. The sample solution was analyzed by a Zorbax C18 column with a gradient mobile phase comprised of methanol and 0.25% acetic acid solution. Both UV and electrospray ionization mass spectrometry detector were used simultaneously, -Q1-scan detection mode was evaluated for the identification of the LC peaks. To analyze the mass spectrum of every LC peaks, 43 molecular mass from the ion chromatogram of Shuanghuanglian injection powder were identified and among them, structure of 20 compounds were elucidated, and the data were sorted to the three component herbs, separately.

[Quantitative determination of 1-deoxynojirimycin in mulberry leaves by high-performance liquid chromatographic-tandem mass/mass spectrometry].

OBJECTIVE: To develop a high-performance liquid chromatographic-tandem mass/mass spectrometric method to determine the concentration of 1-deoxynojirimycin (1-DNJ) in mulberry leaves. METHODS: 1-deoxynojirimycin was separated on an SHIMADZU HRC-NH2 column with the mobile phase consisting of acetonitrile and 0.1% formic acid. The mass spectrometric system equipped with a atmospheric pressure chemical ionization (APCI) interface was operated in Multiple Reaction Monitoring (MRM) mode. RESULTS: The retention time of 1-deoxynojirimycin was 2.87 min, and the calibration curve was linear over a concentration range from 482 microg/L to 2410 microg/L, the average recovery was 95.8%. The detection limit was 53.6 microg/L. CONCLUSION: The method is selective and sensitive for determining 1-deoxynojirimycin in mulberry leaves.

[Study on the chemical constituents of Sarcandra glabra by HPLC-ESI-MS/MS].

OBJECTIVE: To study and compare the main chemical constituents of Sarcandra glabra and qingrexiaoyanning capsules which were extracted by acetic ether. METHODS: The sample solution were analyzed by a Zorbax C18 column with a gradient mobile phase composed of acetonitrile and 0.1% formic acid solution. Both UV and mass spectrometry detector were used simutaneously, full-scan detection mode was evaluated for the identification of all LC peaks. RESULTS: We analyzed the mass spectrum of every LC peak and identified 26 molecular mass from the ion chromatogram of Sarcandra glabra extraction and 16 molecular mass from the extractions of qingrexiaoyanning capsule. 5 compounds were identified. CONCLUSION: High performance liquid chromatography-mass/mass spectrometry has special advantages on analyzing the chemical constituents of traditional Chinese medicine.

Identification of the first case of SFTSV infection in the Hunan Province of China and epidemiological surveillance in the locality.

This study reports the etiological identification, clinical diagnosis, and the results of the local epidemiological surveillance of the first case of severe fever with thrombocytopenia syndrome virus (SFTSV) infection in 2014 in Hunan Province, China. The infected patient was isolated and closely monitored. The virus is a member of the Bunyaviridae sandfly family and is characterized by real-time PCR, electron microscopy, immunofluorescence, and whole-genome sequencing. We also detected IgG and IgM antibodies against SFTSV among the local human population and domestic animals in a serological surveillance. Prevalence of SFTSV-specific antibodies was monitored in the local population for two years after the identification of the first SFTS case. Approximately 5% (4/77) of the people who had direct contact with the patient were seropositive, which is significantly higher than the seropositivity of the general local population [1.57% (44/2800), P < 0.05]. Furthermore, the percentage of the general population who were seropositive was higher in 2015 than in 2014 (χ2 = 7.481, P = 0.006). The epidemiological investigation found that the SFTSV is epidemic in goats, cattle, and chickens in Hunan Province. The risk of infection of domestic animals can be minimized by feeding in pens rather than allowing foraging.

Correlation between expression of cyclooxygenase-2 and the presence of CD4+ infiltrating T-lymphocyte in human primary hepatocellular carcinoma.

BACKGROUND/AIMS: Recent studies have found that Cyclooxygenase-2(Cox-2) is frequently inappropriately expressed in primary hepatocellular carcinoma (HCC), suggesting that abnormal Cox-2 expression plays an important role in hepatocarcinogenesis. But it remains controversial in these reports. Moreover, there are only a few studies on the correlation between Cox-2 and infiltrating immunocytes in the tumor-microenvironment. CD4+ tumor-infiltrating lymphocytes (TIL) and infiltrating immunocytes around the tumor are closely correlated to the development of the tumor, but so far no reports are available showing the relationship among Cox-2, CD4+ TIL of tumor and CD4+ infiltrating T-lymphocytes of adjoining non-tumorous (ANT) tissues in tumor-microenvironment. This study is designed to appropriately select and collect patients' specimens to better reflect Cox-2 expression in human HCC, and also to stress the correlation among Cox-2, CD4+ TIL and CD4+ infiltrating T-lymphocytes in the tumor-microenvironment. METHODOLOGY: Tumor tissue, and its matched ANT tissue removed less than 1 cm from the solid tumor border, were obtained from 25 HCC patients all of whom came from Hunan province, China, and were infected with hepatitis B virus (HBV). Normal liver tissues of 10 hemangiomas were collected as controls. Both Cox-2 expression and the number of CD4+ TIL and CD4+ infiltrating T-lymphocyte were detected by immunohistochemistry, and data were analyzed closely with patients' clinical figures so as to investigate the correlation between the 3 elements. RESULTS: In 25 HCC patients, remarkably higher Cox-2 expression in both tumor and ANT tissues was observed compared with normal liver tissues (p < 0.001). The percentage of Cox-2 positive cells was, remarkably, higher in ANT tissues than in tumors (p < 0.001). Similarly the distribution of CD4+ T cells was significantly higher in ANT tissue than in tumor tissue (p < 0.0001), and also significantly higher in tumor tissue than in normal tissue (p < 0.0001). Importantly, in the group of patients with Cox-2-expressing tumors, the number of CD4+ infiltrating T-lymphocyte in ANT tissues was 79.4(+)9.92/hpf, which is obviously lower (p = 0.019) than that of the group with non-Cox-2-expressing tumors with the number of CD4+ infiltrating T-lymphocyte in ANT tissues at 118.13(+)12.47/hpf. Cox-2 expression of tumors showed a significant negative correlation with number of CD4+ infiltrating T cells of ANT tissues (r = 0.499, p = 0.024). The number of CD4+ TILs are lower in Cox-2-expressing tumors than in non-Cox-2-expressing tumors, but there was not statistical significance (p = 0.057). CONCLUSIONS: Taken together we suggest in the tumor-microenvironment of HCC the expression of Cox-2 may inhibit the number CD4+ infiltrating T-lymphocyte in ANT tissues. As a result, Cox-2 overexpression may contribute to both suppression of local immune responses and enhancement of metastatic potential in human HCC.

[MICA*008/A5.1 allele and HCMV infection in kidney transplanted donees of Hunan Han nationality].

OBJECTIVE: To investigate the relationship between MICA*008/A5.1 allele and human cytomegalovirus (HCMV) infection in kidney transplanted donees of Hunan Han nationality. METHODS: The MICA*008/A5.1 allele based on 91 kidney transplanted donees and 81 unrelated normal individuals of Han nationality in Hunan Province were analyzed by PCR/SSP assay. At the same time, anti-HCMV antibody IgM was detected in the serum by ELISA method. RESULTS: The positive rate of MICA*008/A5.1 allele was significantly higher in the control group (56.79%) than that in the kidney transplanted donee group (34.07%) (P <0.05). The infection rate of HCMV in those individuals whose genotype was MICA*008/A5.1 (-) was significantly higher than that in the MICA*008/A5.1(+). CONCLUSION: The individual whose genotype is MICA*008/A5.1 (+) is not liable to HCMV infection, but the individual whose genotype is MICA*008/A5.1 (-) is liable to HCMV infection.

[Polymorphism of short tandem repeat of exon 5 of MHC class-I chain related gene A and association with nasopharyngeal carcinoma in a southern Chinese population].

OBJECTIVE: To explore the association between the short tandem repeat polymorphism of exon 5 of MICA gene (MICA-STR) and nasopharyngeal carcinoma (NPC) in a southern Chinese population. METHODS: One hundred and twenty-seven consecutive NPC patients and 112 randomly selected normal controls residing in southern China mainland were analyzed for MICA-STR allelic variation and MICA gene deletion by fluorescent polymerase chain reaction-gene scanning and polymerase chain reaction-sequence specific priming. RESULTS: MICA*A9 was observed at significantly higher frequency in the NPC patient group than in the control group (relative risk = 2.524, P = 0.001,Pc = 0.006); whereas MICA*A5.1 was present at significantly lower frequency in the NPC patient group than in the control group (RR = 0.418, P = 0.0004, Pc = 0.0026). Further analysis revealed that MICA*A9 was over-represented in male NPC patients, compared with male controls (RR = 3.23, P = 0.00095, Pc = 0.006); whereas MICA*A5.1 was present at significantly lower frequency in male NPC patients, compared with male controls (RR = 0.372, P = 0.0007, Pc = 0.004). None of the MICA-STR variants showed statistically significant frequency difference between female NPC patients and female controls (Pc > 0.05). CONCLUSION: MICA-STR polymorphism is associated with NPC, and MICA*A9 is a genetic susceptibility marker of male individuals for NPC in a southern Chinese population.

[High-performance liquid chromatography combined with mass spectrum analysis for chromatographic fingerprinting of compound dandelion enema].

OBJECTIVE: To identify the active components of compound dandelion enema, a preparation from 7 traditional Chinese herbal drugs for treatment of gynecological diseases. METHODS: Three-dimensional high-performance liquid chromatography (3D-HPLC) was employed to separate the ethyl acetate extract of compound dandelion Enema, and HPLC combined with mass spectrum (MS) analysis used for chromatographic fingerprinting. RESULT: By comparing the ionic fragments of MS and retention time of each peak, the main active components in compound dandelion enema were determined, including caffeic acid, ferulic acid and protocatechualdehyde. CONCLUSION: HPLC coupled with mass spectroscopy can be used for qualitative analysis of compound dandelion enema.

Berberine Increases Doxorubicin Sensitivity by Suppressing STAT3 in Lung Cancer.

Berberine (BBR), an alkaloid component isolated from Chinese medicinal herb Huang Lian, has aroused broad interests for its antitumor effect in recent years. The signal transducer and activator of transcription 3 (STAT3), plays critical roles in malignant transformation and progression and was found to be constitutively activated in a variety of human cancers. In this study, we show that BBR inhibited cell proliferation, induced apoptosis, and suppressed tumor spheroid formation of lung cancer cell lines. These effects were correlated with BBR-mediated suppression of both phosphorylated and total levels of STAT3 protein. Furthermore, BBR promoted STAT3 degradation by enhancing ubiquitination. Importantly, we demonstrated that BBR was able to inhibit doxorubicin (DOX)-mediated STAT3 activation and sensitize lung cancer cells to the cytotoxic effect of DOX treatment. Given that BBR is widely used in clinic with low toxicity, our results are potentially important for the development of a novel combinatorial therapy with BBR and DOX in the treatment of lung cancer.

[Studies on design, synthesis and biodegradation of carrier for colon-site specific, drug delivery system].

AIM: To design and synthesize a novel vector for colon-site specific drug delivery system and investigate the relationship between the biodegradation properties and composition of materials in the simulated colon fluid. METHODS: The azocopolymer P (HEMA-MMA-MAA) was synthesized using 2-hydroxyethylmethacrylate (HEMA), methyl methacrylate (MMA) and methacrylic acid (MAA) as comonmer, azobisisobutyronitrilel (AIBN) as initiator, cross-linked with divinylazobezene (DVAB). The chemical structure of the synthesized series of azocopolymer is examined by UV, FTIR spectroscopy and nuclear magnetic resonance data. Their swelling behavior is evaluated by the swelling equilibrium parameter Q, the biodegradation tests of the materials were carried out at physiologically relevant buffer designed to mimic the colon environment. The biodegradation properties were assessed using the differential scanning calorimeters (DSC) and gel permeation chromatography (GPC) and the morphology on the surface of materials before and after degradation was observed by scanning electron microscopy (SEM). RESULTS: The swelling equilibrium parameter Q increased with increasing the contents of HEMA and MAA in the materials. The degradation behavior was relevant to the ratio of three components in the copolymers. CONCLUSION: This materials may become a good carrier for the colon-site specific drug delivery system if the contents of commoners HEMA, MMA and MAA are adjusted reasonably.

[High-performance capillary electrophoresis for determination of urinary citrate and oxalate].

OBJECTIVE: To develop a method for determining urinary citrate and oxalate using high-performance capillary electrophoresis. METHODS: Capillary electrophoresis with indirect ultraviolet (UV) detection was performed for determining urinary citrate and oxalate. Sodium hromate was used as the UV-absorbing background electrolyte. RESULTS: Citrate and oxalate had a migration time of 3.76 min and 3.14 min respectively. The calibration curves were linear within a limit of about 2 microg/ml for citrate and about 1 microg/ml for oxalate. The recovery rates of citrate and oxalate were 97.53% and 99.11% respectively. CONCLUSION: This method is simple, accurate and at low cost for determining citrate and oxalate in the urine.

[Determination of the total alkaloid content of Rhizoma coptidis in Gegenqinlian prescription].

OBJECTIVE: To determine the total content of alkaloids from Rhizoma coptidis in the traditional Chinese medicine prescription Gegenqinlian. METHODS: The extract of Gegenqinlian decoction and powder of the minipills were respectively purified with alumina column chromatography, and the total alkaloid content of Rhizoma coptidis in these preparations was determined using ultraviolet spectrophotometry at 350 nm. RESULTS: The total alkaloid content was 32.4 mg/g in the aqueous extract of Gegenqinlian decoction and 19.6 mg/g in Gegenqinlian minipills, with a recovery rate of 98.1% (RSD=2.9%). CONCLUSION: The method we employed yields accurate and stable results with much sensitivity for quality control of Gegenqinlian minipills, and also provides reference for study of Gegenqinlian prescription and other prescriptions that contain Rhizoma coptidis.

High-performance capillary electrophoresis for determining the contents of berberine, jatrorrhizine and palmatine in Gegenqinlian decoction.

OBJECTIVE: To develop a method for alkaloid isolation from the extract of Gegenqinlian decoction (a traditional Chinese herbal preparation) and content determination. METHOD: After the preparation of the Gegenqinlian decoction extract and the micropellets, the quaternary alkaloids of Coptis chinensis were isolated by high-performance capillary electrophoresis (HPCE) in a buffer solution containing 60% sodium phosphate (60 mmol/L, pH 8.0) and 40% methanol. 2-(4-hydroxyphenyl) ethylammonium chloride was used as the internal standard and the ultraviolet detection was performed at 254 nm for determining the contents of berberine, jatrorrhizine and palmatine in the extract of Gegenqinlian decoction. RESULTS: Berberine, jatrorrhizine and palmatine were successfully isolated within 6 min with recovery rates of 99.05%, 98.70% and 97.76% respectively and relative standard deviation of 2.12%, 1.85% and 1.95% respectively. CONCLUSION: HPCE is applicable for the determination and isolation of quaternary alkaloids in Coptis chinensis with high accuracy and recovery rate.