High Performance Liquid Chromatography


Introduction

High performance liquid chromatography (HPLC) is an analytical technique used to separate, identify, quantify, and purify individual components of a mixture. This technique is very common in biological and chemical research. Purdue University has numerous HPLC units located in several laboratories around campus. Because organic solvents (e.g., methanol and acetonitrile) are used in the process, HPLC waste is regulated by the Environmental Protection Agency as hazardous waste.


HPLC Waste Collection

Containers collecting HPLC waste must remain closed while the HPLC unit is in operation. It is neither acceptable to place a waste line running from the HPLC unit into an open waste container nor is it acceptable to use foil or Parafilm® as a means of closure (Table 1).

One of the following practices must be employed in order to comply with hazardous waste regulations for HPLC waste collection systems:

  1. Purchase an engineered container and/or cap designed for HPLC waste collection.
    Table 2 shows several examples of acceptable solutions for proper HPLC waste collection that can be purchased.
  2. Modify an existing cap.
    An existing cap can be modified by the research lab for HPLC waste collection. To modify an existing cap, a hole can be drilled into a cap. The diameter of the hole should be similar to the diameter of the waste line; there should be a tight fit between the container opening and waste line. In addition, a hole should be drilled to accommodate any exhaust filter or air valve tube that may be required. It is recommended that either a 4-liter container or 5-gallon carboy be used for waste collection. The modified cap should be replaced with a regular, unmodified cap once the container is full and ready for EHS pickup. See Table 3 for examples of acceptable modified caps.
Table 1: Improper HPLC Waste Collection Practices
Open Waste Open Waste Foil Parafilm®
Four liter brown glass HPLC waste container with hoses going inside. Twenty liter carboy HPLC waste container with hoses going inside. HPLC waste collection container opening with hoses going into it covered by foil. HPLC waste collection container opening with hoses going into it covered by Parafilm.

 

Table 2: Proper HPLC Waste Collection Options for Purchase
Safety Cans Ported (no threads) Cap 4 Port (threaded) Cap Solvent Bottle Cap
Justrite Mfg. Co., L.L.C. examples of HPLC safety cans that can be used for waste collection. VICI Jour HPLC Eco Cap VICI Jour 4 Port HPLCWaste Cap  KX VaporrSafe Solvent Safety Bottle Cap

 

Table 3: Proper HPLC Waste Collection with Modified Caps
Exhaust Filtered 2 Line Carboy 2 Line Glass 4 L Exhaust Filtered
Four liter brown glass HPLC waste container with modified cap and exhaust filter. Twenty liter carboy with cap modified for HPLC waste collection. Four liter brown glass HPLC waste collection container with properly modified cap with 2 hoses. Four liter brown glass HPLC waste collection container with properly modified cap with filter and hose.

HPLC Accessory Vendors

A container and/or cap designed for HPLC waste collection can be purchased by the research lab. There are several companies that provide HPLC waste accessories for purchase, a few are listed below:

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