OBJECTIVETo develop an RP-HPLC method for assay of pilocarpine in rabbit ocular aqueous humor.

METHODSThe RP-HPLC method was performed on a column of ODS-C(18) with the mobile phase consisting of 0.5% of triethylamine (TEA) of phosphate solutions (10 mmol/L, pH 2.5) and acetonitrile (98/2,v/v). The detection wavelength was 215 nm and flow rate was 1.0 ml/min. Ninety albino rabbits were divided into 3 groups (30 in each):group 1 received 50 microl of eye drops containing 1% generic pilocarpine, group 21% mixture pilocarpine solution consisting of aqueous sample and liposome and group 31% liposome pilocarpine, respectively. The aqueous humor was withdrawn at 5, 10, 30, 40, 60, 90, 120, 180, 240 and 360 min. Pilocarpine was extracted from aqueous humor with dichloromethane.

RESULTThe linear calibration curve was obtained in the concentration range of 0.1 - 20 microg/ml. The average recovery was (68.1+/-2.7)% (n=9). Inter-day and intra-day RSD were 4.33% and 2.87%, respectively. In three formations 1% liposome pilocarpine was the best for the areas under curve and measurable amounts.

CONCLUSIONThe RP-HPLC method is simple and reliable for pilocarpine measurement in ocular aqueous. Liposome formulation can significantly increase the bioavailability of pilocarpine in ocular aqueous.

Animals ; Aqueous Humor ; chemistry ; Chemistry, Pharmaceutical ; Chromatography, High Pressure Liquid ; Pilocarpine ; administration & dosage ; analysis ; Rabbits

In this paper we reviewed the formulae and analyzed the factors which modified the relationship and estimate the time of death as precise as possible by potassium concentration in vitreous humor. The extra factors including numbers of the cases, cause of death, different eye of both, the urea or/and creatinine in vitreous humor, environmental temperature, sampling techniques, experimental and analytical method were also mentioned.
Aqueous Humor/chemistry* ; Creatinine/analysis* ; Forensic Medicine ; Humans ; Linear Models ; Postmortem Changes ; Potassium/analysis* ; Time Factors ; Urea/analysis*

A HPLC method was developed for the determination of mangiferin in rat plasma and aqueous humor. 4-Nitrophenol was used as internal standard. Analysis was performed on a Cosmosil ODS C18 analytical column (4.6 mm x 250 mm, 5 microm) with mobile phase consisting of methanol-water (40: 60) with 2% glacial acetic acid at a flow rate of 1.0 mL x min(-1). The calibration curve of mangiferin in rat plasma and aqueous humor showed excellent linear behaviors over the investigated concentration of 0. 50-250.00 mg x L(-1) in plasma and 0.10-10.00 mg x L(-1) in aqueous humor, respectively, and the correlation coefficients were all above 0.995 4. The intra-day and inter-day precisions for all samples were measured to be below 12%. The limit of quantitation was 0.10 mg x L(-1) and low enough for the determination of mangiferin in all samples. The validated method has been successfully applied to preliminary pharmacokinetics study of mangiferin in rat plasma and aqueous humor.
Animals ; Aqueous Humor ; chemistry ; Chromatography, High Pressure Liquid ; Female ; Male ; Nitrophenols ; analysis ; Rats ; Rats, Wistar ; Reproducibility of Results ; Sensitivity and Specificity ; Xanthones ; blood ; pharmacokinetics

OBJECTIVE: To explored the relationship between the concentration of potassium ion as well as sodium ion in the aqueous humor and post-mortem interval (PMI). METHODS: The concentrations of potassium ion and sodium ion in the aqueous humor of swine within 48 h after death at 4 degrees C and 28 degrees C were detected using Z-500 atomic absorption spectrophotometer. RESULTS: The concentrations of potassium ion and sodium ion in aqueous humor of isolated swine eyeballs within 48 h after death when the environmental temperature was 4 degrees C were significantly related to PMI. The relationship between PMI and the concentration of potassium ion was PMI = -0.178[K+]2 + 49.978 (R2 = 0.995). The relationship between PMI and the rate of sodium ion and potassium ion was PMI = 120.987/[Na+/K+]-28.834 (R2 = 0.905). CONCLUSION The concentration of potassium in aqueous humor of isolated swine eyeballs may be one of the reference indicators to estimate PMI of the corpses at lower temperatures.
Animals ; Aqueous Humor/chemistry* ; Female ; Forensic Pathology/methods* ; Male ; Postmortem Changes ; Potassium/analysis* ; Regression Analysis ; Sodium/analysis* ; Spectrophotometry, Atomic/methods* ; Swine ; Temperature ; Time Factors

AIMTo prepare diclofenac sodium liposomes and observe its ocular pharmacokinetics in rabbits.

METHODSThe diclofenac sodium cationic liposomes were prepared by reverse-phase evaporation methods and the formula of liposome was optimized with uniform design. HPLC method was established and validated for the determination of diclofenac sodium in precornea, cornea and aqueous humor of rabbit eye. Liposome and eyedrop solution 50 microL with total 50 microg diclofenac sodium were instilled to eyes of rabbits, separately. Samples of tear, cornea and aqueous humor were collected at different time intervals after rabbits were sacrificed. The ocular pharmacokinetics was investigated by the concentration-time data of tear, cornea and aqueous humor.

RESULTSThe mean particle size of the diclofenac sodium liposomes was 226.5 nm with zeta potential of + 18. 1 mV. The entrapment efficiency reached 63%. Compared with solution, liposome was characterized by slower clearance in precornea. The concentration of diclotenac in cornea and aqueous humor instilled with liposome were higher than that with eye-drop solution. Cmax of diclofenac sodium in aqueous humor instilled with liposome and eye-drop solution were (0.69 +/- 0.25) and (0.48 +/- 0.19) microg x mL(-1) and (36.68 +/- 11.7) and (21.82 +/- 8.6) microg x g(-1) in cornea, respectively. But no significant difference were found to Tmax in aqueous humor and cornea between liposome and eyedrop, T(1/2) of diclofenac in aqueous humor and cornea with liposoine were longer than that with eye-drop solution. The ocular bioavailability of liposome in aqueous humor was 211% compared with that of eyedrop.

CONCLUSIONDiclofenac sodium cationic liposomes can increase the corneal contact time, enhance the corneal permeability of diclofenac sodium and improve its ocular bioavailability.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; chemistry ; pharmacokinetics ; Aqueous Humor ; metabolism ; Area Under Curve ; Biological Availability ; Chromatography, High Pressure Liquid ; Cornea ; metabolism ; Diclofenac ; administration & dosage ; chemistry ; pharmacokinetics ; Drug Compounding ; Liposomes ; Microscopy, Electron, Transmission ; Particle Size ; Rabbits ; Random Allocation

PURPOSE: To compare the levels of calcium and phosphorus in the aqueous humor and serum of diabetics and non-diabetics. METHODS: We divided patients into two groups: seventy-six non-diabetic cataract patients and fifty-two diabetic cataract patients. The diabetic group was divided again into three subgroups: twenty-six patients with no diabetic retinopathy, thirteen patients with non-proliferative diabetic retinopathy, and thirteen patients with proliferative diabetic retinopathy. The authors compared the levels of calcium and phosphorus in the serum and aqueous humor of cataract patients. Statistic analysis was performed to form two comparisons: 1) a comparison between non-diabetics and diabetics and 2) a comparison among non-diabetics and the three subgroups of diabetics. RESULTS: In serum, calcium levels did not statistically differ between non-diabetics and diabetics. The phosphorus level was also not significantly different. In the aqueous humor, however, while calcium levels did not differ significantly, the phosphorus levels in diabetics were considerably higher than those in non-diabetics. When non-diabetics were compared to the three diabetic subgroups, calcium levels did not differ in serum or aqueous humor, but the phosphorus levels in diabetics with proliferative diabetic retinopathy were significantly higher than those in non-diabetics, diabetics without diabetic retinopathy, and diabetics with non-proliferative diabetic retinopathy. CONCLUSIONS: The level of phosphorus in the aqueous humor and serum of diabetics was significantly increased, especially in diabetics with proliferative diabetic retinopathy. This result may be related to hydrophilic acrylic IOL opacification. Future studies regarding the pathogenic role of a high concentration of aqueous humor and serum phosphorus are required.
Aged ; Aged, 80 and over ; Aqueous Humor/*chemistry ; Biological Markers/analysis ; Calcium/*analysis ; Cataract/complications/*metabolism ; Diabetic Retinopathy/complications/*metabolism ; Female ; Humans ; Male ; Middle Aged ; Phosphorus/*analysis ; Regression Analysis ; Retrospective Studies

PURPOSE: To compare the levels of calcium and phosphorus in the aqueous humor and serum of diabetics and non-diabetics. METHODS: We divided patients into two groups: seventy-six non-diabetic cataract patients and fifty-two diabetic cataract patients. The diabetic group was divided again into three subgroups: twenty-six patients with no diabetic retinopathy, thirteen patients with non-proliferative diabetic retinopathy, and thirteen patients with proliferative diabetic retinopathy. The authors compared the levels of calcium and phosphorus in the serum and aqueous humor of cataract patients. Statistic analysis was performed to form two comparisons: 1) a comparison between non-diabetics and diabetics and 2) a comparison among non-diabetics and the three subgroups of diabetics. RESULTS: In serum, calcium levels did not statistically differ between non-diabetics and diabetics. The phosphorus level was also not significantly different. In the aqueous humor, however, while calcium levels did not differ significantly, the phosphorus levels in diabetics were considerably higher than those in non-diabetics. When non-diabetics were compared to the three diabetic subgroups, calcium levels did not differ in serum or aqueous humor, but the phosphorus levels in diabetics with proliferative diabetic retinopathy were significantly higher than those in non-diabetics, diabetics without diabetic retinopathy, and diabetics with non-proliferative diabetic retinopathy. CONCLUSIONS: The level of phosphorus in the aqueous humor and serum of diabetics was significantly increased, especially in diabetics with proliferative diabetic retinopathy. This result may be related to hydrophilic acrylic IOL opacification. Future studies regarding the pathogenic role of a high concentration of aqueous humor and serum phosphorus are required.
Aged ; Aged, 80 and over ; Aqueous Humor/*chemistry ; Biological Markers/analysis ; Calcium/*analysis ; Cataract/complications/*metabolism ; Diabetic Retinopathy/complications/*metabolism ; Female ; Humans ; Male ; Middle Aged ; Phosphorus/*analysis ; Regression Analysis ; Retrospective Studies

OBJECTIVETo detect the levels of vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) in aqueous humor of patients with active choroidal neovascularization (CNV).

METHODSAqueous humor samples were obtained from 32 patients with active CNV. The concentrations of VEGF and PEDF in aqueous humor were measured by enzyme linked immunosorbent assay (ELISA) for quantitative analysis. VEGF and PEDF in 10 samples of aqueous humor from patients with cataract were also detected by the same methods as control.

RESULTThe mean VEGF and PEDF concentrations in aqueous humor of active CNV patients were higher than those in the control group (P=0.000).

CONCLUSIONThe patients with active CNV exhibit significantly higher VEGF and PEDF levels than those in control, indicating that VEGF along with PEDF may modulate the formation of CNV.

Adult ; Aged ; Aged, 80 and over ; Aqueous Humor ; chemistry ; Choroidal Neovascularization ; metabolism ; Eye Proteins ; analysis ; Female ; Humans ; Male ; Middle Aged ; Nerve Growth Factors ; analysis ; Serpins ; analysis ; Vascular Endothelial Growth Factor A ; analysis