Simplified determination of serum cholesterol sulfate by gas-liquid chromatography combined with cyclohexylsilane-bonded phase column purification

Abstract

A new gas-liquid chromatographic (GLC) determination of cholesterol sulfate (CS) and dehydroepiandrosterone sulfate (DHEAS) for a biochemical diagnosis of recessive X-linked ichthyosis (RXLI) is described. Although the GLC method for determination of CS is known to be more sensitive than the thin layer chromatographic (TLC) method, the former method has not been widely employed because of its complicated pre-purification steps. The present method allows us to measure the serum levels of CS and DHEAS without tedious purification steps such as multiple conventional column chromatography and preoperative thin layer chromatography. Sulfated steroids are rapidly purified with a commercially available mini disposable cyclohexylsilane-bonded phase (CH) column, CH BOND ELUT, and the purified steroids after desulfation are converted to water-resistant tert-butyldimethylsilyl ether derivative for the GLC analysis on dual 2m glass columns packed with 2% XE-60 on Chromosorb W. By the present method, serum CS concentrations in RXLI patients were shown to be about 10 times higher than those in patients with ichthyosis vulgaris, carriers of RXLI, and healthy subjects. This method is more suitable not only for a biochemical diagnosis of RXLI but also for studies on the metabolism of sulfated steroids than the previous time-consuming GLC methods.

Cholesterol sulfate in human physiology: what's it all about?

“…As noted, cholesterol sulfate and DHEA sulfate are two of the most abundant steroid sulfates in the human circulation where their concentrations overlap, although the mean level of the latter sulfoconjugate tends to be somewhat higher (34,(37)(38)(39). There is, however, a remarkable age-dependent decrease in plasma DHEA sulfate from its postpubertal peak (46,47), whereas there appears to be no age-dependent change in the level of plasma cholesterol sulfate (37).…”

“…The apparent validity of this initial value was soon confirmed by reports of plasma cholesterol sulfate levels involving a limited number of subjects that ranged from 174 to 328 g/100 ml (31)(32)(33). Studies involving larger groups of subjects but utilizing different methodologies revealed plasma cholesterol sulfate levels in normal subjects of 150-250 g/ml (34,35), 178 Ϯ 31 g/100 ml (men and women) (36), 134-322 g/ml (men) and 117-214 g/ ml (women) (37), 118-138 g/ml (men) (38), 272 Ϯ 64 g/ml (men) and 175 Ϯ 70 g/ml (women) (39), 134 Ϯ 29 g/ml (men) (40), and 253 Ϯ 25 g/ml (women) (41). Plasma cholesterol sulfate, which is carried in part by LDLs (35,42,43), can be significantly elevated in certain pathologic conditions, such as cirrhosis of the liver (41), hypercholesterolemia (36,41), and hypothyroidism (44).…”

“…Plasma cholesterol sulfate, which is carried in part by LDLs (35,42,43), can be significantly elevated in certain pathologic conditions, such as cirrhosis of the liver (41), hypercholesterolemia (36,41), and hypothyroidism (44). In recessive X-linked ichthyosis (but not other forms of ichthyosis), a skin disorder characterized by excessive scaling, the levels of plasma cholesterol sulfate are strikingly elevated, often by more than an order of magnitude (34,35,39); furthermore, the high plasma levels are associated with elevated levels of cholesterol sulfate in the stratum corneum of the epidermis (45). On the other hand, plasma dehydroepiandrosterone (DHEA) sulfate, which in young adults is normally as abundant in the circulation as cholesterol sulfate, is not elevated in recessive X-linked ichthyosis (34,35,38,39).…”

“…In recessive X-linked ichthyosis (but not other forms of ichthyosis), a skin disorder characterized by excessive scaling, the levels of plasma cholesterol sulfate are strikingly elevated, often by more than an order of magnitude (34,35,39); furthermore, the high plasma levels are associated with elevated levels of cholesterol sulfate in the stratum corneum of the epidermis (45). On the other hand, plasma dehydroepiandrosterone (DHEA) sulfate, which in young adults is normally as abundant in the circulation as cholesterol sulfate, is not elevated in recessive X-linked ichthyosis (34,35,38,39). The latter finding clearly suggests that the primary metabolic pathways for these two prominent circulating steroid sulfoconjugates are dissimilar (34).…”

Expression of Cholesterol Sulfotransferase (SULT2B1b) in Human Platelets

“…holesterol sulfate is quantitatively the most important known sterol sulfate in the human circulation, where its concentration ranges from 2 to 6 mol/L [1][2][3][4][5] ; furthermore, it is widely distributed in human tissues 6,7 and is a normal constituent of red blood cells 1,4,8 -10 and platelets. 11,12 Interestingly, cholesterol sulfate has emerged as a multifaceted regulatory molecule.…”