OBJECTIVETo develop a new method to rapidly determine and identify Guizhi Fuling capsule by portable acousto-optic tunable filter-near infrared spectroscopy.

METHODThe qualitative model was set up using principal component analysis. The correlation models between the NIR spectra and the reference values of five major constituents were obtained with partial least squares method.

RESULTThe identifying model accurately identified Guizhi Fuling capsule, and quantitative analytical models could precisely predicted the content of ellagic acid, baicalin, benzoylpaenoniflorin, cinnamaldehyde, and paeonol. The correlation coefficients of the calibration models were 0.924 2, 0.938 4, 0.924 2, 0.933 6, 0.934 7, the validation set coefficients of the calibration were 0.924 2, 0.938 4, 0.924 2, 0.933 6, 0.934 7, and the RMSEP were 1.138%, 3.014%, 0.751%, 0.625%, 3.455%, 1.363%, respectively. The results of external validation showed no significant difference between the predictive and the determining values by t-test.

CONCLUSIONThe method is accurate, rapid and non-destructive, and can be used for determining and identifying Guizhi Fuling capsule.

Acetophenones ; analysis ; Acrolein ; analogs & derivatives ; analysis ; Calibration ; Capsules ; chemistry ; Drug Evaluation ; methods ; Drugs, Chinese Herbal ; analysis ; Ellagic Acid ; analysis ; Flavonoids ; analysis ; Least-Squares Analysis ; Models, Chemical ; Principal Component Analysis ; methods ; Spectroscopy, Near-Infrared ; methods

OBJECTIVE： To screen active fractions of Xiaozhong zhitong lotion that are able to reduce swelling， promote ulcer healing and analgesia， and to provide reference for it’s secondary development of ointment preparation. METHODS： Water elution fraction and 20％， 40％， 60％， 95％ ethanol elution parts were separated by D101 macroporous resin from Xiaozhong zhitong lotion. 120 SD rats were randomly divided into normal group （n＝8） and modeling group （n＝112）. Rats in the normal group were not treated. Hemorrhoids model was established in the model group by injecting 75％ glacial acetic acid into the perianal skin to induce perianal ulcer. 96 model rats were randomly divided into model group [blank ointment matrix 0.51 g/（kg·d）]， positive group [Mayinglong ointment， 0.51 g/（kg·d）]， high-dose and low-dose groups of Xiaozhong zhitong lotion and it’s water elution fraction and 20％， 40％， 60％ ethanol elution parts [8.34， 2.78 g/（kg·d） by crude drug， all make into containing drug ointment]， with 8 rats in each group. The periphery of the anus was smeared with relevant medicine， twice a day， for consecutive 7 d. The local symptoms around the anus of rats 3 and 7 days after administration and the pathological morphology of the local mucosa around the anus of rats 7 days after administration were observed and scored respectively. The effects of each elution part for reducing swelling and promoting ulcer healing were investigated. 120 ICR mice were randomly divided into model group [blank ointment matrix 1.03 g/（kg·d）]， positive group [Mayinglong ointment 1.03 g/（kg·d）]， high-dose and low-dose groups of Xiaozhong zhitong lotion and it’s each elution part [16.65， 5.55 g/（kg·d） by crude drug， all make into containing drug ointment]， with 10 mice in each group. Transdermal administration， twice a day， for consecutive 7 d. After 30 min of last administration， the latency time and 15 min writhing times of mice were detected by designing acetic acid writhing test； pain threshold of mice was determined by hot-plate pain test so as to investigate systemic and local analgesic effects of each elution part. RESULTS： In the detumescence and ulcer healing test， compared with normal group， the score of local symptoms around anus at 3rd and 7th day of administration as well as pathological score of hemorrhoids local mucosa were increased significantly in model group （P＜0.01）. Compared with model group， above scores of positive group， 40％ ethanol elution high-dose and low-dose groups were decreased significantly （P＜0.05 or P＜0.01）. The scores of local symptoms around anus in 20％ ethanol elution part high-dose group at 3rd and 7th day after medication as well as 60％ ethanol elution part group at 7th day after medication were decreased significantly （P＜0.05）. In analgesia test， compared with model group， writhing latency time was shortened significantly and 15 min writhing times was decreased significantly in positive group， 40％ ethanol elution part high-dose and low-dose groups as weel as 60％ ethanol elution part high-dose group （P＜0.05 or P＜0.01）； writhing latency time of 20％ ethanol elution part high-dose group was shortened significantly （P＜0.05）. Pain threshold of mice was increased significantly in positive group， 40％ ethanol elution part high-dose and low-dose groups after medication （P＜0.05 or P＜0.01）. CONCLUSIONS： The 40％ ethanol elution part from Xiaozhong zhitong lotion is the effective part that can reduce swelling， promote ulcer healing and analgesia.

OBJECTIVE： To analyze chemical components in active fraction of Xiaozhong zhitong lotion， to clarify the material basis of its efficacy， and to provide reference for the second development of ointment preparation. METHODS： UPLC-Q-TOF-MS was adopted to analyze the chemical components of active fraction （40％ ethanol elution site separated by D101 macroporous resin） of Xiaozhong zhitong lotion. The determination was performed on Hypersil GOLD aQ C18 column with mobile phase consisted of 0.1％ formic acid acetonitrile （A）-0.1％ formic acid water （B） （gradient elution） at the flow rate of 0.4       mL/min. The sample size was 4 μL， and the column temperature was 30 ℃. The condition of mass spectrometry was ESI detection in positive and negative scanning ion mode （ESI+/ESI－）. The scanning range was 100-2 000 Da. The collision energy was 45/－45 eV， and the energy of the extended collider was 10/15 eV. The accurate molecular weight， retention time and multi-stage fragment ion information of the compounds were collected after obtaining the chromatogram， and the chemical components were identified by comparing with the mass spectrum information of reference materials and references. RESULTS： A total of 48 compounds were identified， and 9 and 39 compounds were identified under ESI+/ESI－ ion mode， mainly including 10 phenolic acids， 8  phenylpropanoids， 9 anthraquinones， 3 flavones， 7 alkaloids， 5 tannins and 6 other categories. CONCLUSIONS： UPLC-Q-TOF- MS method is rapid， efficient and accurate for identify chemical components from active fraction of Xiaozhong zhitong lotion. Main chemical components of the active fraction are phenolic acids， phenylpropanoids， anthraquinones， alkaloids and tannins.