diff --git a/lib/galaxy/files/sources/__init__.py b/lib/galaxy/files/sources/__init__.py
index 78298024a94..455339e1043 100644
--- a/lib/galaxy/files/sources/__init__.py
+++ b/lib/galaxy/files/sources/__init__.py
@@ -3,7 +3,10 @@ import os
 import time
 from typing import Set
 
-from galaxy.exceptions import ItemAccessibilityException
+from galaxy.exceptions import (
+    ConfigurationError,
+    ItemAccessibilityException,
+)
 from galaxy.util.bool_expressions import (
     BooleanExpressionEvaluator,
     TokenContainedEvaluator,
@@ -98,6 +101,7 @@ class BaseFilesSource(FilesSource):
         self.writable = kwd.pop("writable", DEFAULT_WRITABLE)
         self.requires_roles = kwd.pop("requires_roles", None)
         self.requires_groups = kwd.pop("requires_groups", None)
+        self._validate_security_rules()
         # If coming from to_dict, strip API helper values
         kwd.pop("uri_root", None)
         kwd.pop("type", None)
@@ -191,6 +195,18 @@ class BaseFilesSource(FilesSource):
         evaluator = BooleanExpressionEvaluator(token_evaluator)
         return evaluator.evaluate_expression(rule_expression)
 
+    def _validate_security_rules(self) -> None:
+        """Checks if the security rules defined in the plugin configuration are valid boolean expressions or raises
+        a ConfigurationError exception otherwise."""
+
+        def _get_error_msg_for(rule_name: str) -> str:
+            return f"Invalid boolean expression for '{rule_name}' in {self.label} file source plugin configuration."
+
+        if self.requires_roles and not BooleanExpressionEvaluator.is_valid_expression(self.requires_roles):
+            raise ConfigurationError(_get_error_msg_for("requires_roles"))
+        if self.requires_groups and not BooleanExpressionEvaluator.is_valid_expression(self.requires_groups):
+            raise ConfigurationError(_get_error_msg_for("requires_groups"))
+
 
 def uri_join(*args):
     # url_join doesn't work with non-standard scheme
diff --git a/lib/galaxy/util/bool_expressions.py b/lib/galaxy/util/bool_expressions.py
index 5e0825d6442..cd93cb2ec89 100644
--- a/lib/galaxy/util/bool_expressions.py
+++ b/lib/galaxy/util/bool_expressions.py
@@ -150,6 +150,17 @@ class BooleanExpressionEvaluator:
             log.error(f'BooleanExpressionEvaluator unable to evaluate expression => {expr}', exc_info=e)
             raise e
 
+    @classmethod
+    def is_valid_expression(cls, expr: str) -> bool:
+        """Tries to evaluate the given boolean expression and returns True if it is valid or
+        False if it has syntax or gramatical errors."""
+        try:
+            evaluator = BooleanExpressionEvaluator(ValidationOnlyTokenEvaluator())
+            evaluator.evaluate_expression(expr)
+            return True
+        except ParseException:
+            return False
+
 
 class TokenContainedEvaluator(TokenEvaluator):
     """Implements the TokenEvaluator interface to determine if a token is contained
@@ -165,3 +176,12 @@ class TokenContainedEvaluator(TokenEvaluator):
 
     def evaluate(self, token: str) -> bool:
         return token in self.tokens
+
+
+class ValidationOnlyTokenEvaluator(TokenEvaluator):
+    """Simple TokenEvaluator that always evaluates to True for valid tokens.
+
+    This is only useful for validation purposes, do NOT use it for real expression evaluations."""
+
+    def evaluate(self, token: str) -> bool:
+        return True
diff --git a/test/unit/util/test_bool_expressions.py b/test/unit/util/test_bool_expressions.py
index caf079d7467..cf38fa0cc97 100644
--- a/test/unit/util/test_bool_expressions.py
+++ b/test/unit/util/test_bool_expressions.py
@@ -14,6 +14,27 @@ TOKEN_FORMAT = DEFAULT_TOKEN_FORMAT
 # of the *valid tokens* (those that match the TOKEN_FORMAT) will be evaluated to False.
 TOKENS_THAT_ARE_TRUE = {"T1", "token_2"}
 
+VALID_EXPRESSIONS_TESTS = [
+    ("T1", True),
+    ("token_2", True),
+    ("T3", False),
+    ("valid_token", False),
+    ("not T3", True),
+    ("NOT token_2", False),
+    ("T1 and not T3", True),
+    ("NOT T1 AND token_2", False),
+    ("not T3 or (T3 AND token_2)", True),
+    ("T1 and (T3 OR token_2)", True),
+    ("(T3 OR T1) and not (T3 OR valid_token)", True),
+]
+
+INVALID_EXPRESSIONS_TESTS = [
+    "",
+    "23 45",
+    "'some quoted str' and not T1",
+    "invalid expression",
+]
+
 
 @pytest.fixture(scope='module')
 def contained_evaluator() -> BooleanExpressionEvaluator:
@@ -26,30 +47,25 @@ def contained_evaluator() -> BooleanExpressionEvaluator:
     return evaluator
 
 
-@pytest.mark.parametrize('expr, expected', [
-    ("T1", True),
-    ("token_2", True),
-    ("T3", False),
-    ("valid_token", False),
-    ("not T3", True),
-    ("NOT token_2", False),
-    ("T1 and not T3", True),
-    ("NOT T1 AND token_2", False),
-    ("not T3 or (T3 AND token_2)", True),
-    ("T1 and (T3 OR token_2)", True),
-    ("(T3 OR T1) and not (T3 OR valid_token)", True),
-])
+@pytest.mark.parametrize('expr, expected', VALID_EXPRESSIONS_TESTS)
 def test_expression_evaluates_as_expected(expr: str, expected: bool, contained_evaluator: BooleanExpressionEvaluator):
     actual = contained_evaluator.evaluate_expression(expr)
     assert actual == expected
 
 
-@pytest.mark.parametrize('expr', [
-    "",
-    "23 45",
-    "'some quoted str' and not T1",
-    "invalid expression",
-])
+@pytest.mark.parametrize('expr', INVALID_EXPRESSIONS_TESTS)
 def test_invalid_expression_raises_exception(expr: str, contained_evaluator: BooleanExpressionEvaluator):
     with pytest.raises(Exception):
         contained_evaluator.evaluate_expression(expr)
+
+
+@pytest.mark.parametrize('expr, _', VALID_EXPRESSIONS_TESTS)
+def test_is_valid_expression_return_true_when_valid(expr: str, _: bool):
+    result = BooleanExpressionEvaluator.is_valid_expression(expr)
+    assert result is True
+
+
+@pytest.mark.parametrize('expr', INVALID_EXPRESSIONS_TESTS)
+def test_is_valid_expression_return_false_when_invalid(expr: str):
+    result = BooleanExpressionEvaluator.is_valid_expression(expr)
+    assert result is False