Absorption is the movement of a drug from its site of administration to the
systemic circulation by crossing biological membrane. The absorption study of certain drug varies as per the route of absorption used. Absorption study can be performed mainly by two methods: 1. In vivo study 2. In vitro study 1. In vivo study In vivo study can be performed in either of the following: a) Animals (Non clinical study) b) Human beings (Clinical study) In vivo study in animals (Non clinical study) Different animals can be selected for non-clinical study. For the percutaneous invivo absorption study of Retinol, a kind of rat, Female fuzzy rats were used. The rats of age 3-14 months were selected which must be free of any sort of diseases. The rats are provided with proper food, water and are kept in a properly maintained cage (211 C with 5010% relative humidity and a twelve-hour light/dark full spectrum lighting cycle with no twilight). For percutaneous in-vivo absorption study of retinol, an area of application of 3.03.0 cm was delineated on the mid-scapular region of the rat. Rats were dosed and the patching were enclosed and protected with a Stomahesive patch glued directly to the animal's skin. The animals were placed in metabolism cages for 24 or 72 h to collect urine and faeces. The treated skin site was washed 24 h after application with soap and water to remove unabsorbed material. The washes were collected to determine the amount of unabsorbed material. The skin dosing site and remaining carcass were each dissolved in concentrated (5 M) potassium hydroxide. Aliquots of the urine, feces, and dissolved carcass were analyzed for radioactivity by liquid scintillation counting. The distribution of radioactivity was determined in the area of the dosing site. The skin with the patch still intact was cleaned of fat from the underlying tissue. The patch was then removed and the remaining glue on the skin was removed with hexanesoaked cotton swabs. The area containing the dosing site was punched into one or two skindiscs. Each skin disc was tape stripped 10-times to determine the amount of retinol remaining in the stratum corneum vs. the epidermis/dermis.
The absorption values were then analyzed.
In-vivo study in human (Clinical study) For clinical study, different healthy and target diseased human volunteers are selected. For Paracetamol absorption study, 12 acute stroke patients and 13 healthy contols were selected of similar age. They were given the dose of 1g oral paracetamol tablets. 10 ml venous blood samples were collected at 15 minutes interval for 4 hours for paracetamol measurement. The samples were centrifuged and the separated plasma was stored at -20C until the completion of the analysis. The analysis was performed by reverse phase HPLC. The pharmacokinetic measurements Tmax and Cmax were measured from plasma concentration time profile, AUC was measured by trapezoidal rule and the results in the volunteers were compared. 2. In vitro study In vitro absorption studies are performed without the use of live animals or human beings. For percutaneous absorption study of Retinol, rat skin and human skin sections were used. A split-thickness layer of rat or human skin (200320 m) was prepared with a dermatome and discs of dermatomed skin were obtained using a brass punch (13/16 in. diameter). Each skin disc was then mounted with the epidermal side up in the diffusion cell. The retinol dose (2 mg/cm2 application amount) was applied to each diffusion cell for 24 h, and then washed off to remove any unabsorbed material. Amounts remaining in the skin and absorbed into the receptor fluid were determined.At 24 h after dosing, the retinol remaining on top of the skin as unabsorbed material was removed by washing three times with 0.1 ml of a 10% (v/v) liquid detergent solution that was pipetted onto the skin surface. The skin surface was gently rubbed with cotton-tipped swabs to remove the detergent solution. The skin was rinsed two times with 0.2 ml of distilled water. All cotton swab tips were collected in a scintillation vial as a measure of the amount of material remaining on the skin surface. At the end of the study (24 or 72 h), the skin was removed from the diffusion cell and the amount of retinol remaining in the skin was determined. Conclusion Hence, in vivo absorption study in any animals or human volunteers and in vitro absorption study using suitable method helps to estimate systemic absorption of any drugs.