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Endotoxin Detection Reagents Limulus PS Single Test Wako

  • Turbidimetric Technique
  • Endotoxin Specific

Features

  • Endotoxin Specific measurement can be performed with small amounts, avoiding any interference from coexisting substances, by adsorbing endotoxin to Pyrosep™ in the sample and condensing it.
  • Since the endotoxin is adsorbed to Pyrosep™ even in ethanol, a fat-soluble sample can be tested provided it dissolves in ethanol. Such samples are not usually processed in ordinary limulus tests. (Example: fat-soluble vitamins, oils and fats)
  • No LAL reagent is wasted, since it is pre-dispensed individually in vials, and can be used in the quantity required for measurement.

Measurement Principle

The endotoxin in the sample is adsorbed to a capillary column filled with Pyrosep™ (an affinity adsorbent with histidine bound to a water-insoluble support via a spacer, designed to specifically adsorb endotoxin). Other substances contained in the sample are washed off, allowing the adsorbed endotoxin activate the LAL reagent for quantification, whereupon the measurement is performed using a Toxinometer®.

Measurement Principle

Kit Contents

(1) LAL ES Single reagent (lyophilized Limulus polyphemus amebocyte extract)20 vials (for 0.3 mL)
(2) Pyrosep™ suspension (suspended in a phosphate buffer)20 vials (0.77 mL)
(3) LAL diluent2 vials (7 mL)
(4) Wash solution4 vials (11 mL)
(5) Sample dilution buffer4 vials (10 mL)
(6) Capillary (with a silicone tube)20 pcs

• The syringe, T-shaped stopcock and dedicated adapter are not included in this kit, but are optionally available in the PS Accessory Kit (294-33311).

Pyrosep™ Schematic Diagram

Measurement Method

Application

A. Endotoxin Detection in Dialysate

The samples were prepared from commercially-available dialysate (dialysate X) and two kinds of replacement fluids for artificial kidneys (fluids Y and Z), with amounts of 5.0 mL in each case respectively. Each had USP Reference Standard Endotoxin added, and the pH was subsequently adjusted to approx. 5 using acetic acid. The endotoxin concentration in each sample was measured using the Pyrosep™ method and the added endotoxin was recovered.
With the use of the Pyrosep™ method, endotoxin in dialysate is detectable within a range starting from 0.1 EU/L.

Standard curve
(sample volume: 5.0 mL)
No. Specified Concentration (EU/L) Tg(min) Average Tg (min)
1 1.000 53.0
53.4
54.4
53.60
2 2.000 41.6
41.2
41.18
41.53
3 10.00 26.6
26.0
25.6
26.07
4 100.0 14.6
14.8
14.4
14.60
Table 1: Endotoxin Recovery from Peritoneal Dialysate/Replacement Fluid Using the Pyrosep™ Method
SampleEndotoxin
Spike (EU/L)
Endotoxin
Recovery (%)
Dialysate X 0.1
1.0
80
72
Fluid Y 0.1
1.0
144
102
Fluid Z 0.1
1.0
108
118

B.Endotoxin Detection in Fat-soluble Vitamin

Fat-soluble vitamins D, E and K were dissolved in ethanol, to which USP Reference Standard Endotoxin was then added. The endotoxin concentrations in the samples were measured using the Pyrosep™ method and the added endotoxin was recovered from each sample. The use of the Pyrosep™ method enabled the endotoxin detection in fat-soluble vitamins, which could not otherwise be measured with Limulus reagents.

Table 2: Endotoxin Recovery from Fat-soluble
Vitamins Dissolved in Ethanol Using the Pyrosep™ Method
VitaminConcentration
(mg/mL)
Endotoxin
Recovery (%)
D2 10
170
E 10
20
91
83
K 10
20
99
68

・Endotoxin addition amount/concentration adjusted to make the final concentration 0.05 EU/mL.

C.Other Examples

Ethanol, silicone oil, soybean oil, olive oil, stearic acid, egg yolk lecithin, oil-adjuvanted vaccine, albumin products, coagulation factor products (factor 8), interferon formulation, antithrombin Ⅲ, immunoglobulin product and other ethanol-soluble samples

Single test Type
Code No. Product Name Quantity
299-54501 Limulus PS Single Test Wako 20 tests
Accessory Kit
Code No. Product Name Quantity
294-33311 PS Accessory Kit 1 set

Kit contents:
20 mL syringe (no needle) x 12, T-shape stopcock R type x 12, dedicated adapter x 20, bulldog clip x 12, 352058 FALCON polystyrene tube (5 mL) x 25, 352057 FALCON polystyrene tube (14 mL) x 25, test tube rack (for 50 tubes) x 1 and procedure instruction document x 1

Topics

  • LAL Basic Knowledge
  • Lysate reagent selection flowchart

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