UNICORN GH+ from Beacon Group

Chromatography is a biophysical technique for separation, identification, and purification of target molecules from a complex mixture for qualitative and quantitative analysis. Components like stationary phase, mobile phase, and separated molecules form the basis of chromatography techniques. Stationary phase is quoted on solid surface while mobile phase could be either gas or liquid phase. If the mobile phase is liquid, it is generally termed as liquid chromatography.

HPLC represents the closest approach to an ideal chromatographic system. Dense column packaging, pressure-metering system, and use of on-line detector provide more superior separation. For several years, HPLC has been largely used in pure chemistry, and by the pharmaceutical industry for convenient monitoring of chemical reactions as well as to check the assessment of purity of compound mixtures used in reactions.

Based on the superiority of chromatographic characterization, in the recent past, HPLC has been used in clinical chemistry laboratories due to its abilities to separate and compare the molecule to a higher magnitude.

HPLC is suitable for a variety of clinical applications, including catecholamines measurements, vitamin monitoring, and detecting presence of illicit drugs from biological samples. In addition to these, HPLC is more frequently used for accurate diagnosis of conditions like hemoglobino­pathies and thalassemia, and to measure glycosylated hemoglobin.

Clinical laboratories routinely handle hundreds of samples, such as blood, urine, and other body fluids. Efficient extraction processes are required to be coupled with rapid analysis to handle such heavy loads but without sacrificing quality of analysis because patient’s treatment by medical doctors is based on such reports.

The advantage of the HPLC system is the excellent resolution, reproducibility, and quantification of several normal and abnormal analytes of clinical interest, resulting in accurate diagnosis in a timely manner.

Advances in column-packing materials technology, such as sub-micron superficially porous silica particles introduced the ultra-high pressure HPLC which improved separation efficiencies without encountering high backpressures. Introduction of micro-bore columns offered faster analysis times. A combination of other advances, such as efficient autosamplers and reduced detector cell volumes, and improvements in data collection and analysis contributed significantly to high sample throughputs in clinical laboratories. This is a key requirement in such laboratories, which need to generate reports on hundreds of samples for multiple parameters on daily basis.

Approximately 387 million people are living with diabetes worldwide, with an estimated prevalence of 8.3 percent in 2014, which is expected to reach 10 percent by 2030. According to a survey of 2015, 30.3 million people or 9.4 percent of total population of America were diabetic. On the other hand, 77 million people are found diabetic in recent survey of IDF in India.

Since 1958, when HbA1c was first separated from other forms of haemoglobin, it has been considered the gold standard for the monitoring of glucose control in patients with diabetes. HPLC provides signi­ficant and unique benefits for monitoring of HbA1c. In today’s modern laboratories, HPLC for HbA1c can be fully integrated on automation platforms. This enables labs to provide more reliable gold standard results with ease of use. Moreover, HPLC has distinct advantage of being able to identify variants in their hetero­zygous state as well as FHb, CHb, and LA1c.

Ion exchange HPLC, working on the basis of separation by molecular charges, can effectively separate glycated Hb from non-glycated ones in a shorter time and with inter run or intra run CV of >2 percent. By this method, even more common Hb variants can also be detected.

UNICORN GH+, added into the product portfolio of Beacon Group recently, is a perfect solution for HbA1c monitoring by HPLC method. Using three-gradient elution method, coupled with specially designed optimized high-pressure column and double plunger reciprocating pump system, UNICORN GH+ provides results in >3 min with measuring range up to 18 percent.

HPLC is surely offering enormous advantages to clinical labs for effective separation of target molecule from complex biological samples for efficient diagnosis and prognosis.